loHlnn Untttrrattg QlnlUgp nf Hib^ral ArtB

The Gift of . ..ir.ke TT^utWor.

BOSTON UNIVERSITY GRADUATE SCHOOL

Dissertation

NEURO-MUSCULAR ACTIVITY IN THE PEDAL WAVES OP HELIX

by

Blanche Brine Daly (A.B,, Hunter College, 1913; M.Sc, Nev/ York University, 1915; M.A., Radcliffe College, 1928) submitted in partial fulfilment of the requirements for the degree of Doctor of Philosophy 1935

4q ^ 11

-PhD 19 33

ACKNOWLEDGMENT The writer wishes to express her

«

appreciation to Professor Brenton h. Lutz , Department of Biology, Boston University, for his helpful advice and continued interest in this work.

CONTENTS

CHAPTER I Introduction Page i

CHAPTER II Purpose of this investigation 5

CHAPTER III Review of the literature 10

CHAPTER IV Method and procedure 19

CHAPTER V Results and discussion 26

A. Creeping during vertical

ascension without loads 26

B. Comparison of vertical creeping

with and without loads 33

G. The effect on locomotion of

de-eying Helix 43

Behavior of the detached foot 49

E» Effect of mechanical stimulation

and of Ringer's solution on locomotion 51

F» Action of adrenalin on the

neuro-muscular activities

during locomotion 62

Effect of strychnine sulphate

on locomotion 98

CHAPTER VI Summary and conclusions 115

CITATIONS 118

INDEX OF FIGURES

123

1

Chapter I. Introduction

Precise measurement of vital processes is necessary in the investigation of many of the problems arising in the study of vital dynamics. Results based on series of facts obtained upon the intact living organism prevent confusions which arise when less exact methods are employed. The philosophical approach into the realm of vital phenomena has not been a step forward because the introduction of philosophical ideas, such as adaptation, behavior, emergence, psychobiology and purpose have resulted only in methods of vagueness and inexactness.

Huxley (1854) predicted that the science of the biologist would be as deductive and exact as mathematics. He was of the opinion that since biology is a physical science the methods of this science must be analogous to those followed in other physical sciences. This prediction was fulfilled sooner than he would have believed possible. Loeb (1888) laid the foundations for this method of precise measurement by his contribution of tropistic conduct. Crozier (1928) has taken tropisras as a working tool for ttie further devel- opment of many unexplained activities found in living phen- omena. He recognizes the significance of the quantitative aspects of behavior and believes that it is this quantita- tive treatment of tropistic behavior which is essential if an understanding of conduct is to be furthered, ^.s he has

I

2

stated, the biological system presented by a sin^^e individ- ual is not a "thing", a single event, but a system of rela- tions. These relations must be defined through investigatio! and their functional dependence analyzed.

Stier (1928) has mentioned certain aspects of animal conduct other than those considered in the tropism doctrine; that is, motor activity and fluctuations of sensitivity may be based on principles originating in the fundamental laws of chemistry and physics.

Rose (1929) emphasized the fact that it is only by more precise measurement that the problem of vital dynamics can be cleared up, and its progress can be furthered only by the use of quantitative methods and rigorous analyses. By delicate measurements scientists have obtained just such desired results, which have completely revolutionized researches in the field of plant tropisms and at the same time have opened up new aspects of the physiology of plants.

Furthermore, various related phenomena of animal reac- tions cover a wide field. Myogenesis (Carey, 1919-20), oxidations as a function of temperature (Crozier, 1924-25), tonic immobility (Hoagland, 1927), spontaneous movement (Stier, 1928) and inheritance (Crozier and Pincus, 1929-30) are a few activities taken from the field of animal conduct that have led to important results by the application of the method of precise measurement. For example, Crozier and Pincus have arrived at a definition of gene differing from the usual one; for them it is a definition of the

3

effect in inheritance as a function of some controlling, independent variable.

In the investigation of the dynamics of histogenesis, Carey (1919-20) has shown by the method of precise measure- ments (the number of contractions made in a unit of time, measurement of intra-vesicular hydro-dynamic pressure and volume of stimulus) that the formation of muscular tissue is due to a definite active process, not a passive one, as the term self-determination denotes. He proved by his experiments that the intensional stimulus is the necessary factor in myogenesis. From his dynamic point of view the muscle types represent differences in the amount of work that has been done on the undifferentiated mesenchyme by the differentially growing parts of the embryo during the active growth period. He demonstrated that it is possible to transform unstriated muscles into striated by varying the velocity of application and the intensity of the ten- sional stimulus to a higher optimum degree. 3y this taethod of procedure his results enabled him to conclude that the variable intensity of the optimum tension determined the muscular type. In other words, the structure of muscular tissue is determined by the function it performs and the work it does, but on the other hand structure does not deter- mine function. He thus reached these conclusions as a result of his experiments employing analysis through tension and pressure.

Therefore, just as analysis through temperature

characteristics plays an important part in the snocific control of vital processes, so also analysis tiiroufih pres- sure and tension becomes an additional factor which may lead to identification of reacting living matter. The fol lowing experiments are approached through this procedure.

5

Chapter II. Purpose of this investigation

The purpose of this investigation is the detailed analysis of the factors involved in locomotion, to eain additional information concerning the creeping mechanism of Helix. Inasmuch as the movement of the waves found in the foot of gastropods is inseparable from locomotion it is necessary to describe the types of waves found in these animals; and it is also of importance to consider other forms of periodic waves found in muscular tissue.

Various kinds of periodic waves occurring in muscular tissue have been investigated to determine, if possible, the cause of neuro-muscular activity. The waves which are present in the peristaltic action of the intestine, the rhythmic pulsation of the cloaca in holothurians , the pedal waves found in larvae of slug-moths, the waves found in Thy one briareus , and the peristaltic locomotor waves of the tent caterpillar may be cited as examples.

The balance of opinion as to the cause of the rhythmic contractions of the intestine has been in favor of neuro- genesis, but there is now considerable evidence of tayogenic origin, offered by Ivlagnus (1905), Gunn and Underhill (1914), Alvarez and Mahoney (1922), Gowie, Parsons and Lashmet (1929), Ascanio and ..Ivarez (1929). Alvarez agreed with some of these investigators as to myogenic origin of peristaltic action and believed that a gradient of irritability has been

6

found which may be an Important factor in the directlor normal peristalsis.

The anal pulsating mechanism of holothurians might be regarded as constituting another unit in the series of inde- pendent effectors, such as pedicellariae and spines, which go to make up the echinoderra meuro-muscular equipment. Crozier (1916) has investigated the physiological character- istics of cloacal pulsations in Stichopus moebii Semper. The results of these experiments are in essential agreement with the data derived from many previous studies in pulsating structures, such as those of medusae, ctenophores, the arthropod heart, and the vertebrate heart and intestine,

e» J the rhythm has a temperature coefficient of the order of magnitude of that for chemical processes and the relation of pulsation to the salts of sea water is essentially like that in other well-known pulsating systems.

The pedal waves found in the larvae of the slug-moths (Gochlidiidae) are similar in many respects to those on the pedal surface of gastropods. Crozier (1923-24) has found that the speed of these pedal waves corresponds almost exactly with the speed of the pedal wave in Chiton tuberculatus and is therefore in this respect nore comparable to the raol- luscan foot than to the peristalsis of the body in the earth- worm or in caterpillars. He has concluded that the peris- taltic pedal waves of these animals are to be regarded as "a derivative of the general peristalsis of ordinary caterpillars, and as in the 'myenteric reflex' of tlie vertebrate intestine

7

it implies reciprocal innervation." (p.. 328)

When the peristaltic locomotor waves of the tent cater- pillar were studied by Crozier and Stier (1925-26) it was found that the frequency of the abdominal waves during ver- tical ascension was controlled by temperature according to the Arrhenius equation.

Stier (1928) has come to the conclusion that a proprio- ceptive mechanism in the body wall seems to control the initiation of the locomotor waves in Thyone.

Lutz (1930) studied the effect of low oxygen tension on the pulsations of the isolated holothurian cloaca (isolated strips from the cloaca of S-tichopus moebii Semper and ring preparations from the cloaca of Gucuinaria frondosa ) because oxygen deficiency has often been associated with periodicity and augmentation of response in various tissues. He concluded that a certain degree of oxygen lack results in increased activity of the tissue.

Although the exact cause of the initiation of the rhythmical waves (referred to in the preceding paragraphs) is not known, it can be seen that by means of investigations of this nature new factors are brought to light that play an important part in the control of periodic waves.

To investigate rhythmical waves further the gastropods offer excellent opportunities, and the same principle when found may apply to other tissues where periodic waves occur. The action of the waves found in the foot of gastropods has frequently been compared to the peristaltic waves occurring

in the stomach and intestine. Jordan (1927) found that the cerebral ganglia of snails have a quantitative influence on the peristaltic action of the foot (v/ith its pedal r^an-? i similar to the autonomic innervation of the stomacn. how- ever, the use of the foot of this animal is of advantage compared with the use of other tissues such as excised pieces of intestine. In the study of the latter many more factors enter, such as regulation of temperature and of intra-intestinal pressure, and the use of oxygenated Locke's solution, as well as other factors, thus adding to the com- plication of investigation.

Helix pomatia (Pig. Al, a and b) and Helix lactea (Pig. Al, c and d), common Liediterranean species, were used in these experiments, which extended over a period of three years. As the animals possess great tenacity of life and are unaffected by extremes in temperature they are adapted to experiments continuing over long periods of time. Another advantage is their precise negative geotropism. In the experiments of vertical creeping v/ith and without loads this tropistic quality insures upward creeping. Still another advantage of using Helix is the tjj)e of wave it possesses, spreading over the entire foot. There- fore, the adhesive power of the foot does not present such a- complication as in the foot that does not form pedal waves over its whole breadth. Helix is moreover very suitable for analysis through precise measurements on the

9

intact organism.

Just as investigation through temperature character- istics has thrown light on the specific control of vital processes, so analysis through pressure and. tension becomes an additional factor that may lead to the identification of reactions in living matter. As the central nervous interplay governing the mechanism of locomotion and the exact cause of the waves is unknown in gastropods, the subsequent experiments v/ere undertaken to investigate further these waves and other factors involved during locomotion under varying conditions. The quantitative measurements of the activities investigated in these animals have been studied and their functional dependence analyzed.

1

Crozier and Pilz (1923-24) have stated that it should be possible to utilize these waves for the study of neuro- muscular physiology in the intact animals, avoiding in this way effects due to lack of proper circulation and the like when isolated organs are used.

10

Chapter III. Review of the literature

The pedal waves found, in the foot of the gastropods have been described and in some cases classified by Carlson (1904-05), Biederraan (1905, 1906), Vies (1907), Parker (1911, 1914), Olmstead (1917-18), van Riynberk (1918-19), Crozier (1918-19); 1919-20; 1922-23 a and b) , and ten Cate (1923). Tlie creeping of Limax maximus has been described by these writers as a result of a rhyth^nic succession of evenly spaced, progressive wave-like deformations of the pedal surface.

Vies (1907) has grouped the v/aves that appear on the foot of mollusks into two general types, i.e., the "direct" waves that pass from the posterior to the anterior end of the foot when the animal moves forward, and the "retrograde" waves v/hich pass over the foot from front to back as the animal m.oves forward."^ In both types of movement several subtypes can be distinguished as determined by the lateral extent of the pedal wave, i.e., "monotaxic", "ditaxic" and "tetrataxic" subtypes. The direct waves may be m.onotaxic, ditaxic and tetrataxic types, the retrograde waves either

With the exception of Chiton gastropods always i.iove forward and never backwards, regardless of the type of waves passing over the foot. Parker (1914) has found that Chiton can reverse its locomotion and creep backwards a few millimeters. This is the first instance of backward loco- motion to be recorded in gastropods (verified later by Crozier and Navez).

11

monotaxic or ditaxic. The direct monotaxic waves consist of a single system of waves traversing the foot as found in Limax and Helix. Direct ditaxic waves consist of two systems of waves occupying each one of the lateral halves of the foot and alternating regularly on the two sides of the median line which is not affected by these waves. Parker (1911) has added Tectarius nodulosus and Nerita tessellata as ditaxic gastropods and has found that in these animals the waves on the two sides of the foot usually alternate and are so extensive that never more than two waves can exist on one side at one time. As a result of this the foot moves forward in alternate steps, first on the right and then on the left, the motion resembling that of a person walking in a sack. Vies (1907) has described the direct tetrataxic waves, i.e., the foot which is traversed by four systems of waves is broadly fissured on the median line and each lateral sole is overrun by two systems of alternate waves such as are found in the ditaxic foot. The retrograde waves may be monotaxic or ditaxic. In considering the theories advanced to explain the locomo- tion of gastropods only the first group (direct monotaxic) is considered by the following authors.

The investigation of the activity of these waves has resulted in various conclusions concerning both the mechan- ism controlling them and other factors in locomotion.

In Helix the waves are m.onotaxic, but they pass over the entire breadth of the foot (helicine foot), whereas in Limax they appear only in the central longitudinal area of the foot.

12

Contraction of the longitudinal muscles, the action of the dorso-ventral muscles (transverse and oblique rauscles), the pressure of substances in the body fluids, and other factors have been advanced to explain the controlling mechanism.

Simroth (1878) accounted for locomotion by a theory that he called "extensile muskulatur". He concluded that the waves cannot be produced by the separate or the combined contractions of the oblique and transverse muscular strands, but that the cause of the extension of the foot is the active extension of the longitudinal musculature. He believed that the extensile muscle fibers in all or nearly all snails are the important factors in its m.ovement from place to place.

Jordan (1901) advanced a theory which he later r.iodi- fied. He did not accept the theory of "extensile muslrulatur" in accounting for locomotion in the m^^rine gastropod .^tplysia. He attributed the relaxation or the extension of the longi- tudinal muscle of the foot to pressure of isolated bodies of the visceral fluid or blood.

Bohn (1902) attributed the production of the waves to direct excitation of the m.uscle fibers and to a sort of progressive "induction". He explained that this "induction" would be produced every time the m\iscle fibers v/ere placed on level cylindrical or conical surfaces and the waves would be distributed in bands or rinfi;s, parallel, narrow and identical vath each other. He called then organic waves and postulated the theory that by the study of these

13

and other orp;?:nic waves other facts of biolop-io i^i^I^'ctlon may be brought to li,(7,ht and may play an important role in an explanation of the kinetogenetics of evolution and per- haps even of heredity.

Carlson (1904-05) has described the external mechanics of locomotion and has differed in some respects from other authorities. He did not accept Simroth's theory of "ex- tensile muskulatur" or Jordan's theory of the pressure of isolated bodies in the visceral fluid. Carlson tested the longitudinal muscle in the foot of several gastropods and his results showed that there is no difference between the physiology of the muscle and that of any other muscle. He concluded that during ordinary progression the animal assumes its greatest length and smallest diameter, due to the contraction of the transverse and the oblique muscles of the dorsal and lateral parts of the body. The waves of locomotion in the foot are diminutive representations of the waves of relaxation and contraction. At the areas of relaxation the sole of the foot adheres closely to the ground and between these points the sole is slightly elevated. Although von Uexkllll believed that the foot is provided v/ith some mechanical device such as the setae of the earthworm, Carlson believed that the area of contact of the foot with the ground in any region serves as a fixed point through friction, and acting on this the contraction of the longitudinal muscles of the foot pulls the neighbor- ing portion of the body forward.

14

Parker (1911) has described most satisfactorily the external mechanics of locomotion in Helix pomatia. fie stated that the forv/ard movement takes place in the dark waves, and quiescence is characteristic of the intermediate lighter portions of the foot. Each wave is a pulse of for- ward motion and the rest of the foot is momentarily quies- cent. He stated, "the area covered by the v/ave is probably a fourth or fifth of the wnole foot, any moment, there-

fore, three-fourths to four-fifths of the surface of the foot is stationary and about one-fourth to one-fifth is moving forward. In other v/ords, the snail stands on the greater part of its foot while it moves- forward with a much lesser part." (p. 102). Prom, experiments performed he believed that each wave on the underside of the foot (of flelix pomatia ) is a slight concavity. I'Vhen the muscle of the foot relaxes the portion of that foot that v/as elevated is returned to its former level and the muscle recovers its original length and position. This action of the dorso-ventral muscles takes place from behind forward and thus a concave wave runs on the surface of the foot from tail to head. The forv/ard movement of that portion of the foot which is temporarily lifted from the substrate is accomplished by the action of the longitudinal muscles. The contraction of each longitudinal fiber serves to move the foot forward as the relaxing wave passes over the foot. It also extends the relaxing posterior fibers. An important point in this description is that the dorso-ventral :nuscles

15

play an active part in lifting the foot from the substrate due to the fact that their dorsal ends, being more firmly set than the ventral ends, serve as relatively fixed points and therefore the ventral ends move. The action of the ventral' end lifts the foot locally and overcomes adhesion in the given region. In this way each point of the foot is lifted, moved forward and set down again and thus the foot, and with it the anim.al as a whole, moves forward.

Although Vies and Bathellier (1920) did not arrive at a conclusion in regard to the action of the pedal waves of gastropods they deduced that certain numerical laws held for them.

ten Gate (1923) was of the opinion that the wave of contraction is caused by the nervous peripheral net-work.

Grozier and Pilz (1923-24) agreed with the description of the external mechanics of locomotion as given by Parker (1911). It appears from Grozier 's investigation of the neuro-muscular activity of the foot that he did not have at first conclusive evidence of nerve-net transmission. However, from the experiments (Grozier and Pilz, 1923-24) performed on the temperature coefficient for pedal activity in Limax these investigators found that the velocity of a single wave must have very nearly the same temperature characteristic which is found also in another case of nerve-net transmission (Renilla ) . They found that v/hen work was done at a constant rate the frequency of the pedal waves is influenced by the temperature according to the Arrhenius equation, with ^ = 10,700 (Qio 11° to 21°G. = 2.1).

16

Magnus (1924) has shown from the "half animal" experi- ments performed by Jordan and Uexkflll the.t stretchinr-r of the musculature in snails can react on the condition of the pedal ffanglia.

Crozier and Federighi (1924-25£) concluded that the pedal organ of the slug, although under the control of cen- tral nervous impulses, is essentially an independent effec- tor. (See further, page49 of this thesis.)

Cole (1925-26) considered the stimulus for geotropic orientation and locomotion in Helix aspera to be the tension of the body muscles produced by the downward pull of gravity The stim.ulus is received by the proprioceptors of these muscles .

Jordan (1927) explained the rhythmical transmissions of stimulation in the snail's foot according to von UexkiJlll' law (p,243). The foot of the snail, with its nerve net, is not sufficiently autonomous to be able to carry on creeping without the help of a ganglion. The stimulation which pro- duces locomotion is conducted in the beginning through the nerve-net and through the pedal ganglia with the nerves radiating from them to the periphery, as to the manner in which the m.ovement itself comes into existence, nothing exact is known. A true antagonism such as appears in arthro pods, annelids and vertebrates is lacking in these animals, and instead the phenomenon of "viscosoid" tonus is present.

Jordan considered "viscosoid" tonus as a static phenomenon and as an attribute of a colloid system. In his

I

17

distension experiments on the snail's foot he stated that the characteristics of these muscles can be reco(=i;nized as those of a fluid colloidal phase. The rausculature of these animals possesses attributes which in many connections have the characteristics of protoplasm. For instance, piiago- cytosis is present in considerable degree in the same animal in which "viscosoid" tonus plays so important a role in the outer muscle layer. In amoeboid movements and phagocytosis the work is connected with the shifting of the parts and thence with an alteration of the form. IVith regard to muscle, therefore, a new form of contraction problem arises. As soon as the musculature is shortened • through stimulation nothing more is observed of the fluid phenomenon. The con- tracted muscle is an almost pure elastic body. In tnis con- nection Freundlich, as referred to by Jordan, has stated that in dead colloidal stuffs the conversion of the fluid- viscoid condition into the elastic state occurs. Hypotheti- cally one may think, in the case of the muscle of a conver- sion from the sol to the gel condition. In the latter the form-alteration would no longer be explicable through the shifting of the component parts bu.t through a particular reversible elastic form-alteration of them.

ten Gate (1927-28) has stated that the movement of the appendages of Aplysia limacina is determined directly by the pedal ganglia and is not Produced by a chain of ganglia.

As real geotropic orientation of the Helix type of gastropod takes place only during active prop-ression.

18

orientation, because of its close relationship to locomo- tion, can "be included in this discussion,"^

(jeotropi'c orientation rather than locomotion has been studied by Grozier and Kavez (1930). They concluded that orientation is not governed either as to direction or as to latent period by the involvement of the statocyst. Gravitationally excited orientation is probably controlled by the proprioceptive stimulation through impressed tensions (Grozier and Navez, 1930). As orientation and progression are so closely united these investigators have stated that it is still a question as to whether (1) the creeping move- ment and postural movement are released together or (2) the operation of the pedal wave is the necessary factor to act on the stimulus for orientation. Therefore it can be seen from various theories existing v/ith regard to this subject that it is important to investigate the question further. Helix, rather than Limax was chosen, as more work has be-en done on Limax (Grozier and Pilz, 1923-24; Grozier and Federighi, 1924-25£) and very few data have been ob- tained for Helix.

The muscular mechanism for locomotion and that for orientation is structurally distinct, i.e., creeping is brought about by means of a pedal organ, wxiile orientation is accomplished by means of muscles in the dorso-lateral regions of the body wall.

19

Chapter IV

Method and Procedure

Specimens of Helix lactea and Helix pomatia used in these experiments were kept at room temperature in separ- ate jars which were kept slightly moist. Lettuce v/as fed every other day and the jars were thoroughly cleaned. In this way the animals lived for six or seven months, the vigor of creeping showing no diminution at the end of that period.

The following experiments were carried on within the

temperature lim.its of 19.5° and 24.8°C. The animals were

subjected to varying environmental conditions, which will

be described subsequently. The required measurements were

obtained as follov/s; the animal was placed in a glass jar

1

one and one half feet high and when the snail began to creep vertically upwards, the sides of the jiar being kept moist to promote active creeping, the distance travelled while ten waves were passing over the foot from posterior to anterior end was measured in millimeters on the outside of the jar. (Pig. 1). The time was recorded in seconds or minutes. This constituted one run, and ten runs made up a series (see Table I). Several series for each animal were obtained and the conditions for each series were the

^ In later experiments a glass plate set in a frame similar to a picture frame was used, as this enabled handling the animal more easily.

20

same. The length of the foot during each run was also measured in millimeters.

Prom these data it is possible to determine:

(1) The frequency of the pedal waves (F).

(2) The velocity with which a single wave traverses the foot (v).

(5) The speed of creeping (V).

(4) The advance due to a single wave (A).

An example of the way in which the data observed have

been used to obtain the absolute values of the frequency

1

of waves and the velocity of progression is presented in detail in Table I, and a description of the way these absolute values are used in plotting one relation to another is given in the text.

As the ultimate purpose of this series of experiments was to analyze the relation between the frequency of waves and the velocity of progression it was necessary to obtain from the data thus observed the values of the velocity of progression, which are given in Table II. With these measurements it is possible to show by their actual scatter and by comparative graphs the linear relationsiiips between the different activities involved in locomotion. The method of plotting was to take the computed value of the frequency of waves and the velocity of progression (Table II)

The same method was follov/ed to obtain the measurement for the advance per wave and the velocity of a single pedal v/ave.

I

21

and plot the actual scatter, the ordinates representinp; the frequency of v/aves and the abscissae the velocity of progression. After presenting the actual scatter in this way their average points were obtained. The best fitted line v/as then drawn through these average points. The graph resulting from the data just given in detail is shovm in Pig. 3 for animal Ko. 7. Graphs shov/ing the same relationship have been com.pared in subsequent experiments when the data are derived under varying conditions.

Tables Ila and lib represent the data for a complete experiment when each one of the absolute values v/as deter- mined, i.e., velocity of progression, frequency of waves, velocity of a single pedal wave and the advance per wave. From these data the various graphs v/ere plotted as described above.

22

TABLE I

Observed data"

Calculated values of frequency of waves from observed data

No. IIo. Ko. Actual Actual Length of Frequency of

of of of time in distance foot during wavcg-^^ Y^^

runs series waves seconds

m mm.

each run^

time

1

10

16.0

7.5

49.0

0.624

2

10

10.8

7.5

51.0

0.925

3

10

13.9

7.0

51.0

0.719

4

10

13.9

8.0

52.0

0.719

5 6

10

= 1

10

12.4 11.2

8.5 7.5

54.0 53.0

0.806 0.892

7

10

12.2

10.0

53.0

0.819

8

10

12.0

7.5

54.0

0.832

9

10

11.4

8.5

53.0

0.878

10

ro

10.4

10.0

54.0

0.961

^ Anj.mal TIo. 7. 2

These measurements are needed v/hen the velocity of a

^ ^ . ^ • tr Length of foot

single wave is to be determined, i.e., V = t^^q for i wave *

23

TABLE II

Calculated values of progression 1 obtained from

Observed data observed data

No, No. No. ^LCtual j.ctual Length of Velocity of

of of of time in distance foot during .procuress

runs series waves seconds in mm. each run = di stance ("im_.J,

tin-:e ( sec . )

1

10

16.0

7.0

49.0

0.437

2

10

10.8

7.5

51.0

0.694

3

10.

13.9

7.0

51.0

0.503

4

10

13.9

8.0_

52.0

0.574

5

10

12.4

8.5

54.0

0.685

= 1

6

10

11.2

7.5

52.0

0.669

7

10

12.2

10.0

53.0

0.819

8

10

12.0

7.5

54.0

0.624

9

10

11.4

8.5

53.0

0.745

10

10

10.4

10.0

54.0

0.961

Animal No. 7

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26

Chapter V.

Results and Discussion

A. Verti.cal ascension without added load.

In a series of experiments extending over a period of several months eleven animals. Helix lactea, of vary- ing v^eights v/ere used. Measurements were obtained for a total of 36 series, each series made up of ten runs, and each run made up of ten waves; thus 3600 waves were ob- served.

Data were obtained from observing 400 waves in each of the animals when creeping vertically upv/ards carrying no load. The relations betv/een the velocity of a singl'e wave and the velocity of progression, and between the frequency of waves and the velocity of progression, as well as between the advance per wave and the velocity of pro- gression were analyzed.

Fig. 2^ shows the relation a single pedal wave and the rate It can be seen that a linear relationship exists regardless of the weight of the animal.

The actual observations were plotted taken from data obtained from 4 series of 40 runs, each run consisting of the record of the time and distance covered for 10 waves. In order to obtain the average points of Fig. 2 the mean was taken of the actual observations. This was the way in which the points were determined in the other figures. It is possible to obtain the average deviation from the mean for any of the points given in the figures.

between the velocity of of creeping ^,

i

27

Weight Slope of line"^ Animal Ko. 1 6.5 gm. 0.20 " "4 8.6 " 0.36 " " 7 7.9 " 0.34

Fig. 3 shows the relation between the freauency of

) . It can

waves and the velocity of nrogression (F

V

he seen also that a linear relation exists regardless of the weight of the animal.

Weight Slope of line Animal Ko. 1 6.5 gm. 0.20 " " 7 7.9 " 0.18 " "10 3.8 " 0.20

Pig. 4 shov/s the relation between the advance per

To find the slope of the line as given in Fig. a, the distance AB expressed in mm. is subtracted from the dis- tance CD expressed in mm. and divided by the distance represented by line AG. The same unit for AG is used in each case, i.e. , 50 mm.

28

The results

wave and. the velocity of pro/3;ression (^ )

V

show that in this relationship also a linear rele tion3xj.ip exists regardless of the weight of the animal.

Weight Slope of line Anirfial Ko. 4 8.6 gm. 0.5 " " 7 7.9 " 0.3 " " 10 5.8 " 0.3

Further experiments of this nature were performed on a number of different animsls to see whether consistent results v/ould be obtained. The following summary shows the scope of the experiments from which data were obtained for the three different relationships: V

V V

A for animals 1, 3, 4, 5, 6, 7, 8, 10 and 11

V

during vertical ascensions without load. For these nine animals a total number of 3200 waves was observed and measurements taken. From these data graphs were plotted of the actual scatter for each animal showing the relation between the velocity of a single pedal wave and the rate of progression (V j ), betv/een the frequency of waves

V

and the rate of creeping (F

) and betv/een the advance ). The

V

per wave and the velocity of progression (A

V

average points of each graph v/ere obtained and the best fitted line dravm. Fig. 4A shows a comparative graph of these lines, each line illustrating for each animal the relation between the velocity of a single wave and the

29

velocity of progression. A straight line relationship is found to exist under these conditions. The slope of the line for each animal was determined. The same method of comparison and analysis was employed in the investigation of the relation between frequency of waves and the velocity of progression (Fig. 4B), and "between the advance per wave and the velocity of progression (Fig. 4C ) . These results are given in Table III. ^

Kelix pomatia was used in a series of experiments to determine whether the laws derived from observations of Plelix lactea would be confirmed. The conditions and method of these experiments were the same as for Helix lactea. The limits of temperature v/ere 16.2 to 21.0°C., but constant for each experiment. The length of the foot of Helix pomatia vmen creeping is from 60 to 100 milli- meters (Figs. Ala and Alb), and that of Helix lactea from 25 to 40 millimeters (Figs. Ale and Aid). Helix pomatia has the helicine foot, i.e., it forms waves over the whole breadth of its foot, so that adherence and progression is not attended to by functionally separate divisions of the pedal surfaces. Table Ilia"'' shows some average results of these experiments. Fig. 4D illustrates that for Helix pomatia a linear proportionality exists between the velocity of a single wave and the rate of creeping. Fig. 4E shows that the frequency of waves is directly proportional to

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the rate of creeping. Fig. 4P demonstrates that the advance per wave is directly proportional to the rate of creeping.

Analysis of the experimental results obtained in this series yields certain information regarding the condition of the foot during vertical ascensions without load. A study has been made of the relations betv/een the velocity of a single pedal wave and the velocity of progression, between the frequency of waves and the velocity of pro- gression, and between the advance per wave and the velocity of progression. The slope of the line for these relation- ships has been determined. A comparison of these slopes shows the deviation of the linear proportionality, if any. Variations from strictest proportion found occasionally may arise in large part from the complex character of the pedal musculature. In general the linear proportionality of these relationships to each other is unmistakable. The relation between the velocity of progression and the veloclt^r of a single v/ave shows the greatest alteration during a given number of runs under the same conditions for a number of animals of varying weights. The relation between the frequency of waves to the velocity of progres- sion exhibits the least alteration within a number of series of one anima]. or when several animals are compared with each other, study of the graphs and tables shows that the slope of the line in a given relation may be the same regardless of the iveight of t-"'e rrimal.

Therefore, these results Siiov/ b-.at the speed of

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creeping for Helix lactea and Ilellx pome-. t la without load on a vertical surface during upv/ard progression is directly proportional to the velocity with which a single average wave courses over the foot. It is independent of the weight of the animal and the length of tlie foot. ITie velocity of progression is directly proportional to the frequence of waves and to the advance of a single wDve.

B. Vertical ascension v/ith added load.

The technique for adding the loads was to attach to the shell lead v/eights held on by pieces .of adhesive tape. The animals (Helix lactea ) were first tested to determine the weight necessary for complete exhaustion. This was for the purpose of gaining an indication of the probable number of weights to be attached and to select individual weights that could be added conveniently. Table IV represents the results when the animals v/ere tested for complete exhaustion.

TABLE IV

No. of Animal Complete Exhaustion Weight of Animal

v/lth v/eight

1 4.3 gm. 6.5 gm.

2 0.7 " 7.1 "

3 3.5 " 7.7 "

4 4.8 " 8.6 "

5 3.0 " 7.1 "

6 4.8 " 6.7 "

7 4.8 " 7.9 "

8 3.8 " 8.5 "

9 2.0 " 3.5 "

10 4.8 " 3.8 "

11 3.0 " 5.0 "

I

34

Animals Ko. 2 and Ko. 9 showed the most rapid exhaustion with the least load. Some inherent weakness may have accounted for this as both animals died after three months. Animal l\o, 2 crept poorly from the beginning and it was possible' to obtain onl^;- half the number of runs as com- pared with the other animals when creeping v/ithout loads. The same v/as true for No. 9, except that great activity was shown for the series that was obtained, though only half the series as compared with the other animals was obtained. The results show that exhaustion caused by added weights was independent of the v/eight of the animal. The load which caused exhaustion for the majority of the animals v/as 3.0 gm. or more up to 4.8 gm.. Conseauently individual lead v/eights of 0.5 gm. v/ere chosen as the most suitable to obtain effects of small added loads up to 4.3 or 4.8 gm.

The method of applying the weights was as follov/s: the weight for each load v/as gradually increased but these increased loads were not added in immediate succession. A run without any added load or a load smaller than the load just used was made between the various increases. Table V illustrates this method. The advantage of this method was to insure the fact that any effects of fatigue caused by the additional weights could be avoided by having a run of no load or a lighter load than had just been removed before the next heavier weight had been added. This was the method used to obtain data for eleven

animals, the ranpie of temperature for the entire series being from 19,5° to 24.8°G. but constant for each e:x:peri .lent .

The way in which these graphs were made was to take the data from a group of animals, i.e., animals numbered 3, 4, 7 and 10 and use it to plot four different comparative graphs (each graph for each animal) showing the following relations :

a. Between velocity of progression and velocity of pedal wave.

b. Between freouency of waves and velocity of progression.

c. Between advance due to a single' wave and speed of creeping.

The measurements obtained vmen 8,300 waves were ob- served while eleven animals were carrying varying loads during vertical ascension were used in the analysis of the data. A comparison was made with and without load from the actual scatter of the data. The same relationships as those described for the last experiments were used as functions of one another. Collective graphs were plotted. The following typical graphs illustrate the way the data v/ere utilized throughout these experiments. Fig. 5a shows the actual scatter obtained from direct observations from a series of runs with and without load for animal l\o. 7. The relation between the velocity of progression and the frequency of waves has been plotted. Fig. 5 also shows the effect of added loads on animal Ko. 7 'vhen the freouency

I

56

TABLE V

Method of obtaining data v/iien varying loads v/ere added to one animal during vertical creeping.

Animal Series 10 Runs of Without Kdded 'jVith Added

10 Waves each Load Load

1 "

tt

2 "

0.7

gm.

(run

1)

w

4 "

1.2

tt

(run

1)

5 "

0.7

11

( run

2)

6 "

2.0

ji

7 "

ti

8 "

2.5

II

9 "

0.7

II

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5)

10 "

5.0

tt

11 "

\»2

ti

( run

2)

12 "

5.8

II

15 "

14 "

4.5

tj

etc. until exliaustion

37

of waves is plotted against the velocity of progression. However, this graph has been derived by first plotting the actual scatter for each added load (each load plotted on a separate graph). Then the average points of the actual scatter for each load were determined and the best fitted line drawn. A comparative graph v/as made of these lines - each line representing an added load. Fig. 53 also illustrates for animal No. 10 the relation between frequency of v/aves and the velocity of progression when creeping vertically upwards carrying varying loads.

From an analysis of the data obtained under the above conditions it is possible to study the individual effect of each load on the various relations. This indi- vidual effect can be observed by comparing the slope of each line for each load. For instance, in Fig. 5 these results show that this relationship (frequency plotted against velocity of progression) is independent of the load carried. This is strikingly illustrated when we analyze these slopes; for exam.ple Mo. 7, without load has a slope of 0.20, with 3.8 gm. added the slope became 0.22, and with 4.3 gm. it was again 0.20.

Pig. 6 illustrates the effect of tension as related to velocity of progression. No definite lav/s can be stated, but in general the first loads (0.7 gm. to 2.0 gm. ) tend to increase the velocity of progression. A load of 2.5 gm. tends to diminish the rate of creeping and v-ith increasing load usually there is a sli^uit increase in the

58

rate and then no change.

After the results were ueterti\lr.t.a as p;iven in Fip;. 6 a comparison was made to determine the probable error when plotted with var^z-ing loads. Factors v/hich account for error may be fluctuations in the velocity with v/hich the individual waves traverse the foot and the possibility that the adhesive pov.er of. the foot largely depends on its extruded mucous material v/hich may result in a slight error when comparing the effectiveness of the pedal mechan- ism.

The probable error of any one of the activities may be plotted against varying loads and then ' compared v/ith the normal. graph showing the relationship between the probable error for velocity of progression and varying loads is given in Fig. 7"^. The probable error appears to be greatest when weights of 0.7 gm.. , 1.2 gm. and 2.0 gm.. were added, and very slight with 3.0 gm. , 3.8 gm, and 4.3 gm. A large probable error in experiments of this kind would not be expected because the factors observed in these experiments are very definite, each wave being readily discernible.

Using Bess el's formula for probable error;

0.6745 , ~v

/\Jn (n-1)

the- follov/ing values were obtained and plotted against the varying loads. Animal Ko. 6.

V/ithout loads 3,04 With load 2,5 gm. 5.12

With load 0,7 gm. 4,34 , " " 3.0 " . 3.45

" " 1.2 " 3.96 " " 5.8 " 5.35

'» " 2.0 " 3.50 " " 4,5 " 5.45

39

Table VI shows the effect of loads on the individual factors involved in locomotion, i^bsolute values and' per- centage of increase or decrease during vertical ascensions with added loads are compared with ascensions v/ithout load. In analyzing the effects of these added loaas on the individual activities the freauency is found to snow the least alteration and velocity of progression the greatest.

The results of the foregoing experiments confirm for the first time for Helix these facts relative to loco;7iotion. Tiie results show that the velocity of a single wave, the frequency of the waves and the advance per wave as related to the rate of creeping are independent of the load carri3d. These facts show that the foot is an independent effector* This evidence indicates that the intrinsic neuro-muscular mechanism of the foot is the primary factor in locomotion. Cole, on the contrary, has stated (1925-26) that the stimulus for locomotion in Helix is tension of the body muscles produced by downward pull of gravity and that the stimulus is received by proprioceptors of these ::iuscles. This conclusion would indicate that the primarv fpctor in locomotion is the central nervous control v/hicu coes not accord with the experimental findings of the above experi- m.ents. Experiments reported later in this thesis show that the foot is secondarily under the control of central nervous" impulses but that primarily it is an independent effector.

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C. Vertical ascensions of the de-eyed Helix with and v/ithout loo.d.

A series of experiments was carried on to determine whether the lav/s of linear proportionality governing loco- motion would be altered if the animal were de-eyed, hs far as can be determined no mention has been made in the literature of experiments of this nature. A comparison was then made between the normal animals and the s&rn.e ani- mals de-eyed (Helix lactea ) . The eyes are easily renoved as they are situated at the tip of the posterior tentacles which are supplied by nerves from the supraoesophageal ganglia, i.n alteration in the velocity of the pedal waves, the velocity of .progression, the frequency of waves and the advance per wave, even though slight, might be expected in the de-eyed animal v/hen the posterior tentacles are cut off. De-eyeing should interfere with reflex interplay due to the connection of the nerve net found in Helix.

iiervous system in Helix. In the nervous system of mol- luscs there are some highly characteristic features. In Helix it consists of three pairs of ganglia associated with important sense organs and connected by nerve cords. One pair (Fig. A3,c) v/hich lies dorsal to the oesophagus supplies the tentacles and the eyes. Sometimes this cerebral ganglion is considered to be made up of two supraoesophageal ganglia joined by a broad transverse commisure. A second pair (Fig. A3,p) lies TFciitral - to the alimentary tract on the front part of the muscle mass of the foot. These are the pedal ganglia which are connected with the otocysts, consisting of two small sacs imbedded in the pedal ganglia. The third pair, the visceral ganglia (Fig. A3,v) are also ventral. The arrangement of the nerve-net can be seen from Fig. A2.

44

The supraoesophageal ganglia are connected with the sub- oesophageal ganglia, which consist of tv/o principal ganglionic masses. The forward mass is a pair of ganglia, the pedal ganglia, and the posterior mass consists of the visceral ganglia. Therefore, since the supraoesophageal and suboesophageal ganglia are connected it is expected that de-eying v/ould affect the reflex control of the pedal ganglia.

After measurements were obtained for the normal animals with and without loads v/hile creeping vertically upv/ard, the same animals were de-eyed. The temperature varied from 19.5*^ to 24.8°G. but v/as constant during each experi- ment."'" Data were gathered from the observation of 1100 waves for the normal and the de-eyed animals carryine- no load. For animals carrying various loads 2700 waves «ere obtained and measurements taken. Graphs showing the actual scatter obtained from these measurements and other graphs showing the average points of the actual scatter were plotted according to the m.ethod described for the previous experiments.

Table VII shows a comparison of the de-eyed and normal anim.al during vertical ascension with and without loads. Absolute values and the percentage increase or decrease are given for the velocity of a single pedal wave, the rate of creeping, the frequency of waves and the advance per wave. In a great many cases the individual activities

V.Tierever temperature limits are m.entioned it is to be understood unless otherv/ise stated that the temperature was constant for each experiment.

45

taking place during locomotion in the de-eyed animal are increased when compared with the normal. This increase may be caused by the removal of the eyes. Reference to Pigs. 8, 9 and 10 shows that the linear relation betv/een velocity of progression and frequency of pedal waves or that between speed and dimensions of a sing-le wave, or between the advance per wave and the rate of creeping is not altered in the de-eyed Helix lactea .

These facts are further illustrated by Figs* 8A, 9A and lOA. These shov/ the actual scatter obtained from direct observation of the normal and de-eyed animal. The relations between the velocity of a single pedal wave and the rate of creeping (Fig. BA), betv/een the velocity of progression and the frequency of waves (Pig. 9A) and between the advance per wave and the velocity of progression (Pig. lOA) have been plotted for animal No. 3 during vertical ascen- sions v^ithout load.

It was determined also froro a series of experiments that the carrying of added loads during vertical ascen- sions does not alter the linear proportionality between these relations in the normal and de-eyed animal. Figs. lOB, lOG and lOD show the actual scatter obtained from direct observations under these conditions (normal and de-eyed). The relation between the velocity of the pedal waves and the rate of creeping (Fig. lOB), between the velocity of progression and the frequency of waves (Fig. IOC) and between the advance per v/ave and the velocit^^ of

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..oeression (Fig. loD) has been plotted for animal No. 3 enuring vertical ascensions with added loads of 0.7 grama.

Fig. lOE and Table Vila illustrate that the linear relation between the velocity of a single pedal wave and the rate of creeping is not altered when the de-eyed animal (ITo. 4) is creepin.. upward carrying added loads.

TABLE Vila

Animal No. 4 Added load Series slope of line

0.7 gni. 1

0.7 " 2

0.7 " 3

1.2 " 1

0.26 0.24 0.24 0.52 0.24

1.2 " 2 ^•^ " 0.26

Q C ft

0.30

^•^ " 0.24 3«8 " 0.14

" 0.64

exhaustion

These data show that the foot is primarily an Independent effector. It may be secondarily under cen- tral nervous control as evidenced by the slight alteration

49

that occurs in the Individual activities wnen the eyes are removed.

The significance of the terra "independent effector" is shown by the fact that "for equal rates of creeping (Liiaax) the activity of the pedal organ is independent of the added loads. This is consistent .vith the fact that the activities Of the pedal organ are determined by its intrinsic neuro-muscular structure. The nervous elements (nerve-net) in the foot are secondarily under the control of the central ganglia." (^rozier and Federighi, 1924-85,0). Added loads do not alter the intrinsic activity of the creeping organ, but appear to act on the central reflex mechanism which inhibits or releases the pedal waves. Therefore we may speak of the foot so far as it concerns its production of pedal waves as an "independent effector" secondarily under the central nervous control, since the laws of its aotlvit- are the same regardless of the load carried in locomotion.

D. Investigation of the beliavior of the detached foot.

The foot v/as detached from the pedal ganglia to gain further information concerning the ad jus tor mechanism regulating locomotion. diagram showing (a) the nerve- net in the foot of Helix pomatia and (b) the principal parts of the nervous system is snown in Fig. 2.., taken from

Ubujigen aus der vergleichenden Pliysiologie", by Jordan (1927, p,22.5). By comparing the normal v/ith the pedal-free

50

anl^l Jordan h.s concluded that the principal lur.ctlon of the pedal gan^c^ia as v,ell as the sin^cle ganglion of the .-acidlana is the reg,.lation of "vlsoosoid" tonus, i-e con- cluded that the uniform condition of the musculature in the nornial animal is to be considered a sort of an eauill- brium between two processes, the peripheral and presu,^,bly reflex Production of resistance, and the steady lessening of this condition by the pedal ganglia.

The following experiments on twenty-four animals were performed to study tne oenavior of the foot when separated from the pedal ganglia. A series of anim.ls was used in Which the foot n.d neen cut directly fror. .ne visceral organs. Immediately after the snail had been creeping actively the foot was held aw,y from th. shell by means of a forceps placed as near t.., visceral organs as possible, and then the foot was cut off quickly.

The isolated foot -.vas cttacied at one end by silk thread to c. ..-ar;; lever ana at the other end to an L-snaped rod (Fig. 11) which was held in place by a muscle-clamp. A moist chamber surrounded the foot. J.ymograph records were ootained snowing the action of the foot. Tne limits of temperature for the entire series of experiments were from IS. 5° to 24.8°C;.

Reference to Figures 11a &nd lib shows that the usual rhythmical action of the foot is rre^.tl-r interfere- when it is not conneote. .vlsn tae pedal ganglia. Even in kyinograms showing some recurring waves the usual periodic

51

recurrence found In the intact animal is destroyed. The destruction of the periodic recurrence of .avea when tne pedal ganglia are separated from the foot suggests that this periodicity is determined by the presence of these ganglia.

Extirpation of the pedal ganglia in the intact anirnal may be possible and further study of the locomotor waves under these conditions may bring to light additional infor- mation as to the exact action of the pedal ganglia in this neuro-rnuscular activity.

E. The effect of mechanical stimulation and of Ringer's solution on locomotion.

Preparatory to experiments Involving the injection of certain drugs a series of control experiraents ..vas performed on Helix lactea and Helix pomatia. The controls ,vere tested for (a) consistency of runs, (b) the effect of mechanical stimulation, (o) the effect of Klnger's solution (cold blooded) .

In testing the consistency of the runs a comparison was made of the data obtained for runs ^ separated by intervals of time. Reference to Table VIII sho^.vs that in a ..riven ex- periment the data obtained at the end of three hours may vary slightly from the data obtained at the end of one or

^ The composition of Ringer's solution (1000 cc. of --ter)

used m every case was as follows- .^ter)

Grams

i:aCl 6.5

KCl 0.14

CaClg 0.12

I^'aHC03 0.20

52

at the end of two nours. Except fop this slir^^t variation the results ms.y be said to be consistent.

The controls ivere also tested to determine the mechan- ical stimulation on vertical creeping. mien tlie animal was creeping actively upwards a hypoderr.iic needle, v/hich was to act as the mechanical stimulus, was inserted into either the body v/all in the region where the visceral mass joins the foot or into the anterior end of the foot ^.ir-ctlv back of the tentacles. The reactions of the animal were noted.

The method followed in these experiments is illus- trated by the follov/ing, v.-hich represent the procedure of a typical experiment in this series v/hen the animal was creeping upward.

(a) Records of norm.al run obtained.

(b) Records taken of runs after mechanical stimulation of the body wall.

(c) Interval allowed to remove the effects of this stimulation.

(d) Further records of normal run.

(e) Records of runs obtained after mechanical stimu- lation of anterior end of the foot.

In eight out of ten animals used the mechanical sti:au- lation of the body wall caused a withdrawal of the head into the shell, followed by a slight contractio:-^ -^f -^':e foot; then creeping began immediately. In one of the other two cases (experiment 50, animal No. 15) creeping did not

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55

begin for 43 minutes after stimulation. In the other instance (experiment 52, animal IIo. 16) it did not begin for nine minutes after pricking.

Llechanical stimulation of the body wall also caused an increase in the velocity of progression and in the ad- vance per wave in Helix pomatia (Table Villa), ar^ either a decrease or an increase in the frequency of waves and in the velocity of a single wave. No definite conclusion can be drawn, however, from this type of experiment on Ilelix lactea (Table Vlllb), for in some of the experiments these activities were increased, while in other experiments they were decreased.

Table VIIIc shows the effect of mechanical stimulation at the anterior end of the foot does not differ greatly from, stimulation of the body wall. Here again the results do not show any definite consistency, the frequency of waves and the velocity of a single wave being least affecte Therefore the mechanical effect can be eliminated in the subsequent experiments.

The effect of Ringer's solution was tested on the controls. Experiments were carried out by injecting 0.2 cc. of Ringer's solution and studying the effects. Data for these experiments are given in Table Vllle.

As the chemical composition of the blood of inverte- brates approaches that which we find in the lower verte- brates, amphibian (or cold-blooded) Ringer's sol.;;.ijn c_us the drug was used in the following experiments.

56

TABLE Villa

The effect of mechanical stimulation of the body wall of Helix pomatia. Absolute values are given for the fac- tors involved in locomotion.

Ko. of Velocity of

i-'requency

Velocity of

iidvance

Experiment progression

oi waves

single v/ave

per wave

in mm. per

per m.inute

in mm. per

in mm.

minute

iriinut e

5 (Normal)

8.5

42.0

3353

0.24

6 (Mechanical

stimulation)

14.8

43.0

3525

0.33

10 (Normal)

19.2

71.4

6900

0.26

11 (I.iechanical

stimulation )

23.0

62.0

6630

0.56

14 (Norm.al)

13.4

59.5

82i00

0.22

15 (Mechanical

stimulation )

19.0

52.0

7300

0.36

20 (Normal)

12.2

62.5

7600

0.19

21 (Mechanical

stimulation)

18.6

83.3

6700

0.22

57

TABLE VII lb

of

The effect Helix lactea

of mechanical stimulation on the , on vertical creeping.

body wall

No. of Velocity of experiment progression in mm. per

minute

Frequency of waves per minute

Velocity of sin^f^le wave in mm. per

minute

i-.dvance per t»ave in mra.

41 42

(Normal ) (Lie Chan ical stimulation )

22,0 27.7

62.5 62.5

4500

4300

0.35

0.44

48

\ IV X ilicL J- 1

(i-iechanical stimulation)

24.0

62.5 83. 0

4900 4700

0.24 0.29

49 50

(Normal )

(Mechanical

stimulation)

23.0 15.0

71.0 83.0

5700 5100

0.32 0. 19

51 52

(Normal) (Mechanical stimulation )

48.0 40.0

83.0 71.0

4900 5100

0.57

72 75

(Normal)

(Mechanical

stimulation)

20.5 15.3

83.0 83.0

4400 4000

0.24 0.18

78 79

(fiormal )

(Mechanical

18.6

r;3.0

4600

0.22

stimulation) 19.6 83.0 5000 0.23

58

TABLE VI lie

The effect of nechanical stimulation of the anterior

end

of the foot

of helix

lactea on

vertical creeping.

No.

of Velocity of

Frequency

Velocity of

Kdvance

experiment progression

of waves

single wave

per wave

in mm. per

per minute

in mm. per

in mm.

minute

minute

43

(Normal )

29.0

100.0

4100

0 . 29

A A

44

(Mechanical

stimulation)

17.0

(DO . \J

40UU

U . cx

47

(Normal )

18.3

83.0

4/00

0 . 22

4c5

(i.Iechanical

stimulation )

l«i) . 8

oo . u

ATy r\r\ 4 / UU

0 . lb

53

(Normal )

14.8

71.0

4yuu

U • (dU

54

(Mechanical

stimulation )

22.0

DO . u

U • /i o

55

(Normal )

38.0

125.0

6025

0.30

56

(I.Iechanical

stimulation )

13.6

72.0

4200

0.18

74

(Normal )

15.5

131.0

5550

0.18

75

(Mechanical

stim.ulation )

13.6

72.0

4200

0.18

80

(Normal )

44.0

122.0

6000

0.34

81

(laechanical

stimulation )

20.9

125.0

6375

0.17

TABLE VI I Id

Effect of doses of Rinrer's solution (0.2 cc. on the velocity of progression (Y), frequency of waves (F), velocity of pedal waves (v) and the advance per wave (A).

Kelix lac tea Plus"^ Minus Cases unchanged

V 9 10 1 P 8 ■ 12

V 8 12

A 6 10 4

Increase or decrease from the normal animal after injection of Ringer's solution, giving total number of cases of increase or decrease respectively

60

TABLE Vllle

The effect of Ringer's solution on vertical creeping.

Ko. of

Animal

Velocity of progression in mm, per minute

Frequency of waves per minute

Velocity of single wave in rnm. per minute

r.dvance per wave in mm.

1

Norma 1 Ringer ' s

38.8 47.4

38.8 37.3

4800 4700

0.36 0.49

2

Normal Ringer ' s

33,1 38.9

85.4 70.3

5040 5100

0.47 0.52

3

Normal Ringer ' s

30.9 24.3

75.1 68.6

4450 4370

0.41 0.29

4

Normal Ringer ' s

20.4 13,1

69.8 65.2

4900 5300

1.09 1.0

5

Normal Ringer ' s

43.1 25,1

99.4 79.1

5680 5920

0.38 0.31

6

Normal Ringer ' s

21.1 23.2

68.8 63.8

5550 4440

0.31 0.24

7

Norm.al Ringer ' s

35.9 22.6

83.5 81.6

5950 5720

0.39 0.52

8

Normal Ringer ' s

21.0 15.4

70.6 72.1

4200 3880

C.32 0.54

9

Normal Ringer * s

36.6 39.8

80.1 106.6

5220 4940

0.46 0.37

10 Normal Ringer ' s

29.4 43.7

58.3 71.1

3980 4720

0.16 0.35

61

TABLE VI He

(cont, )

"Ko . of Animal

progression in mm. per

m.inute

rx'ecu.ency of waves per minute

veiociLy 01 single wave in mm. per

minute

i-.dvance per wave in mm.

11 Normal Ringer ' s

9.7

33.7

82.1 57.6

6560 4020

0.39 0.22

12 Normal Ringer ' s

18.1 29,2

75.0 84.2

4200 4420

0.27 0.35

13 Normal Ringer ' s

53.8 33.1

94.1 75,9

5140 4520

0.57 0.44

14

Normal Ringer ' s

27.3 34.1

90,2 91.6

5640 4360

0.32 0,37

lb Normal Ringer ' s

28.1 19.3

65,0 66,4

4800 4580

0.28 0.19

± D

Normal Ringer ' s

19.1 16.1

77,0 61.0

4000 5060

0.21 0.20

JL f

Normal Ringer ' s

12.7 12,8

69.4 73.3

4800 4450

0.18 0.18

lo Normal Ringer ' s

41.1 16.4

82.8 71.5

4560 4900

0.23 0.24

19 Normal Ringer ' s

29.9 31,1

123.9 97.9

4980 3470

0.48 0.32

20 Normal Ringer ' s

21.4 18.6

84.7 91.7

4300 4420

0.20 0.21

62

F, The effect of adrenalin on the neui-o-.-nuscuiar activitie during locomotion.

The action of adrenalin on various kinds of tissue, particularly on the hearts of vertebrates, has been ex- tensively studied. Clark (1927) has cited references ■ (p. 62) concerning the effects of adrenalin on some inver- tebrate tissues, i.e., the intestine of the crayfish, the oesophagus of i^phrodite, of i-.physia and of Helix. He has also drawn attention to the fact that the inhibitory action of adrenalin is as common as its augmentor action in vertebrates, but that only augmenter action has been described in invertebrates. Boyer (1926) found that adrenalin and spartein slackened the rhythm of the isolated heart of the snails. He found that adrenalin affects neither the amplitude nor the tonus in the doses used (1 part in 10,000 and 1 part in 1,000), whereas spartein reinforces the amplitude and augments the tonus. V/yman and Lutz (1930) have described an inhibitory •:.c':ion of adrenalin on the isolated holotnurian cloaca. Their experiments demonstrated that adrenalin has both inhibitory and excitatory effects on invertebrate tissues.

Ho reference in the literature as far as can be determined has been made to any work on the effects of adrenalin on the muscles involved in locomotion of molluscs The following experiments describe the action of different amounts of adrenalin in the intact animal (Helix). Con- trasted with the study of the detacuea foot, whic:^ i./.o^vea.

63

interference with the circulation, the subsequent experi- ments show the effect on locomotion, i^drenalin, which has a low molecular weight, is easily diffusible, is not attached to protein and is readily poured into the blood stream from which it is removed by the tissues. Usually there is no delay in the manifestation of its effect. Its maximal duration of effect in mam-mals is generally only a few minutes and its physiological activity during this interval is narked (Kendall, 1929). However, in cold- blooded animals the effect of adrenalin m.ay be prolonged. Bieter and Scott (1929) reported a rise of blood pressure in Rana pipiens following a dose of 0.2 cc. of epinephrine hydrochloride, 1:10,000, which persisted for at least one and one-ouarter hours after injection. MacKay (1931) ■':5.s found that adrenalin causes rise of ventral aortic jlood pressure in skates (Kaia) , which persists from one to two and one-half hours. vVyman and Lutz (1932) reported that intravenous injection of adrenalin in Squalus acant.-ias produced a rise of both the ventral and the dorsal aortic blood pressure, both systolic and diastolic, persisting for at least thirty minutes. .-.Ithough not a great deal of work has been done to compare the lasting effect of adren- alin in the cold-blooded animals v.'ith the effect in m.aminals the evidence so far obtained shows that there is more pro- longed maximum effect in cold-blooded animals.

Adrenalin chloride solution (Parke, Davis viV Co.) .vas used in some of these experiments. In others dry suprarena

64

extract (Parke, Davis a Co.) usea Ir^ oi'aer to check

the results obtained from adrenalin c hloride because of HGl and chlorotone.

The same general procedure was followed as previously- described. An investigation was made to see whether the linear proportionality between the various relationships for vertical creeping held when adrenalin was injected. Data were obtained from 68 animals. The results found for Plelix lactea confirmed those obtained for Helix pomatia. Therefore, since Helix lactea exhibits more active creeping and lives in the laboratory for longer periods of time than Helix pomatia , Helix lactea was chosen for the majority of experiments.

Data were first obtained for vertical ascensions

under norm.al conditions. Then these data v/ere compared

with the data obtained after adrenalin (of a particular

concentration) had been injected. The data p-iven below

were obtained from a series of eighty experiments. From

the 68 animals used a total of 40,500 waves were observed,

i.e. , 19,700 for the normal animal and 20,800 for the

injected animals. Adrenalin chloride solution (Parke,

Davis & Co.) was added to Ringer's solution to make

various concentrations, i.e., 1:10,000, 1:20,000, 1:40,000, 1:100,000 ,

1:80,000/1:120,000 and 1:200,000. Measurements were first obtained for vertical creeping under normal conditions. Then 0.3 cc. of adrenalin (of the concentration to be studied) was injected into Helix pomatia , and when lielix

65

lactea was used 0.22 cc. of adrenalin was injected. The time elapsing between the injection and the resumption of creeping varied in different animals from immediate creep- ing to one hour. In a few cases creeping did not begin for two to three hours.

In order to determine whether the linear propor- tionality between the various relationships for vertical creeping held when adrenalin was injected the measurements obtained for the normal and injected animal were employed as described in the previous experiments. Graphs v/ere plotted of the average points showing the relation betv/een the velocity of a single pedal wave and the velocity of progression, between the frequency of waves and tlie velocity of progression, and between the advance per wave and the velocity of progression. The best fitted line was dra';m and the slope of the line determined. The slope of the line obtained for a particular relation for the control was com.pared with the sam.e relation for the injected animal. Figures 12, 13 and 14 show the typical results. It is noted that the straight line proportionality and the slope of the line are essentially unchanged by adrenalin. These are added features which support the idea of an "independent effector in muscle.

A description is given below of the effect of injection of various concentrations of adrenalin chloride. Table IX shows the effect of 0.2 cc. of adrenalin (1:10,000) on Kelix lactea during vertical ascensions. i:drenalin of

56

this concentration usually causes a decrease in f^e velocity of progression, in the frequency of waves, in the vcj-ocI:/ of a single wave and in the advance per wave.

Similar experiments were performed on Helix pomatia (Table IXa ) . Prom 1950 waves observed on the normal animal and 2000 on the injected animal data were procured from, which Table IXa has been computed. Adrenalir. (0.3 cc. of adrenalin, 1:10,000) had the same effect on these animals as on Helix lac tea , i.e. , a decrease in each of the factors involved in locomotion, during vertical ascensions. Table X shov/s that the same results in the main are obtained with both species.

Tables Xa through Xh give the data from which Table X was compiled. Other experiments employing adrenalin in doses of 1:20,000, 1:40,000, and 1:80,000 (Table XI) give the same results as obtained for doses of 1:10,000, i.e., usually & decrease in the velocity of progression, the frequency of waves, the velocity of a single wave and the advance per wave. With doses of adrenalin 1:100,000 (Table XIa) inconsistent results were obtained, sonetimes an increase in the activity and som.etimes a decrease in the same activity.

However, doses of adrenalin 1:120,000 (Table XI) produced a stimulating- effect on the velocity of progression, the frequency of v/aves, ti.e velocity of a si;.;jj.c! .vave •j.U'J. sometimes either an increase or decrease in the advance per wave.

67

TABLE IX

Effsct of injection of adrenalin chloride (0.2 cc. 1:10,000) on the velocity of a single wave (v), the rate of creeping (V), the frequency of waves (F), and the advance per wave (A) (Helix lactea ) during vertical ascensions. *

Ko. of

animal

linn « rum.

V =

min.

p _ waves mm.

— mm.

waves

2

0 = control • = injected

2576 2682

36,6 34.0

66,0 67.8

0.33 0.29

3

0 = control 0 = injected

3186 3060

45.2 44.7

76.0 67.2

0.38 0.34

7

0 = control • = injected

5381 4200

30.4 28.1

83.4 80.1

0.34 0.36

9

0 = control 0 = injected

.3334 4704

99.6 55.2

79.2 77.4

0.46 0.36

10

0 = control • = injected

4660 4358

30.3 23.7

64.6 88.2

0.46 0.29

13

0 = control • = injected

6080 5250

24.4 25.1

74.3 60.7

0.33 0.42

14

o = control • = injected

3908 3510

55.3 43.2

79.2 72.0

0.40 0.34

68

TABLE IXa

A comparison of the normal and injected anim.als''' (Helix poma tia ) during vertical ascensions. Absolute values for the velocity of a single wave, the rate of creeping, the frequency of waves and the advance per wave.

Ko. of

animal

Ve a

in

locity of single v/ave

mm. per minute

Rate of creeping mm. per minute

Frequency in waves per

of mm.

iidvance per wave in mm.

60 Control Injected

6920 5400

48.2 17.2

96.0 92.0

0.96 0.24

62 Control Injected

7830 5640

46.8 22.5

103.1 81.3

0.45 0.28

66

Control In j ected

6010 4950

19.2 36.0

95.2 92.0

0.19 0.38

69 Control Injected

5770

8800

45.1

29.9

60.0 87.0

0.78 0.50

70

Control Injected

6280 6460

21.9 19.6

62.2 94.0

0.34 0.31

75

Control Injected

6350 5210

33.5 18.9

71.9 62.0

0.45 0.32

Injection

of 0.3 cc.

adrenalin

chloride, 1:10,000

•

69.

TABLE X

Effect of doses of adrenalin (1:10,000) on the velocity of progression (V), frequency of waves (F),

the

velocity

of pedal waves

(v), and the

advance

wave

(A).

Helix

lactea

Helix

pomatia

Plus^

Minus

Plus

Minus

V

1

6

v'

1

5

P

2

5

F

2

4

V

2

5

V

2

4

A

2

5

A

1

5

Increase or decrease from, the normal animal after injection of adrenalin (1:10,000), giving total number of cases of increase or decrease respectively.

70.

TABLE Xa

Effect of doses of adrenalin chloride (1:10,000) on the velocity of progression (flelix lactsa).

l\o. of

Velocity of

Velocity of progression

in mm. per minute

Control

After dose of adrenalin

1: 10,000

2

36.6

34.0

3

45.2

44.7

7

30.4

28.1

9

99.6

55.2

10

30.3

23.7

13

24,4

25.7

14

55.3

■ 43.2

TABLE Xb

Effect

of doses of adrenalin chloride (1:10,000)

on the velocity of progression (Helix pomatia).

llo. of

Velocity of

Velocity of progression

experiment

progression

in mm. per minute

in mm. per minute

after dose of adrenalin

Control

1: 10,000

60

42.2

17.2

62

46.8

25.5

66

19.2

36.0

69

45.1

25.9

70

21.9

19.6

75

33.5

18.9

i

1

71.

TABLE Xc

Effect of doses of adrenalin chloride (1:10,000) on the frequency- of waves (Helix lactea ) .

No. of

A y "n p "P "1 TTi PTi "h

Frequency of waves •r)c.-p minntp'

Control

Frequency of waves

•np*p Trni'Tii'i'hp

III JUXILA.

After dose of adrenalin 1:10,000

2

66.0

67.8

3

76.0

67.2

7

85.4

80.1

9

79.2

77.4

10

64.6

88.2

13

74.3

60.7

14

79.2

. 72.0

TABLE Xd

Effect of doses of adrenalin chloride (1:10,000) on the frequency of waves (Helix pomatia )

Ko. of Frequency of waves Frequency of waves

experiment per minute per minute

Control After dose of

adrenalin 1:10,000

60 96.0 92.0

62 103.1 81.3

66 95.2 92.6

69 60.0 87.0

70 62.2 94.0 75 71.9 62.0

72.

TABLE Xe

Effect of. doses of adrenalin chloride (1:10,000) on the velocity of a single v/ave (Helix lac tea ) .

No. of

exper iaient

Velocity of a sinp:le wave in rmn. per minute

Control

Velocity of a single wave in mm. per

minute >-.fter dose of adrenalin 1:10,000

2

2576

2682

3

3186

3060

7

5381

4200

9

3334

4704

10

4660

4358

13

6080

• 5250

14

3908

3510

TABLE Xf

Effect of doses of adrenalin chloride (1:10,000) on the velocity of a single wave (Helix pomatia ) .

i:o. of

experiment

Velocity of a single wave in mm. per minute

Control

Velocity of a single wave in mm. per

m.inute After dose of adrenalin 1:10,000

60 62 66

69 70 95

6920 7830 6010

5770 6280 6350

5400 5640 4950

8800 6460 5214

73

TABLE Xg

Effect of doses of adrenalin cnloride (1:10,000) on the advance per wave (Helix lactea).

No. of

experiment

Advance per wave in mm.

Control

advance per wave in mm.

After dose of adrenalin 1:10,000

2

0.33

0.29

0.38

0.34

7

0.34

0.36

9

0.46

0.36

10

0.46

0.29

13

0.33

0.42

14

0.40

0.34

TABLE Xh

Effect of doses of adrenalin chloride (1:10,000) on the advance per v/ave (Helix pomatia ) .

No. of

experiment

Advance per wave in nim.

Control

60 52 66 69 70 75

0.96 0.45

0.19 0.78 0.34 0.45

Advance per wave in !nm.

After dose of adrenalin 1: 10,000

0.24 0.28

0.38 0.50 0.31 0.32

74.

TABLE XI

Summary of effect of various concentrations of sdrenalin

on factors involved in locomotion.

Velocity of

Dose

1 of

adrenalin

!Number of cases

of increase

progression

or decrease from

normal

Plus

iviinus

0. 2

c c .

1: 10,000

1

6

0.3

cc .

1: 10,000

1

5

0.2

c c .

1 : 20 . 000

1

6

0.2

cc .

1:40, 000

2

4

0.2

cc .

a.: 80,000

3

13

0.2

cc .

1: 120,000

8

1

r I tJ u Lit? lio y ux

L> •

p

o

W ci V O O

O U •

p

0.2

cc.

1: 20,000

2

5

0.2

cc .

1: 40,000

1

5

0.2

cc .

1:80,000

2

14

0.2

cc .

1: 120,000

6

3

Vp 1 or i fv o f*

0.2

c c •

1: 10 .000

4

9

i3 J- t J. ti, JL w <V CL V ^

^ .

1*10 000

p

4

0.2

CC.

1:20,000

2

5

0.2

cc .

1:40,000

1

5

0.2

cc .

1 : oO , 000

14

0.2

cc .

1: 120,000

6

3

advance per

0.2

cc .

1: 10,000

2

5

wav e

0.3

cc .

1: 10,000

1

5

0.2

cc .

1:20,000

2

5

0.2

cc .

1: 40,000

1

5

0.2

cc .

1: 80,000

4

11

0.2

cc .

1: 120,000

5

4

75.

Recapitulation of TABLE XI

1: 10

,000

1:20

,000

1: 40

,000

1:80,000

1:120,000

+

+

+

+

+

Velocity of progression

2

11

1

6

2

4

8 1

Frequency of waves

4

9

2

5

1

5

7 9

8 1

Velocity of a single wave

4

9

2

5

1

5

2 14

^ IV

Advance per wave

3

10

2

5

1

5

4 11

5 4

TABLE XIa

Effect of doses of adrenalin chloride (1:100,000) on the velocity of proscression, frecuency of v/aves, velocity of a single wave and advance per wave.

In U . O I

vexoGiuy QI

Velocity of progression

experiment

progression

in mm. per minute

in mm. per minute

After dose of adrenalin

Control

I : 100, 000

o c,

oo • c,

45. 1

3

32.3

69. 1

18

68.0

80.8

21

42.0

40.4

28

81.3

47.1

Frequency of waves

Frequency of waves

per minute

per minute

After dose of adrenalin

Control

1: 100,000

2

95.2

90.1

3

78.1

121.0

18

64.5

95.5

21

71.1

109.3

28

99.0

83.0

Velocity of a single Velocity of a single ?/ave in min.per minute ' .^'ave in mm. per minute

after dose of adrenalin

Control

1; 100,000

2

4500

5120

3

5150

5500

18

5340

4420

21

3600

4500

28

6220

5930

Advance per wave

advance per wave

in mm.

in nun.

j.fter dose of adrenalin

Control

1: 100,000

2

0.40

0.46

3

0.41

0.48

18

1.18

0.65

21

0.63

0.73

28

0.61

0.47

ill

77

In order to determine whether the above results v/ere due to the action of adrenalin and not to the chlorotone present in the adrenalin chloride solution or a change in Hydrogen ion concentration, dry suprarenal extract (Parke, Davis & Co.) was nade up with Ringer's solution, A series of experiments was carried out under the same conditions as when adrenalin chloride was used. TablesXIIa through Xllh give the data obtained when adrenalin chloride 1:80,000 and suprarenal extract 1:80,000 were used respec- tively, and Tables Xllla through Xlllh give the data for experiments when adrenalin chloride 1:120,000 and supra- renal extract 1:120,000 v/ere used. A summary of this comparison is given in Table XIV. Reference to this table shows that results obtained from, suprarenal extract were the same as for adrenalin chloride solution, except that a greater depressant action was observed on the frequency of waves with suprarenal extract of doses of 1:80,000. The action of doses of 1:120,000 adrenalin chloride and 1:120,000 suprarenal extract was to cause either an increase or decrease of the advance per wave.

Injections of adrenalin of any concentration within the limits used, i.e., 1:10,000 to 1:120,000 during ver- tical creepinp: did not usually affect the number of v/aves present at one time on the foot. If there was a change in the number it was by having one more wave present after the injection of adrenalin. It was also consistently ob- served after injection of adrenalin of either strong or

78

TABLE Xlla

Effect of doses of adrenalin chloride (1:80,000) on the velocity of progression.

Ko. of velocity of progression Velocity of progression

experiment in ram. per minute in ram. per minute

i-fter dose of adrenalin

Control

1:80,000

8

23.8

14.0

10

64.5

35.0

11

35.0

16.0

,12

34.2

20.6

13

16.1

15.9

14

17.2

16.0

15

41.1

36.6

16

25.6

33.8

17

34.9

30.8

18

25.6

33.8

19

39.6

29.7

20

34.9

30.8

21

41.0

37.2

22

49.2

36.7

23

46.1

28.1

24

25.3

33.4

25

53.6

58.0

79.

TABLE XI lb

Effect of doses of adrenalin chloride (1:80,000) on the freauency of v/aves.

^o, of Frequency of waves Frequency of waves

experiment per minute per minute

i-^fter dose of Control adrenalin 1 ? P,f

8

75.5

• 71.4

10

135.0

87.6

11

87.0

83.0

12

62.8

63.0

13

62.5

61.5

14

69.6

60.1

15

82.1

61.0

16

59.0

72.0

17

70.8

106.6

18

59.0

72.0

19

74.3

65.5

21

72.5

60.0

22

104.0

72.4

23

75.4

78.0

24

61.5

70.0

25

90.7

94.1

80.

TABLE XIIc

Effect of doses of adrenalin chloride (1:80,000) on the velocity of a single wave.

of

Velocity of a

Velocity of a

iriment

single wave in

single wave in

mm. per minute

mm. per minute After dose of

Control

adrenalin 1:30,000

8

4600

3900

10

5160

4416

11

4800

4700

12

6300

5430

13

5620

5340

14

4450

3300

15

6230

5980

16

6250

5680

17

5060

4740

18

6250

5680

19

5700

4580

21

6600

7000

22

5340

4820

23

6080

5900

24

4150

4120

25

5580

5680

81.

TABLE Xlld

Effect of doses of adrenalin chloride (1:80,000) on the advance per v/ave.

No. of

experiment

Advance per wave in ram.

Control

Advance per wave in mm. After dose of adrenalin 1:80,000

8

0.31

■ 0.17

10

0.48

0.43

11

0.42

0.20

12

0.55

0.23

13

0.24

0.24

14

0.24

0.29

15

0.44

0.51

16

0.44

0.46

17

0.49

0.29

18

0.44

0.46

19

0.52

0.36

21

0.58

0.41

22

0.47

0.45

23

0.62

0.41

24

0.41

0.48

25

0.70

0.60

82.

TABLE Xlle

Effect of doses of suprarenal extract (1:80,000) on the velocity of progression.

"No o"^ experiment

progression in mm. per minute

Control

Vp"! op "i "h V nf*

progression in

mm., per minute

After dose of suprarenal

extract -1:80,000

124

26.4

28.5

136'

39.4

43.0

138

26.7

18.2

139

30.7

17.3

145

38.5

19.0

146

52.0

13.1

147

36.1

12.8

150

23.9

26.6

151

41.1

13.6

155

45.9

33.0

156

17.7

14.8

83.

TABLE Xllf

Effect of doses of suprarenal extract (1:80,000) on the frequency of waves.

^o. of Frequency of waves Frequency of waves

experiment per minute per minute

Control

renal extract 1

124

79.1

78.0

136

76.6

61.2

138

82.3

79.1

139

52.8

66.1

145

64.6

58.6

146

92.6

90. 5

147

74.2

66.5

150

59.7

63.5

151

68.7

69.8

155

106.0

76.6

156

67.7

53.3

84.

TABLE XI Ig

Effect of doses of suprarenal extract (1:80,000) on the velocity of a single wave.

No. of

Velocitv nf n

vexoGiLy 01 a

experiment

single wave in

single wave in

ram. per minute

mm. per minute

Control

After dose of suprarenal

extract r:80,000'

124

4010

4260

136

5100

5200

138

4360

4380

139

5500

4400

145

6040

5700

146

6216

5400

147

4600

3600

150

4400

3930

151

4900

3200

155

4520

4460

156

4550

3510

85.

TABLE Xllh

Effect of doses of suprarenal extract (1:80,000) on the advance oer v/ave.

No. of Advance per wave .^-dvanct; per v;ave

experiment in mm, in mm.

i-.fter dose of supra- Control renal extract 1;80,000

124

0.34

- 0.39

136

0.60

0.68

138

0.33

0.18

139

0.49

0.22

145

0.60

0.36

146

0.75

0.14

147

0.48

0.16

150

0.40

0.25

151

0.68

0.20

155

0. 56

0.36

156

0.26

0.29

86.

TABLE Xllla

Effect of doses of adrenalin chloride (1:120,000) on the velocity: of progression.

Ko, of experiment

Velocity of progression in mm. per minute

Control

Velocity of progression in mm. per minute After dose of adrenalin 1:120,000

7 ,

14.6

19.6

10

10.7

37.1

11

33.0

50.5

12

43.0

37.0

13

37.0

42.6

14

26.1

38. 1

21

14.3

19.0

25

10.2

12.8

26

26.0

38.1

87

TABLE XII lb

Effect of doses of adrenalin chloride (1:120,000) on the frequency of waves.

^o* Frequency of v/aves Frequency of waves

experiment per minute per ?-ainute

After dose of

Control adrenalin 1:120^000

7 59.4 80.0

10 71.1 84.8

11 VI. 3 97.3 12- 68.1 127.0

13 126.6 91.2

14 70.0 100.0 21 58.6 84.7

25 52.3 62.0

26 70.6 100.1

88.

TABLE XIIIc

Effect of doses of adrenalin chloride (1:120,000) on the velocity of a single wave.

of Velocity of a Velocity of a

experiment single wave in single wave in

mm. per minute mra.^'per minute

After dose of adrenalin

Control

■ 1:120,

7

5370

5730

10

4100

4600

11

6400

7030

12

6300

7990

13

7250

5850

14

5300

4700

21

3704

4880

25

3220

3760

26

5300

4720

89.

TABLE XI I Id

Effect of doses of adrenalin chloride (1:120,000) on the advance per wave.

1:0. of

r-dvance per

wave .-.dvance per "vave

experiment

in mm.

in mm.

After dose of adrenalin

Control

1: 120,000

7

0,25

0.23

10

0.16

0.43

11

0.52

0.50

12

0.58

0.30

13

0.51

0.45

14

0.23

0.29

21

0.23

0.19

25

0. 18

0.16

26

0.23

0.29

90.

TABLE XI He

Effect of doses of suprarenal extract (1:120,000) on the velocity of pror3:ression.

No. of

experiment

Velocity of progression in mm. per minute

Control

Velocity of progression in mm. per minute After do3e of suprarenal extract 1:120,000

36

47.1

49.2

38

37.2

52.6

40

23.6

54.2

42

23.4

48.6

44

25.2

48.2

45

50.1

65.1

91.

TABLE Xlllf

Effect of doses of suprarenal extract (1:120,000) on the freouency of waves.

Ko. of Frequency of Frequency of waves

experiment waves per minute per minute

After dose of supra- Control renal extract 1:120,000

36 100.0 102.0

38 71.2 110.7

40 65.5 89.8

42 79.7 88.4

44 63.4 93.0

45 82.3 112.1

92.

TABLE Xlllg

Lffect of

doses of suprarenal

extract (1:120,000)

on the velocity

of a single wave.

Ko. of

experiment

Velocity of a Velocity of a single wave in single wave in mm. per minute mm. per minute

After dose of supra- Control renal extract 1:120,000

56

5604

6055

58

4540

5700

40

5840

5420

42

4660

4900

44

57 50

4890

45

4250

5020

93.

TABLE Xlllh

Effect of doses of suprarenal extract (1:120,000) on the advance per wave.

Ko. of

Advance per

v/ave Advance per 'Nave

experiment

in ram.

in mm.

After dose of supra-

Control

renal extract 1:120,000

36

0.47

0.48

38

0.52

0.47

40

0.34

0.26

42

0.54

0.27

44

0.29

0.39

45

0.33

0.38

94.

TABLE XIV

Effect of doses 1:80,000 Effect of doses 1:120,000

Adrenalin Suprarenal adrenalin Suprarenal

chloride extract chloride extract

Plus Minus Plus i.Iinus Plus i.Iinus Plus l.Iinus

Velocity of

progression 3 13 38 81 60

Frequency of waves 7

8

8

0

Velocity of a single wave

14

8

0

Advance per v/ave

4 11

3 8

5 4 3 3

95.

weak concentration during vertical creeping that the ten- dency to orient had been greatly lessened. previously mentioned orientation and locomotion in Helix are closely allied (Grozier and Navez, 1930) and further work is neces- sary to disentangle the neuromuscular control of these mechanisms •

Since there is no sympathetic nervous system in Helix the usual explanation concerning the point of action of adrenalin cannot be given. Adrenalin affects different types of muscle differently. In structure the muscles of the foot closely resemble other invertebrate muscle (Mendel and Bradley). Elliott (1905) stated that adrenalin does not excite the muscle fiber directly but acts on a substanc at the m.yoneural junction. He stated that all other sub- stances but adrenalin evoke, from, the group of plain m.uscle tissues when stimulatin'i them directly, a reaction differin only in degree and not in kind in each tissue. It is the peculiarity of adrenalin to cause sharp contraction in one and relaxation in the other. The cause is the same, the effects different. Therefore the reacting substance must be different and he has decided that since mechanical and chemical stimuli do not point to marked intrinsic differ- ences in the plain muscle fibers they do reveal differences in the "nerve endings" and this difference is inherent in the myoneural junction. Langley (1905-06) agreed with Elliott as to the place of action of adrenalin but his interpretation as to its mode of action differs from that of Elliott. For Langley the dissimilarit-' of the sjTiaptic

96.

substance must be due to the intrinsic differences in the cells in which the nerve fiber ends rather than to intrinsic differences in the nerve fibers belonging to any one system.

Gruber (1924) noted that adrenalin markedly shor- tened the latent period and the duration of contraction of a skeletal muscular contraction ?.nd induced a greater shortening of the muscle v/hen stimulated, due possibly to increasing the irritability or liberating more available energy or actinpr catalytically on muscle metabolism. Cannon (1929) considered that tiie effect of adrenalin is on the muscle substance. Lutz (1930) interpreted the inhibitory action of adrenalin on the heart of elasmobranch fishes as the response of an unbalanced parasympathetic mechanism in an organ lacking a sympathetic accelerator innervation, and believed the action of adrenalin to be on the nerve endings.

Cannon and 3acq (1931) have reported an adrenalin- like substance which they considered a hormone "as it acts in the same direction as sympathetic im.pulses and in their absence is capable of bringing denervated organs into con- formity with tnose v/hich are innervated, so that the caar- acteristic unified response occurs." because this sub- stance is derived from, stnactures under sympathetic control, when they are influenced by s^nnpa thetic impulses, these investigators suggested calling the substance sympathin, but stated that it is only a provisional nam.e, for as

knowledge of its character increases it may prove really to be adrenalin, developed for local action in smooth viuscle. Wyman and Lutz (1932) have stated that further wor:: is necessary to locate the region of the action of adrenalin in Squalus acanthias .

Unpublished work (1933) of Cannon shov/s that sympathin is not the same as adrenalin. Sympathin may be two sub- stances, one excitatory, the other im.x jpy, called E-sympathin and I-sympathin respectively. Both substances are produced by the nerve endings, ks yet the exact detailed interaction of these substances has not been worked out.

From the results in this investigation it would appear that the action of adrenalin is probably on the muscle fibers rather than the nervous elements, .^s to its mode of action on the muscle fibers it must be taken into consideration that adrenalin is not natural to molluscs. As far as can be determined in the literature the only mention of this substance or a related one, was .nade by Roaf and Nierenstein (1907) who stated that there is a substance in the hypobranchial gland of Pii.ypura papillus which is chemically and physiolo.:';ically allied to adrenalin. Therefore adrenalin when injected into these animals may act as a toxic substance, weak doses exerting a stimulating effect and strone; doses an inhibitory one. On the other hand the excitatory effects produced by weak doses of adrenalin may be due to the fact that it improves the

98.

condition of vertebrate skeletal muscle and also is gen- erally stimulating to invertebrate muscle. This improved condition of the muscle may account for the increase in velocity of a single wave, the velocity of prof^ression and the frequency of waves.

The possibility that adrenalin increases, the irrita- bility of muscle may account .also for the increase of tnese activities when. weak doses are used. It is conceivable that if adrenalin acts on the rayo-neural substance T-ich substance may have different degrees of stajility. i'.ie strena;th of the dose of adrenalin may cause changes in this unstable substance, which in turn causes ~reater or less activity of the factors involved in locomotion.

The proportionality between the rate of creeping and the velocity with which a single wave traverses the foot, or between the freo_uency of .the pedal waves and the velocity of progression is not altered in Helix by the injection of .adrenalin: these facts further support the idea of m.uscle as an "independent effector".

G. Effect of strychnine sulphate on locomotion.

Crozier and Federiehi (1924-25a) have pointed out from their work on the effect of temperature changes on phototropic circus movements of Limax that "a certain type of Dredict^bT lity in ani'^^-i --^ -"T-der 'nor'"'.?!'

natural conditions probably results from dynamic equilibrium

9Q.

thereby obtained between diverse '■^oo'--': r i?^ ^ ^ '^-^ ting for effector control (in the present case, trie creeping mechanisra and that for turninc^, in the ran^^e 14° - 460G.). It follows that unravelling of the elements of conduct necessitates experi;r.entation under diverse abnormal con- ditions favoring individual mechanises of r'=^sPons°."

Th"3e investi£;ators have succeta-o^ 1 : ^ - -a- " :

^^t»..^.i — j.^

these two activities, , orientation and locomotion.

In the experiments on phototropic circus mover.-^nts of ^^^^^ as affected bj changes in temperature tnese authors found that below 15° the amount of circling is determined very largely by speed of creepin*?, v/hil^^ o>^ov9 15O the pedal m-echanism is in secondary place, the turning mechanism becoming the controlling element.

Another method of dissociatir ~ t-ese t^-':^ • -^chanisms was by the injection of strychnine. This substance was found to suppress the phototropic circus movements of Limax msximus (Crozier and Federighi, I'^'^'—^d'r), but no detailed information was obtained concerning the changes in the creeping activity of the foot. Therefore the sub- sequent experiments were performed to study the effect of strycrjiine on the behavior of the pedal waves and to compare them with the normal foot. Small doses of strychnine of definite concentration disturb the functions of synaptic nervous systems, while the same concentration does not disturb the non-s7rnaptic net. By the use of this substance Crozier and Arey (1919) found that in the

100

mo Husk Ghromodorls the general in te "lament rr-3 O "! ''^-o outgrov/ths of the body depend upon a locally contained, peripheral, non-synaptic net-work, but the central nervous system is essentially synaptic. ';/:aile a large proportion of raollusks have the entire nervous system of the non-synaptic type, those v/hich show reversal of in- hibition possess a synaptic nervo^is system.

The ef feet . produced by strychnine, i.e., "reversal of inhibition" is interpreted in various ways by different authorities. In the spinal cord of vertebrates it is usuall3r supposed that tlie strychnine effect is due to the abolition of the inhibitory component of normal coordina- tion, so that the inhibitory effect is transposed into an excitatory one.

. M. and time. Lapicque (1908) have concluded that the effect of strychnine is to bring about a condition of isochronism, i.e. , the chronaxie of the nerve equals that of the muscle fiber and thus the excitation spreads easily from one neurone to another.

It may be argued that the action of strychnine is not due to true reversal but is merely due to a condition of augmented central excitability in -.vhich the excitatory effects produced by stimulation of a mixed efferent nerve conceal the inhibitor:/ effects of the fibers of the sam.e group, i.e., a homogeneous group of inhibitory impulses is not converted into excitatory impulses by strychnine. Magnus and Wolf (1913) and others have observed conditions

101.

which support this interpretation.

Bayliss (1918) has concluded from his experiments on vascomotor reflexes that there may be two independent synapses with which the firr] co'r.rror! notor path connects, unequally sensitive to strychnine, or that the drug acts on some intermediate synaptic membranes on the afferent side, sjTiapses which are not part of the path comraon to the different reflexes.

Bremer and Rylant's (1924, 1925) theory of "reversal" by strychnine is in accordance with that of the Lapicques. They believed that strychnine breaks down excitatory barriers between neurones, thus allowing excitation to spread freely from neurone to neurone throughout the nervous system. It does this by equalizing the chronaxie between various contiguous units which in the nor^.ril un- poisoned nervous system are separated by a bari-ioi- of heterochronism.

Fulton (1926) concluded that true reversal of inJiibi- tion into excitation probably does not occur, ana the apparent reversal is due to the stimulation of excitatorj^ fibers in a mixed nerve, which, cvin'-.- to the increase of excitability produced by strychnine, conceals tiie effect of concomitant inhibition.

Various investigators have recently suggested that too much importance has been given to the question of chronaxie, and have questioned Lapicque's interpretations. Rushton (1930) did not agree with his claim that the

102.

chronaxie of muscle is normally the same as that of the nerve '.vhich supplies it. i-.e strongly supported ^.ous (1907) who maintained that there are two excitable sub- stances in muscle of which one has a very much lar!7er chronaxie than nerve. F-urther, Kushton (1S.32) nas also questioned Lapicque's terminology. Ke (Rushton) stated that "since Lapicque wishes to restrict the n-'i'-rie 'chronaxie' to 'true' chronaxie, it is important to nav^ a nev/ term which can be applied to any strength duration curve 'true' or 'false'. Lucas 'Excitation Time' 'vss ^ic^-r. 'jst this sense and it is proposed for a-or. :ior. . ' ^r.afest (1932) also stated that Lapicque's law of isocnronism is not valid, at least for excitability of the -i---''^ fiber nerve-muscle complex. .-.s agreed w;izr. zr.e views neld by Lucas and Rushton. Lapicque (1932) did not think that Rushton's investigations ^v^^..^ ^^y. lio-ht on

this problem and r.e defenaea r.is position against Rushton. It can be seen that too great importance has been placed on the sub.iect of chronsxie before the various laws claimed for it .lave oeen satisfactorily interpreted and verified.

Although these various theoretical opinions concerning reversal of inhibition exist, it is possible tr_pough pre- cise m.easureraents to obtain data on the behavior of Helix v/hen injected with strychnine. In this w^. - i-^ - be possible to gain information about central nervous activities which could not be obtained by other means.

103.

Although Grozier and Pederighi (1924-25b, 7-221-224) were chiefly concerned with the effect of strychnine on the phototropic movements of Limax maximus and not pri- marily with locomotion, they state that strychnine does not essentially affect locomotion. It was the purpose, therefore, of this investigation to study the character- istic effect of various amounts of str^/chnine on Helix lac tea and Flelix pomatia during locomotion. As far as can be determined no mention of work of this nature can be found in the literature. A study was also made at the same time of the effects of strychnine sulphate, 1 part in 100, on the animals creeping in various planes, i.e., vertically upwards, horizontally (under surface) and horizontally (upper surface).

A series of experiments was first perform.ed to study the effect of strychnine sulphate in the concentration of 1:1000 (1 part in 1000 parts of ninger's solution). Measurements were obtained for the controls and for the in.iected animals during vertical creepir.-. After 0.2 cc. of strychnine sulphate was injected there was either partial or complete retraction of the foot into the shell. The time elapsing after injection before creeping began again varied from fifteen minutes to one hour. It was found that with strychnine sulphate, 1:1000, the creeping was more or less regular and could be compared to normal creeping. The specific results for Helix lactea under these conditions are given in Table XV. Table XVa

104.

TABLE XVa

Effect of doses of str3'-chp-ine sulphate, 1:1000, on the velocity of progression (V), the frequency of v/aves (F), the velocity of single wave (v) and the advance per wave (A).

■^Plus Minus Cases unchanged V 3 4 0

F 3 2 2

v 2 5 0

A 0 3-4

Total number of increases or decreases from the normal animal.

105.

shows that the factors involved durinp- loco-otio- not essentially altered.

Injections of this concentration did not seem to affect the amplitude of the waves, as it -!-3 the same or nearly the same as when the animal was creeping under normal conditions (Table XV, advance per wave).

The effect of stronger concentrations of strychnine were studied for nineteen animals. In the experiments forty-eight series (7150 waves) were obtained for the control animals and forty-four series (6550 waves) for the injected ones v/hen 0.2 cc. strychnine sulphate, 1:100, was injected. Creeping began from fifteen minutes to one hour after the injection. The data obtained from these experiments are given in Table XVI. Reference to Table XVIa shows that strychnine of this concentration did not affect any of the factors involved in locomotion.

Strjrchnine of still greater concentration was used (1:50) but no results could be obtained because after injection when the animal began creeping, sometimes after one or two hours, the creeping was irregular and it was impossible to count the v/aves.

The specific effect of the gravitational pull on tne normal and s trychninized animal during locomotion in three different planes, i.e., vertical, horizontal (under sur- face) &nd horizontal (upper surface) was analyzed. The procedure was to place the animal on a horizontal glass plate which rested on supports about six inches high

106.

TABLE XV.

The effect of strychnine sulphate (1:1000) on vertical ascensions of Helix lactea."^

i\ 0 . 01

Velocity of

Frequency.''

Velocity of

iidvance

experiment

progression

of waves

sin2;le wave

per wave

in mm. per

per minute

in mjn. per

in m]'.

minute

minute

#28 (normal )

9 . 1

45.0

3650

0.20

ffeiy V U • D CC •

s tr ychnine

sulphate

1:1000)

10.0

52.0

7700

0.19

#30(Kormal)

11.2

52.0

7100

0.21

#31(0.5 CC.

strychnine

sulphate

1 : 1000 )

11.8

55.0

8300

0. 21

#32 (Hormal )

4.6

62. 5

6550

0.07

#33(0.5 CC.

"h"r*vp Vtp 1 n ft

sulphate

1:1000)

5.7

62.5

5800

0.09

#57 (Nor-iia 1 )

20. 6

100.0

4500

0.24

#58(0.3 CC.

sulphate

1:1000)

19.0

83.0

4100

0.14

#61 (r ormal )

23 . 1

oo . 0

4oUvJ

r\ or*

#52(0.3 CC.

strychnine

sulphate

1:1000)

8.5

71.0

4600

0.12

#82 (Normal)

16.8

83.0

4600

0.20

#83(0.3 cc.

strychnine

sulphate

1:1000)

14.2

100.0

4500

0.14

#84(norrnal )

27.0

100.0

7100

0.27

#85(0.3 CC.

strychnine

sulphate

1 : 1000 )

22.0

100.0

4100

0.22

Temperature limits were 16.6° to 21.1°G. but the temperature was constant for each experiment.

107.

TABLE XVI

The effect of stryclinine sulphate (1:100) on vertical ascensions of Helix lactea.

No. of

Velocity of

Frequency

Velocity of

^.dvance

exoer inpn t

o I wa V e s

a single wave

per v/£ve

in ram. per

per minute

in mm. per

in rnrn .

LliX ii u. 0 "

minute

4 0

60,6

81.2

3828

0.48

5 e

. 72.1

90.0

4069

0.54

6 0

42.0

72.6

3078

0.36

7 0

39.2

77.4

3772

0.42

8 0

64.8

67.8

3109

0.40

9 •

57.0

79. 2

19 0

57.3

24.9

78.1

0.34

20 A

s"^ n oo . u

<cO . c.

74. 5

0.28

21 0

46.2

29.7

112.0

0.31

p

o^t . o

o D . y

OC . 1

0. 42

23 0

62.8

31.7

90.3

0.40

24 •

55.6

18.9

82.8

0.23

25 0

21.9

96.0

4950

0.29

2d e

19 . 4

70.0

4800

0.27

or? „

27 0

20 . 6

84.2

5150

0.24

28 e

21.3

56.9

4900

0.38

29 0

31.1

91.7

4660

0.31

30 •

31.3

97.0

4920

0.29

31 0

20.8

68.0

5900

0.29

o2 9

27.0

96.6

6175

0.29

33 0

25.0

89.2

5140

0.30

34 •

13.5

69.2

4420

0.20

o o

= control

= injected 0.^1:^1.3] (0.2 cc . of st^^''?"o''inir ^ 9-1 f-p 1:100)

108

TABLE XVI (cont. )

No. of

Velocity of

in mm. per

minute

Frequency u± waves per minute

Velocity of a single wave in mm. per

minute

i.dva"'' ' per in mm.

35 0

36 0

39.6 48.6 .

96.0 54.0

4300

5400

0.44

0.35

37 0

38 •

21,0 13.0

56.0 96.0

4420

0.24 0.18

39 0

40 •

28.8 32.1

88.0 86.9

5010

0.52 0.42

88 0

89 0

29.0 23.1

111.0 68.0

5400 3400

0.26 0.30

90 o

91 •

30.8 11.1

108.0 97.2

4600 4600

0.29 0.10

92 0

93 •

35.0 16.3

98.1 92.1

5500 5160

0.30 0.21

94 0

95 «

35.6 40.0

76.2

ei.o

3500 4000

0.13 0.16

96 o

97 •

13.4 25.0

72.2 98.3

4900 5400

0.19 0.26

o

= control

= injected animal (0.2 cc. of strychnine sulphate 1:100)

109.

TABLE XVIa

Effect of strychnine sulphate, 1:100, on the velocity of progression (V), the f renu.eno^'- of waves (P) the velocity of a single v/ave (v) anu L.ia advance per wave (A).

■^Plus Minus Cases unchRriQ;ed

V 7 10 -2 F 9 10 0

V 9 9 1 A 7 8 4

^ Total number of increases or decreases from the

n or''^/-'' s ""^ i'" 1 •

110

(F±p^, 15). Under the glass plate a mirror was placed at a slope of 45° on a block of wood for the purpose of study- ing the waves when the snail was creeping on the upper surface of a horizontal plane. The methoa or measurement was the same as given above. Data were first obtained for the normal animal when creeping in (1) a vertical plane, (2) in a horizontal plane (under surface) and (3) Horizontal plane (upper surface); after injection of strychinine sul- phate measurements were obtained in the same plane as for the normal animal. The actual scatter was plotted of the various relationships and the slopes of the line were compared.

Pigs. 16, 17 and 18 give typical results and show that in all planes for the s trychninized animal a linear relations'^ip exists between the velocity of a single v/ave and tne velocity of progression, between the frequency of waves and the rate of creeping and between the advance per wave and the rate of creeping. A smaller change in the slope of line is observed for the relationship between the advance per wave and the velocity of progression (Fig. 18) than is noted for the other two relations (Fig. 16 and Fig. 17).

The effect of the gravitational pull on the normal animal during locomotion in the three different planes was observed by comparing the slope of the lines obtained for each relationship. The slope of the lines showing the relationships occurring during vertical creeping was

111.

usually less than for the same relations durlnfr creeping on the horizontal plane, upper or under surface, defer- ence to Table XVII shows that the velocity of a single wave, freouency of waves, the velocity of progression and the advance per wave v/ere usually greater when the animal was creeping in a horizontal plane (upper surface) and creeping vertically upwards than when creeping in a hori- zontal plane (under surface).

In the s trychninized animal not only the change due to strychnine, out also the increase or decrease due to changes in gravitational pull was determined. (Fig. 16, 17 and 18. ) i-xS the gravitational pull is decreased there is usually an increase in the slope of the line describing the various relationships when the animal was creeping on the horizontal plane, upper surface. This increase was less as the gravitational pull was increased (vertical creeping and creeping on a horizontal under surface). However, the results show that when an animal was creeping in each of these two planes tnere was a greater alteration of these factors than when creeping on a horizontal upper surface (Table XVII).

The results in this investigation also show that as the effect of gravity was lessened the factors involved in locomotion were increased, for creeping was increased in the horizontal plane (upper surface) as compared with the other planes. Cole (1925-26) has stated that the stimulus for geotropic orientation and locomotion is the

112.

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tension of the body muscles produced by the downward pull of gravity; this stimulus is received by the propriocep- tors of the muscles. However, if this were the condition we should not expect to get faster creeping when the pull of gravity is lessened; consequently. Cole's theory about locomotion, while it may hold true for orientation, does not seem to sojve the problems involved in locomotion.

The fact that strychnine sulphate in the strongest dose that could be used practically (1:100) did not- essentially affect locomotion may be due to the fact that invertebrates are relatively more insusceptible to strychnine than vertebrates. Also if strychnine acts on synapses, the lack of the usual excitation may be due to a lack of synapses in the intrinsic mechanism of the foot.

Strychnine in this investigation did not abolish the linear proportionality between the velocity of a single wave and the velocity of progression, between the frequency of waves and the velocity of progression and between the advance per wave and the velocity of progression.

115,

Chapter VI. Surnmary and Conclusions

1. The results of these experiments show that in Helix lactea the speed of creeping vertically upward, when not carrying loads, is directly proportional to the velocity of a single pedal wave. The speed of creeping is also directly proportional to the frequency of the waves and to the advance per wave. The proportionality factor is independent of the weight of the animal.

2. The proportionality between the rate of creeping and the velocity with which a single wave traverses the foot, or between the frequency of the pedal waves and the velocity of progressibn, is not altered in normal

or in de-eyed Helix when lifting loads during vertical creeping. Under these conditions the linear relation- ship between the advance per wave and the rate of creeping is not altered. The foot appears to be essentially an independent effector, although under the control of central impulses .

3. The detached foot was observed and kymograms were obtained which showed that the rhythmical action of the foot is greatly interfered with when it is not con- nected with the pedal ganglia. The destruction of the periodic recurrence of waves when the ■oedal 'mnp;lia are separated from the foot suggests that tnis periodicity is determined by the presence of these ganglia.

116

4. Injection of Ringer's solution or raechanical stimulation of the body wall or anterior end of the foot does not affect the factors involved in locomotion.

5. Adrenalin of various concentrations does not

alter the linear proportionality between the velocity of

progression and the velocity of a single wave, between

the frequency of waves and the velocity of progression

and between the advance per wave and the rate of creeping.

These are additional factors which establish tne idea of

the foot

"independent effector" o^/ muscle.

Adrenalin in concentrations of 1:120,000 (0.2 cc. or 0.3 cc. ) produces a stimulating effect on the velocity of progression, frequency of waves, velocity of a single wave and the advance per wave.

An inhibitory effect of adrenalin on invertebrate tissue not previously reported is given. ^-.drenalin in concentrations of 1:10,000, 1:20,000, 1:40,000 and 1:80,000 produces a depressant action on the velocity of progression, frequency of waves, velocity of a single pedal wave, and the advance per wave.

The region of action of the adrenalin is discussed.

6. Strychnine sulphate (1 in 100, 1 in 1000) does not bring about reversal of inhibition in Helix. It does not abolish the linear proportionality between the velocity of a single wave and the velocity of progression, between the frequency of waves and the velocity of progression

and between the advance per wave and the velocity of Pro- gression. These facts are additional evidence that the

117.

foot is essentially an "independent effector".

The effect of , gravitational pull on the normal and s tryclininized animal during locomotion in various planes, i.e., vertically upwards and horizontally (upper surface) is usually to increase the velocity of a single wave, the frequency of waves, the velocity of progression and the advance per wave, while during creeping in horizontal plane (under surface) a decrease in each activity occurs.

7. The purpose of this inves tig;ation was to estab- lish if possible the factors in control of locomotion in Helix. It has been shown that the foot of Helix is pri- marily an independent effector and is secondarily under the control of central impulses. In other words, the intrinsic neuro-muscular mechanism of the foot is the primary factor in locomotion.

118.

Citations

Alvarez, ?i/'.C. and Mahoney, Lucille. The myogenic nature of the rhythmic contractions of the intestine. Am. J. Physiol., 1922, 59, 421-430.

Ascanio, H. and Alvarez, W.G. Studies on the intestinal

muscle of man. Am. J. Physiol. , 1929, £0, 607-610.

Intestinal rhythmicity after death. loid. , 611-616 .

Bayliss, W.W. Principles of General Physiology. London, Longmans, Green & Go. 1918. pp.500.

Biederraan, W. Die locoraotorischen V'/ellen der Schneckensohle . Arch. Ges . Physiol. , 1905, 107, 1-56.

Die Innervation der Schneckensohle. Ibid., 1906, 111, 251-297.

Bohn, G. Des ondes musculaires respiratoires et looomotrices Chez ann^ides et les mollusques. Bull.:_us .ivat. , Paris, 1902, 8, 96-102.

Boyer, P. Action de 1' adrenaline et de sparteine sur le

coeur de I'escargot. G.R.soc.biol. , Paris, 1926,

95, 1244-1247.

Bremer, F. and Rylant, P. Action de la strychnine sur

1 'excitabilite des differents elements de I'arc reflexe. G.R.soc.biol., 1924, £1, 110-113.

Nouvelles recherches sur le mecanisme de 1 'action de la strychnine sur la systbrae nerveux central. G.R.soc.biol,, 1925, 92, 199-202.

Gannon, W.B. Bodily changes' in pain, hunger, fear and rage.

N.Y. and London, D.Appleton & Go., 1929. pp.128.

Gannon, W.B. and Bacq, Z.M. Studies on the conditions of

activity in endocrine organs. Am. J. Physiol. , 1931,

96, 392-412.

Garey, Eben. Studies in the dynamics of histogenesis. J, Gen. Physiol., 1919-20, 2, 357-371. .Anat.Rec, 1920, 1_9, 199-227. J. Gen, Physiol. , 1921, 3, 61-83. Amer , J,Anat, , 1921, 29, 93-115. Amer. J.Physiol. , 1921T22, 58, 182-194. Anat.Rec, 1922-23, 24, 89-96.

Carlson, A.J. The physiology of locomotion in gasteropods. Biol. Bull., 1904-05, 8, 85-92.

I

119

ten Gate, J. Quelques recherches sur la locomotion des Limaces. Arch. N^erl. Physiol. Homme et Animaux, 1923, 8, 377-393.

Contribution a la physiologie du ganglion pedal d'iiplysia limacina. Arch, wierl . Physiol. Hormne et Animaux. 1927-28, 12, 529-537.

Clark, A, J. Comparative physiology of the heart. New York, Macmillan Co., 1927. pp. 62.

Cole, W.H. Geotropisra and muscle tension in Helix. J. Gen. Physiol., 1925-26, 8, 253-263.

Cowie, D.M. and Parson, J. P. and Lashmet, F.H. Studies on the function of the intestinal musculature. Am. J, Physiol., 1929, 88, 363-385.

Crozier, W.J, The rhythmic pulsation of the cloaca of Holo- thurians. Contributions from the Bermuda Biological Station for Research, 1916, III, Ko.43, 297-356,

On the use of the foot in some mollusks. J.Exp.Zool., 1918-19, 27, 359-366.

The analysis of neuro-m.uscular mechanisms in Chiton. J. Gen. Physiol. , 1919-20, 2, 627-634.

"Reversal of Inhibition" by j^tropine in Caterpillars. Biol. Bull., 1922, XLIII, No. 4, 239-245.

On the nervous mechanism of Limax. i^nat.Rec, 1922-23, a, 24_, 398.

Concerning laws of locomotion in gasteropods. Anat. Rec, 1922-23, b, 24, 398.

On the locomotion of the larvae of the slug-moths (Cochlidiidae. ) J.Exp.Zool., 1923-24, 38, 323-329.

On biological oxidations as a function of temperature. J. Gen. Physiol. , 1924-25, 7, 189-216.

Tropisms. J. Gen. Psychol . , 1928, 1, 213-238.

Crozier, W.J. and Arey, L.B. Sensory reactions of Chromodoris zebra. J.Exp.Zool., 1919, 29, 261-310.

Crozier, W,J. and Federighi, H. Phototropic circus movements of Limax as affected by temDera'ture . J. Ge^^ . Physiol. , 1924-25, a, 7, 151-169.

Suppression of phototropic circus movements of Limax by strychnine. J. Gen. Physiol-, 1924-25, b, 7, 221-224.

The locomotion of Limax. J . Gen . Phys iol . , 1924-25, c, 7, 415-419.

120.

Grozier, W.J. and Navez, k.E. The geotropic orientation of gastropods. J. Gen. Psychol. , 1930, 3, 3-37.

Grozier, W.J. and Pilz, G.F. The locomotion of Limax. J. Gen. Physiol. , 1923-24, 6, 711-721.

Grozier, W.J. and Pincus, G. Analysis of the geotropic

orientation of young rats. J. Gen . Physiol. , 1929-30, 13, 57-120.

Grozier, W.J. and Stier, T.J.B. Temperature characteristics

for locomotor activity in tent caterpillars. J. Gen. Physiol., 1925-26, £, 49-54.

Elliott, T.R. The action of adrenalin. J. Physiol. ,1905,32,401-467.

Pulton, J. P. i.Iuscular contraction and the reflex control of movement. Baltimore, Williams and Wilkins Co., 1926. pp.463.

Gruber, G.M. The effect of adrenalin on the duration of the latent, the contraction and the relaxation periods of skeletal m.uscle at rest and undergoing fatigue. J.Pharm.and Exper.Ther. ,1924, . 23, 335-351.

Grundfest, H. Excitability of the single fibre nerve-muscle complex. J.Physiol., 1932, 76, 95-115.

Gunn, J. A. and Underhill, 3.W.F. Experiments on the surviving

mainraalian intestine. Quar. J. Physiol. ,1914,8, 275-296.

Hoagland, H. On the mechanism of an instinct. Thesis (Physiology Library, Harvard University), 1927.

Huxley, Thomas. Science and Education. New York, D.Appleton & Co. , 1897.

Jordan, H. Die Physiologie der Locomotion bei Aplysia limacina. Zeit.f .Biol. , 1901, 41, 196-238.

ti

Jordan, H. and Hirsch, G.Ghr. Ubungen aus der Vergl. Physiol. Berlin, Julius Springer, 1927, 203-263.

Kendall, E.G. Thyroxine. New York. Ghem. Gat. Co. ,1929, 221-230.

Langley, J.I.I. On the reaction of cells and of nerve-endings

to certain poisons, ciiiefly as regards the reaction of striated muscle to nicotine and curare. J.Physiol., 1905-06, 33, 374-413.

Lapicque, L. L' excitabilite' en fonction du temps. Paris.

Les presses univeralitaires de France, 1926. pp.249.

Retrograde polarization, a tneory of systematic errors in measurements of muscular chronaxie through Ringer's Fluid or v/ith large electrodes . J. Physiol. , 1932, 76, 261-281.

121

LaDicoue, L, and Mme. Lapicque. Sur le :necanisnie de la

curarisation. G.R.Soc. Biol. , 1908, 65, 733-755.

Loeb, Jacques. Die Orientierung der Tiere gegen die Scnwer- kraft der Erde (tierischen Geotropismus ) . Zitzber. V/tirzburg.phys . -med.gesellsch. , 1888, Ko.l.

Lucas, K. The excitable substances of amphibian muscle. J.Physiol., 1907-08, 36, 113-135.

Lutz, 3,R. The effect of low oxygen tension on the pulsa- tions of the isolated holothurian cloaca. Biol. Bull., 1930, 58, 74-84.

The effect of adrenalin on the auricle of elasmo- branch fishes. Am. J. Physiol. , 1930, 94, 135-139.

Mackay, M.E. The action of some hormones and hormone-like substances on the circulation of the skate. Gont. Can. Biol. and Fish., N.S., 1^, 19-29.

Magnus, R. Versuche am iiberlebenden D^nndar:a von Saugetieren. i.rch. Ges . Physiol. , 1905, 108, 1-71.

Kflrpers tellung. Berlin. Julius Springer, 1924. pp.29.

Magnus, R. and Wolf, G.G.L. Weitere Mitteilungen ttber den Einfluss der Kopfstellung auf den Glieder tonus . Pfluger's Arch., 1913, 447-461.

Mendel, Lafayette, 3., and Bradley, H.G. Experimental studies on the physiology of molluscs, am. J . Physiol. , 1906-07, 17, 167-176.

Olmstead, J.i.'i.D. Notes on the locomotion of certain Bermudian mollusks. J.Exp.Zool., 1917-18, 24, 233-236.

Parker, G.H. The mechanism of locomotion in gastropods. J.j.iorph., 1911, 22, 155-170.

The locomotion of Chiton. Contributions from the Bermuda Biological Station for Research, 1914, III, no. 31, 1-2.

van Riynberk, G. IV. Sur le mouvement de locomotion de

I'escargot terrestre Helix aspersa. Arch. Keerl. Physiol. , Homme et Animaux, 1918-19, 3, 539-552.

Rose, Maurice. La question des tropismes. Paris, Les Presses universitaires de France, 1929.

Rushton, W.A.K. Excitable substances in the nerve-muscle ' complex. J.Physiol., 1930, 70, 317-337.

122.

Rushton, W.A.K. Lapicque'a canonical strength duration curve. J.Physiol., 1932, 74, 424-440.

Simroth, H. Die Th^tigkeit der vvillldir lichen Muslrulatur unserer Lands chnecken. Zeit.f .will.Zool. , 1378, 50, 166-224.

Sollman, T. A Manual of Pharmacology. Phila., 1928.

Stier, T.J.B. Aspects of the control of animal activities. Thesis, 1928. (Harvard Physiology Library.)

Vl^s, F. Sur les ondes p6dieases des mollusques reptateurs. Gompt. rend. Acad. , 1907, 145, 276-278.

VTfes, F. and Bathellier, J. Sur les lois numeriques des

ondes p^dieuses chez les gas teropoaes . Gompt. rend, ^cad., 1920, Vn, 1085-1086.

Wyman, L.G. and Lutz, B.R. The action of adrenaline and

certain drugs on the isolated holothurian cloaca. J.Exp.Zool., 1950, 57, 441-455.

The effect of adrenaline on the blood pressure of ' the elasmobranch, Soualus acanthias. Biol, Bull., 1952, LXII, 17-22.

123.

Index of Figures

Fig. Al. Diagrams showing: Page 127

a. Helix poniatia, dorsal vie'v

b. " " ventral view 0. " lactea, dorsal view d. " " ventral view

A2. Diagram showing nerve-net in foot of Helix 128

A3. Diagram of Helix showing arranp-ement of ganglia

(cerebral,' pedal and visceral) 129

A4. Schematic representation of the musculature

of the foot of Helix 130

1. Diagram of the method of measurement for

vertical creeping 131

2. The linear relationship between the velocity of a single pedal wave and the rate of creep- ing during vertical ascensions without load 132

3«. The relation between the velocity of progres- sion and the frequency of waves for vertical creeping without load 133

4. Tlie relation between the velocity of progres- sion and the advance per wave for locomotion in a vertical plane without load 134

4a. Comparative graph showing the relation between the velocity of progression and the velocity of a single wave without added load 135

4b. Comparative graph showing relation between

the velocity of progression and the frequency

of waves without added load 136

4c. Comparative graph showing relation between

velocity of creeping and the advance per wave without added load 137

4d. The relation between speed of creeping and velocity of a single wave without added load in Helix pomatia . 138

4e. The relation between velocity of progression and the freouency of waves without added load in Helix poiaatia. 139

4f • Relation between the velocity of progression and the advance per wave without added load in Helix Domatia. 140

124

Page

Fig. 5. The effect of added loads on the relation between frequency of waves and the rate of creeping 141

5a. Actual scatter obtained frotn series of ver- tical ascensions with and without loads showing the relation between the velocity of progression and the frequency of waves 142

5b. Relation between the rate of creeping and the frequency of waves with added loads during vertical ascensions for animal No. 10 143

6. The relation between tension (added loads) and

the rate of creeping - 144

7. The probable error for ti:ie rate of creeping of animal No. 6 as a function of tension (added

loads ) 145

8. Comparison of the normal and de-eyed animal showing the relation between the velocity of a single wave and the rate of creeping 146

8a. iictual scatter obtained from series of vertical ascensions without load for normal and de-eyed animal showing the relation between the velocity of the pedal waves and the rate of creeping 147

9. The relation between the frequency of waves and

the rate of creeping for de-eyed Helix lactea 148

9a. Actual scatter obtained from a series of vertical ascensions without load for normal and de-eyed animal showing the relation between velocity of progression and the frequency of waves 149

10. The relation between the advance per wave and the rate of creeping in the normal and de-eyed animal 150

10a. Actual scatter obtained showing the relation be- tween the velocity of progression and the advance per wave for animal No. 3 v/ith and without eyes carrying no load 151

10b. actual scatter showing relation between the

velocity of pedal waves and the rate of creep- ing for normal and de-eyed animals with added load of 0.7 gram 152

10c. iiCtual scatter showing relation between the velocity of progression and the frequency of waves for normal and de-eyed animal with added load of 0.7 gram 153

125.

Page

Pig.lOd. nctual scatter showing relation between the velocity of progression and the advance per wave for normal and de-eyed animal with added load of 0.7 gram 154

lOe. Comparative graph showing relation between velocity of a single wave and the rate of creeping for de-eyed animal carrying varying loads 155

11. Diagram of the arrangement for recording the behavior of the detached foot 156

11a. Kymograph records of the responses of the and b. detached foot 157

12. The effect of adrenalin on the relation between the velocity of a single pedal wave and the

rate of creeping 158

13. The effect of adrenalin on the relation between

the frequency of waves and the rate of creeping 159

14. The advance per wave as a function of the rate of creeping for the normal and injected animal (adrenalin) 160

15. Diagram showing the method of measurement for creeping in a horizontal plane (upper surface) 161

16. The effect of strychnine on the relation between the velocity of a single pedal wave

and the velocity of progression 162

a. Creeping in vertical plane

b. " " horizontal plane (under

surface )

c. " " " " (upper

surface )

17. Comparison of the normal and injected animal (strychnine sulphate) showing the relation between the frequency of waves and

the velocity of progression 163

a. Creeping in vertical plane

b. " " horizontal plane (under

surface )

c. " " " » (upper

surface )

(

Comparison of the normal and injected animal (strychnine sulphate) showing the relation between the advance per wave and the velocity of progression

a. Creeping in vertical plane

^» " " horizontal plane (under

surface )

c. " " " " (upper

surface )

Figure A 1

Diagrams showing:

^* Helix po!;iatia , dorsal view

b. Helix pomatia, ventral view

c. Plelix lactea , dorsal view

d. Helix lactea, ventral view

FIGURE A I

Figure A 2 ♦

Locomotor nerves of Helix pomatia.

(a) Kerve net in foot.

(b) Central part of nerve system.

1. Ccrebro-pleural connection, cut.

2. Cerebro-pedal connection.

3. Radiating nerves of the pedal ganglion.

4. Cerebral ganglion.

5. Pedal ganglion.

(From Jordan, p. 225, after E. S.chmalz, 1914. )

FIGURE Ai

I

Figure A 3

Diagrams showing arrangement of cerebral ganglion (c), pedal ganglia (P) and visceral

ganglia (v).

(Modified after Richard Hertwig, "A Hanual of Zoology" by Richard Hertwig, trans- lated and edited by J. S. Kingsley, p. 552.)

I

Figure A 4

Schematic representation of the musculature of the foot of Helix. The longitudinal muscle fibers are red, the dorso-ventral black. Pig. 5 is a horizontal, Fig. 6 a vertical longitudinal section. Of the retractor bundles, which are to be considered in Fig. 5 as spread out, only three are represented, rl, r2 and r3. (Taken from Zeitscnrif t f^r wissenschaf tliche Zoologie. 30er Band. Tafel acht. Supplement, Erstes Heft. Leipzig 1878.)

I

Figure 2

The velocity of a single wave is directly proportional to the velocity of progression during vertical ascensions without load. The average deviation of the mean for each point plotted for animal Ho. 7 (Kelix lactea) is as follov/s:

Point 1 1*0

'12 0.25

" 3 0.53

"4 2.9

"5 1.5

Aniraal V/eight of Slope of

animal line

No. 1 6.5 gm. 0.2

II 4 8.6 " 0.36

n 7 7.9 " 0.34

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Figure 3

The frequency of the pedal waves is directly proportional to the velocity of progression, during vertical ascensions without load. This proportionality holds regardles^^ of the weif?ht of the animal. (Helix lactea). (Tahle III).

Animal Weight of Slope of

animal line

IJo. 1 6.5 gm. 0.2

n 7 7.9 " 0.18

" 10 3.8 " -0.2

Figure 4

The advance per wave is proportional to the velocity of progression in vertical creeping without loads. This relationship holds regard- less of the weight of the animal (Helix lactea). (Table III).

Animal Weight of Slope of

animal

line

No. 4

" 10

8.6 gm.

0.3 0.3 0.3

Figure 4A

This comparative r;raph shows that the velocity of progression without loads during vertical ascension is directly proportional .to the velocity with which a single wave tra- verses the foot and that it is independent of the weight of the animal (Helix lactea). Animal Weight of Slope of

animal

line

No. 1

" 3

t» 4

" 5

" 6

w Y

" 8

" 10

" 11

6.5 gi 7.7 ^

8.6 " 7.1 "

6.7 " 7.9 " 8.5 "

3.8 " 5.0 "

:m.

0.2

0.52 0.36 0.48 0.82 0.34 0.52 0.54 0.24

Figure 4B

The frequency of the pedal waves is directly proportional to the velocity of progression during vertical ascensions without loads. This proportionality holds regardless of the weight of the animal (Helix lactea ) .

1 1

z

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Figure 4C

In the analysis of nine animals (Helix lactea) for the relationship between the advance per wave and the velocity of pro- gression the advance per wave is propor- tional to the velocity of progression in vertical upward creeping without loads, and this relationship holds regardless of the weight of the animal (Table III).

Figure 4D

The rate of creeping during vertical ascensions is directly proportional to the velocity with which a single wave courses over the foot v/hen carrying no load. This relationship is illustrated for animals Number 41, 44 and 46. (Helix pomatia. )

-Figure 4E

The rate of creeping is directly proportional to the frequency of pedal waves during vertical ascensions without loads. This relationship is illustrated for animals I'lumber 41, 44 and 46. (Helix por.iatia . )

Figure 4F

The velocity of progression is pro- portional to the advance per wave during vertical ascensions without loads. This is illustrated for animals Number 41, 45 and 46. (Helix pomatia. )

Figure 5

In Helix lactea the linear propor- tionality between rate of creeping (V) and frequency of the pedal waves is not altered when lifting added loads during vertical creeping. This graph shows the individual effect of adding varying loads on animal Number 7.

Slope of line

Without load

0.20

With load of 0.7 gm.

1.2 "

2.0 "

2.5 "

3.0 "

0,20 0.22 0.18 0.42 0.20 0.22 0.22

I

II

I

Figure 5A

The actual scatter obtained from direct observations for a series of runs with and without loads during vertical ascensions. The relationship between the velocity of progression and the frequency of waves has been plotted for animal IJumber 7. (Helix lactea. )

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Figure 5B

This comparative graph shov/s the rela- tion between the rate of creeping and the frequency of waves for animal Number 10 (Kelix lactea ) with added loads during vertical ascensions.

Added load

0.7 gm. 1.2 " 2.0 2.5 " 3.0 3.8 4.8 "

Slope of line

0.18 0.16 0.16 0.14 0.18 0.14

0.28 exhaustion

O CT) 00

1 '

Figure 6

The relation of tension to the rate of creeping is shown from data obtained from ani- mals number 1, 3/4, 6, 7 and 8 during upward vertical creeping (Kelix lactea). The tension is represented by the addition of varying loads, i^., 0.7, 1.2, 2.0, 2.5, 3.0, 3.8, and 4.3 grams. With a great many animals the effect of a load of 2.5 gm. was a decided decrease in the rate of creeping. Usually a slight increase occurred with the addition of 3.0 grams.

I

Figure 7

Figure 7 shows the probable error (calculated according to Bessel's formula) for the rate of creeping with added loads.

Ci>-

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o

(f)

I I L_

(O rj-

9Nld33H3 JO

to

Figure 8

The removal of the eyes does not alter the law of linear proportionality between the velocity of a single wave and the rate of creeping without loads. (Helix lactea. )

Figure 8A

The actual scatter is shovm of data obtained from direct observations of the normal and de-eyed animal (Helix lactea) . The relation between the velocity of the pedal waves and the rate of creeping has been plotted for animal Ko. 3 during vertical ascensions without load.

CO

*

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a u I- 1-

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Figure 9

The linear relationship between rate of creeping and frequency of waves is not altered in the de-eyed animals without load (Helix lactea).

Figure 9 A

The actual scatter is shown of data obtained from direct observations of the normal and de-eyed animal (Helix lactea ) « The relation between the velocity of progression and the frequency of waves has been plotted for animal llo, 3 during vertical ascensions without load.

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m

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Figure 10

The relationship between the rate of creeping and the advance per wave is not altered in the de-eyed animal without load (Helix lactea).

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Figure lOA

The actual scatter is shown of data obtained from direct observations of the norrnal and de-eyed animal (Helix lactea ) . The relation between the velocity of progression and the advance per wave has been plotted for animal No. 3 during vertical ascensions without load.

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Figure lOB

The actual scatter is shown of data obtained from direct observations of the normal and de-eyed animal (Helix lactea ) . The relation between the velocity of the pedal waves and the velocity of progression has been plotted for animal No. 3 during vertical ascensions with added load of 0.7

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Figure lOG

The actual scatter is shown of data obtained fror.i direct observations of the normal and de-eyed animal (Helix l&ctea ) « The rela- tion between the velocity of progression and the frequency of waves has been plotted for animal Ho. 3 during vertical ascensions v/ith added load of 0.7 gin.

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Figure lOD

The actual scatter is given of data obtained from direct observations of the normal and de-eyed animal (Helix lactea ) . The relation between the velocity of progression and the advance per wave has been plotted for animal Ko. 3 during vertical ascensions with added load of 0.7 gm.

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Figure lOE

The effect of added loads is shown on the de-eyed animal (Helix lactea ) « The relation between the velocity of a single wave and the rate of creeping is not altered by added loads during vertical creeping. (See also Table Vila, page 48.)

FIG II. DIAGRAM OF THE A R R AFNGEMEINT FOR RECORDING THE. BEHAVIOR OF THE DETACHED FOOT

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Figure 12.

A comparison of the normal (Helix lactea^)

and injected animal (adrenalin) is given in

this figure which shows the relationship

between the velocity of a single pedal \mve

and the rate of creeping.

Animal #2 Slope of line

o control 0.11 9 injected (adrenalin) 0.21

Animal #14

o control 0.29 ® injected (adrenalin) 0.20

I 1 I I I I I L_

"to VS 50 55 60 6S 70 7S

FIGURE 12. VELDCITY DF PEDAL WAVE,v,

Figure 13

coTiparison of the normal and injected.

animal showing the effect of adrenalin on

the relationship between the rate of creeping

and the frequency of waves.

Animal #2 Slope of line

o = control . 0.38

« = injected (adrenalin) 0.45

Animal #14

o = control 0.22 • = injected 0.24

Figure 14

The relationship between the advance per wave and the rate of creeping is compared with the normal (Helix lactea) and injected animal (adrenalin ) •

Animal #2 Slope of line

0 = control 0.17 » = injected (adrenalin) 0.18

Animal =^14

o = control 0.22 • = injected (adrenalin) 0.18

Figure 16

The relationship betv/een the velocity of a single wave and the velocity of progression is com- pared in the normal and s trychninized animal. This figure shows that strychnine sulphate (1:100) in- creases the slope of the line describing this relationship v/hen the animal is creeping vertically upwards and on the under surface of a horizontal plane. It decreases the slope of this line when creeping takes place on the upper surface of a horizontal plane.

Plane Animal Slope of line

#6

Vertical

o

Control

Injected

( s trychnine )

0.10 0.68

Horizontal (under surface)

o

Control

Injected

( s trychnine )

0.28 0.40

Horizontal (upper surface)

o

Control Injected (strychnine )

1.22 0.87

Figure 17

A comparison of the normal and injected animal (strycnnine sulphate 1:100) shows that strychnine increases the slope of the line describing the relationship betv/een the freoiiency of waves and the velocity of creeping when cne animal is creep- ing vertically upwards, and also when creeping on the upper surface of a horizontal plane, r decrease occurs when the animal is creeping on the under surface of a horizontal plane.

Plane Animal Slope of line

#6

Vertical o = Control 0.68

» = Injected 0.88 (strychnine )

Horizontal

(under surface) o = Control 1.07

» = Injected 0.66 (strychnine)

Horizontal

(upper surface) o = Control 0.42

« = Injected 0.74 (strychnine )

i I I I I I I I I I I 1 1 1 1 LI

35 40 45 50 55 60 65 50 5.5 60 65 W 75 80 85 90 95

FIGURE 17. VELOCITY OF PROGRESSION, V "^^Vsec

Figure IS

A comparison of the normal and injected animal (strychnine sulphate 1:100) shows that strychnine increases the slope of the line describing the relationships between the advance per wave and velocity of pro!3;res3ion when the animal is creep- ing vertically upv/ards . A slight decrease is ob- served Y.'hen creepinging' in a horizontal plane on the upper and lower surface.

Plane Animal Slope of line

Vertical o = Control 0.12

• = Injected 0.26 (strychnine )

Horizontal o = Control 0.32

(under surface) « = Injected 0.30

(strychnine)

Horizontal o = Control 0.22

(upper surface) « = Injected 0.14

t -

8 85 as IQO 105 lio \\5 120 \Z5 i50 05 140 i45 150 155 Q

113 116 IE3 iZ6 13.3 138 143 148 153 158 163 i&8 173 178 183 188 #

HORIZONTAL PLANE UNDER SURFACE

115 12.0 125 130 135 14.0 145 150 155 160 #

VERTICAL PLANE

•

♦6

O = CONTROL

• = INJE.CTED /ANIMAL

1

1

1

1

]

1

1

[STRYCHNJNE SULPH/^Tt)

50 55 60 6.5 70 75 8,0 8.5 9.0 95 lOO 105

FIGURE 18. VELOCITY OF PROGRESSION, V, ""/-sec.

/

Autobiography of the Candidate

Birthplace: Cambridge, Massachusetts

Father's name: John B. Brine, born in Ca:nbridge, i.iass. Mother's name: Mary L. Brine, born in Cambridge, Mass. Education: Grammar School, Public School I\io.26, Kew York City

Hunter High School, New York City

A.B. 1913, Hunter College.

M.Sc. 1915, 'New York University

M.A. 1928, Radcliffe College Positions held: Instructor in the Department of Physiology,

Hunter College, Few York City, from 1913 to 1921. Married: 1921, Mr. John Q. Daly, la\vyer, Boston, Mass.

/ /

//

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