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ON GERMINAL TRANSPLANTATION IN

VERTEBRATES

BY

W. E. CASTLE and JOHN C. PHILLIPwS

WASHINGTON, D. C.
Published by the Carnegie Institution of Washington

1911

CARNEGIE INSTITUTION OF WASHINGTON
Publication No. 144

Paper No. 17 of the Station for Experimental Evolution at
Cold Spring Harbor, New York

Contributions from the Laboratory of Genetics, Bussey Institution,

Harvard University, No. 8.

Copies of this Book
first issued

, 141911

THE CORNMAN PRINTING CO.
CARLISLE, PA.

ON GERMINAL TRANSPLANTATION IN VERTEBRATES.

1. INTRODUCTION.

The scientific results described in this paper were obtained from exper-
iments begun in the Zoological Laboratory of Harvard University and
completed in the Laboratory of Genetics of the Bussey Institution. These
experiments were made possible by a grant from the Carnegie Institution
of Washington to the senior author, for which grateful acknowledgment
is hereby made. The authors desire also to thank Dr. Alexis Carrel, of the
Rockefeller Institute, for valuable suggestions as to operative technique.

The curiosity of zoologists has long been aroused to know whether the
reproductive gland of a vertebrate can be successfully transplanted from
the body of one individual to another; and, if so, whether the gland will
thereafter function in its new environment; and, if it does, whether the
nature of its products will remain unaltered. The fact has repeatedly been
pointed out that experiments of this sort, if successful, should afford a
crucial test of the Lamarckian and the Weismannian views, respectively, of
the relation of the germinal substance to its environment and in particular
to the body.

Our own attention was particularly directed to these questions by the re-
markable results recently described by Guthrie and Magnus, which seemed
to show 'that transplanted ovaries, in a foreign body, liberate products dis-
tinctly influenced in nature by that body. To test the correctness of such
a conclusion the experiments described in this paper were undertaken.
Since it is known that the environment directly influences the nature of
the body, if it can be shown further that the body directly influences the
character of the inheritance through the sexual products, the Lamarckian
principle is established and that of Weismann is disproved. It is therefore
of fundamental importance either to confirm or to disprove the results of
the authors mentioned.

We are unable to confirm, we present evidence which tends to disprove,
the conclusions reached by Guthrie and Magnus. We do not question the
results reported by them, but only the interpretations given by them to that
work.

Every biologist is familiar with the able series of essays in which Weis-
mann showed the physiological distinctness of body and germ-plasm. Many
will recall also the noteworthy experiments of Heape (1890-1897), by which

he showed that influences exerted during gestation do not modify the inher-

1

2 ON GERMINAL TRANSPLANTATION IN VERTEBRATES.

itance. Heape's evidence was this: The fertilized egg of a rabbit of one
variety (for example a long-haired albino) was removed from the oviduct
of the mother before it had become attached to the uterine wall. It was
then transferred immediately to the oviduct of a rabbit of a different vari-
ety (for example a Belgian hare, which is neither white nor long-haired) .
In several cases the transferred egg became attached in its new position
and passed through all the stages of gestation. Young rabbits produced
in this way were both long-haired and albinos like the mother of the eggs,
not like the rabbit which bore the young. The foster-mother, indeed, seems
not to have influenced the inheritance any more than the corn supplied to
cattle determines their breed characters.

Granting all this, a further question remains to be dealt with. The
transferred egg was already full-grown and fertilized. If the transfer had
been made at an earlier stage while the egg was still growing, would the
results have been the same ? Might not the growing egg have lacked that
selective power in assimilation which belongs to the full-grown and fertil-
ized egg ? May not the former be subject to modification by the environ-
ment, even though the latter is not?

This is the question involved in a study of germinal transplantation.
Guthrie believes that he has found evidence of such modification; we ques-
tion the validity of this evidence, on what grounds will presently appear.
On the other hand, we present one clear case of the transplantation of an
undeveloped ovary, which later liberated eggs in the body of a foster-mother,
but in the young so produced no foster-mother influence is detectable. We
therefore question still the existence of foster-mother influence. We
maintain with Weismann not only that modifications of the body are not
handed on to the germ-plasm, but that the character of the body does not
in the least influence the character of the contained germ-plasm, provided
only the body affords a suitable medium within which the germ-plasm may
exist.

2. REVIEW OF THE LITERATURE ON OVARIAN GRAFTING IN ANIMALS

OTHER THAN MAN.

Beginning with the year 1895, a large number of investigators have given
attention to ovarian grafting. The results have often been conflicting. They
represent the work of physiologists, pathologists, biologists, experimental
and clinical surgeons, and lastly of students of heredity. It is impossible
here to give space for the discussion of the entire subject. All that is pro-
posed is a brief review of its more important aspects.

As now understood, the term autoplastic grafting means the transfer of
tissue within the body of the individual, while homoplastic means the trans-
fer of tissue between individuals of the same species. A third term, hetero-
plastic grafting, is used to denote a transfer between two individuals of
different species or genera.

It is proposed to review very briefly the work done in each of these fields .

OVARIAN GRAFTING IN ANIMALS OTHER THAN MAN. 3

Knauer (1896) was the first to report on a series of experiments with ani-
mals. He was led to undertake this work through Chrobak, a surgeon who
had himself tried feeding- ovarian substance to women in whom an artificial
menapause, with its attendant train of symptoms, had been brought about by
operation. Chxobak's results were not conclusive, and he thought surgical
grafting held out more hope of relief in these cases.

In his several papers (1896-1900) Knauer, who worked on rabbits, showed
by a series of twelve autografts that the transplanted ovary persisted in its
new location even up to three years; that its appearance was normal; that
genital atrophy was prevented; and further, that it was possible for animals
so operated upon to bear young. He gives details of his very careful tech-
nique. The ovaries, after the castration, were placed either in the meso-
metrium, on the horn of the uterus, or between the fascia and muscles of the
abdominal wall.

Grigoryeff (1897) confirmed Knauer's work in all its aspects and reported
normal young born from his rabbits after castration and autoplastic removal
of both ovaries.

Ribbert (1897-1898) made careful histological examinations of autoplastic
ovaries, studying the initial process of destruction followed by reconstruc-
tion. As late as 150 days after the operation he found no atrophy.

Fish, Rubinstein, Halban, Herlitzka, Basso, Carmichael, Katsch, Still-
ing, Limon, and others also proved that autoplastic grafting is possible.
From their work one comes to the conclusion that autografts of whole ova-
ries on animals should nearly always be successful, provided the technique
is careful and the ovary not too large or too old. Success does not appear
to depend upon the new position of the ovary. It will grow anywhere
where nourishment is assured, and will even establish itself at times when
merely dropped into the peritoneal cavity. In connection with this, one
must remember the experiment of Lode, who injected the ova of Ascaris
into the abdominal cavity of animals and afterwards recovered these from
the fallopian tubes and uterus.

Among all these workers Arendt raises the only dissenting voice. He
concludes that neither autoplastic nor homoplastic ovarian grafting is pos-
sible. He criticizes Knauer's work and the clinical work of Glass, Morris,
Montprofit, and others, but his conclusions are clearly too sweeping.

We have ourselves obtained several normal young from a rabbit whose
own ovaries were grafted onto the uterine horns. Both the ovaries were
found large and healthy in their new position at the end of nine months.

In studying the results of homoplastic grafting we obtain, however, a very
different picture and are forced to conclude that the success in this group
depends not only on good technique, but also perhaps on the relationship of
the two stocks, and certainly on the intimate chemical tolerance of the

4 ON GERMINAL TRANSPLANTATION IN VERTEBRATES.

opposed tissues. Thus the results are greatly at variance. No doubt the
stock used by some of the workers in this field has been more or less closely
related. The ratio of success in such cases to the degree of relationship
of the opposing tissues has not been worked out, so far as we are aware.

It is necessary to criticize workers in this group on the ground that many-
cases are considered successful if the ovary is found more or less normal
after short intervals — days or weeks. This is no adequate test, as in many
of these cases degeneration is ultimately complete.

Knauer' s results with his thirteen operations of homoplastic grafting were
negative except in two cases, in which the findings were by no means
conclusive. In spite of this Knauer thought that homoplastic grafts were
possible, though difficult.

Fish (1899) experimented upon twenty rabbits with, he says, successful
results. He had hoped to establish the fact that conception is possible
subsequent to homoplastic ovarian transplantation, but in this he was dis-
appointed. His experiment is not given in detail.

Foa (1900-1901) was led to undertake experiments in homoplastic graft-
ing", at the suggestion of Celisea, as a means of settling the controversy
between the Neo-Lamarckians and the Neo-Darwinians. He does not give
details of each experiment, and their number appears to have been small.
His grafts were made onto the original ovarian sites. With this method
one might question whether regenerated tissue could be distinguished from
grafted tissue. His conclusions were that homoplastic grafting was practical,
especially when ovaries of new-born animals were used as material to be
grafted. He thought that such material, planted into older animals, grew
much faster than normally and soon arrived at the growth stage of the
host, and he cites an experiment in support of this view. It is possible,
however, that regeneration of older tissue may have occurred in such cases
rather than accelerated development of introduced tissue. He says also
that ovaries of a new-born animal immediately degenerate when placed in
an older animal whose own ovaries have ceased to function. His findings
inclined him to the belief that there can not be such independence of ger-
minative material as the doctrine of Weismann would have us believe.
Experiments in which he hoped to show the influence of the foster-mother
upon foreign germinative material have not been heard from.

Guthrie (1908) did homoplastic grafting of ovaries in chickens and ob-
tained young from his grafted animals. He concludes that the homoplastic
ovaries function normally and produce young. He thinks that the color
characteristics of the foetuses and of the chicks may be influenced by the
foster-mother. The detailed observations made may be summarized as
follows :

Two pure-bred black single- comb Leghorn and two white single-comb
Leghorn pullets were operated upon, and a third pullet of each sort was

OVARIAN GRAFTING IN ANIMALS OTHER THAN MAN. 5

kept as control. In the operations the ovary of a black pullet was exchanged
with that of a white one. Six months after the operation the birds, which
had now beg'im to lay, were mated, with the following- results in chicks or
foetuses:

1 . The control black hen mated with a black cock produced thirteen black
chicks with light breasts and throats.

2. Black hen B 2, which had received an ovary from a white pullet and
was mated with a white cock, produced nine white chicks and eleven white
ones having black spots on heads, wings, or backs.

3. Black hen B3, which had received an ovary from a white pullet and
was mated with a black cock, produced four ordinary black chicks (with light
under surfaces) and two chicks described as being "black with white legs."

4. The control white hen mated with a white cock produced eighteen
white chicks.

5. White hen W 2, which had received an ovary from a black pullet and
was mated with a white cock, produced three white chicks, one chick white
with black spots, and one ordinary black.

6. White hen W3, which had received an ovary from a black pullet and
was mated with a black cock, produced twelve white chicks spotted with
white on head, wings, or back.

The conclusions to be drawn from these observations will be discussed
elsewhere in this paper.

Magnus (1907) transferred the ovaries from an albino to a black rabbit
with apparent success. The black rabbit was mated with an albino male
five months after the operation and a month later bore two young, one
black and the other an albino. Two months later she died pregnant, and
in the uterus were found two dark-colored embryos and five light-red ones
supposed to be albinos. No ovary was found on one side of the body, but
on the other side was a well-developed and functional ovary bearing corpora
lutea. Magnus supposes that all the embryos produced were derived from
eggs liberated by the transplanted ovary, but in view of our own experience
we are inclined to question this interpretation.

Ten other rabbits similarly grafted by Magnus produced no young, though
three of them gave indications by their sexual activity that they contained
living ovarian tissue.

Among other authors who have reported successful results with homo-
plastic grafting in animals may be mentioned: Schaus, Basso, Mauclaire,
McCone, and Lukaschewitsch.

McCone gives the case of the birth of five well-formed offspring in a rab-
bit from the grafted ovary of another rabbit, but the evidence that the func-
tioning ovary was an introduced ovary is far from complete. Pregnancy
took place four months after complete castration and transplantation from
another member of the same species. The other authors report no young.

6 ON GERMINAL TRANSPLANTATION IN VERTEBRATES.

At variance with these results are the work of the following : Marchese,
Herlitzka, Marshall and Jolly, Burckhardt, and Preobrazhenski, in whose
work degeneration of the implanted foreign ovary was complete or nearly
so after short intervals.

The work on heteroplastic ovarian grafting is small in amount, and the
evidence, taken as a whole, makes it safe to say that persistence of normal
tissue after a few days or weeks is very rare.

Bucura (1907) reports the most favorable results. He transplanted ova-
ries of guinea-pigs into castrated female rabbits. In the first of his three
cases he found after eight weeks the uterus atrophied, but some evidence
of ovarian tissue without follicles. Another rabbit (similarly treated)
was killed after fifty-one days. In this animal there was no genital atro-
phy, and one of the guinea-pig ovaries showed some normal follicles and
a well-developed corpus luteum. The other was completely atrophied.
In the third case, after seventy-seven days, one of the grafted guinea-pig
ovaries was completely resorbed, the other nearly so; while atrophy was
advanced in the rabbit uterus. The author thinks that heteroplastic trans-
plantation is possible.

Basso (1905) reports negative results in grafting ovaries between guinea-
pigs and rabbits.

McCone (1899) cites a case of persistence of the ovaries of a bitch in a
female rabbit for three and a half months with prevention of genital atrophy
in the rabbit. He gives an illustration of this ovarian tissue.

Lukaschewitsch (1901) believes that it is possible to transplant ovaries
from carnivorous to herbivorous animals and vice versa, and that genital
atrophy may be prevented. He obtained no young from animals so oper-
ated upon, and does not describe his operations in detail.

3. REVIEW OF THE LITERATURE ON OVARIAN GRAFTING IN MAN.

It is not surprising when we turn to the clinical experiments to find that
autografts in women are by no means always successful. The material to
be grafted has been adult, and the organ often affected by tumor growth or
cystic and atrophic changes. Small wedges of the organ have been used.

Many authors have advocated conservative surgery in order to avoid the
bad effects of precipitate menapause, but too often cases of this sort have
been reported as successful when the after history was incomplete. It is
safe to say that autografting of ovarian tissue in women has not given
nearly such uniform results as in animals under experimental conditions.

Menstruation has been said to persist in as many as 20 percent of all cases
of ovariotomy. Furthermore, there are numerous cases reported of preg-
nancy after complete ovariotomj\ Doran (1902) cites three such cases. In
one of these, observed by himself, a definite focus of extra-ovarian tissue
was found at a second operation in the ovarian ligament.

OVARIAN GRAFTING IN MAN. 7

Dauber (1905) shows how difficult it is, especially in disease, to assert
with confidence that all ovarian tissue has been removed.

Dudley (1900) cites several cases of pregnancy after ovariotomy.

Croom (1905) reports a case of homoplastic ovarian graft, with four years
later the birth of a living- child. Obviously it is impossible to verify such
cases as this, and they must always remain open to doubt unless the child in
question shows some strongly inherited and dominant characteristic which
was possessed by the individual who supplied the graft, but not by the
foster-mother or by the father. Such conditions are not likely often to be
realized in clinical experience, hence our principal reliance for determin-
ation of the theoretical questions involved must be animal experimentation.

Morris (1906), who had done much successful clinical work on autograft -
ing, besides some early experimental work, reported the birth of a living
child in a woman into whom, after careful castration with the Tuffier angio-
tribe, he had grafted wedges of ovary from another woman. The castrated
ovaries of the woman who was grafted were cirrhotic, and no formed
Graafian vesicles were found. It was assumed that the introduced ovarian
tissue was responsible for the result reported; but Lucas-Championniere,
criticizing this case, has expressed the view that it can be given another
interpretation. He cites as an example a patient of his own, in whom, after
the most careful castration with the angiotribe for a case of dysmenorrhea,
the symptoms still persisted. A second operation on this patient, over a
year later, revealed an important ovarian fragment at the uterine horn.

Glass (1899) gives a convincing case of homoplastic operation in a woman
who two years before had had ovariotomy performed. She was suffering
from all the symptoms of an artificial menapause, but a graft from another
woman relieved her. The relief appears to have been permanent. No
children are reported.

Mauclaire (1900) collected thirteen cases of homoplastic operation in
women. In six of these cases menstruation returned, and in two cases it
was resumed temporarily. The persistence and functioning of the intro-
duced tissue is, however, in no case established beyond question. On the
whole, Mauclaire regards ovarian transplantation as of doubtful clinical
value.

Sauve (1910) sums up in a very clear and direct way all the surgical evi"
dence at hand. His statistics show that return of menstruation is no more
common after ovarian grafting than after ovarian castration. There are,
he thinks, only two cases of successful homoplastic grafting in women.
The first case is that reported by Morris, in which a woman with infantile
ovaries became normal and had regular menstruation after the transfer of
ovarian tissue from another woman; and the second case is that of Pankow,
which is supported by definite histological evidence after a period of three
years.

8 ON GERMINAL TRANSPLANTATION IN VERTEBRATES.

4. NEW OBSERVATIONS ON OVARIAN GRAFTING.

In this paper we shall report upon seventy-four cases of homoplastic
ovarian grafting in guinea-pigs and seventeen similar cases in rabbits.
The purpose of the experiments represented by these cases has been stated
in the introduction.

The operative technique in the guinea-pig was as follows: The hair of
the female to be grafted was removed from each side at the costal border.
She was then anesthetized and was castrated through an incision on each
side, about an inch long, at the costal border and usually a little ventral to
the ovarian site. The ovary was drawn into the field with blunt forceps
and lifted with a small, double eye-hook, after having been carefully shelled
out from the tube and mesentery. The organ was then cut away, care
being taken not to damage the tube. The animal from which grafts were
to be taken was then quickly killed and its ovaries removed, but with a small
bit of mesentery attached. The ovaries themselves did not come into con-
tact with any instrument. They were attached by means of very fine silk
(00 untwisted into thirds) and a header's needle, to various sites in the
peritoneal cavity. Some even were merely dropped into the abdominal
cavity. The muscles and skin were closed separately. At first two opera-
tions were made for each animal; but this was soon found to be unneces-
sary. The mortality was very slight. Out of the seventy-four cases six
died as the immediate result of the operation; four of these were cases in
which a ventral incision was tried.

To sum up the result of the entire series, only one* grafted animal had
young from her grafted tissue; gTafted ovaries functioned in six other cases,
but did not produce young. Ten animals regenerated their own ovaries,
and three of these had young. Forty-two showed post-mortem complete
atrophy of the genital tract and absence of ovarian tissue. The remainder
comprises fifteen cases in which results were not fully determined.

It is thought worth while to give an account of all the groups, because,
though only one* case, Group I, bears on the problem before us, the series
may have some physiological significance on account of its very consider-
able size, and may throw a side-light upon what criteria are necessary in
such work.

Group I.

This group includes homoplastic transplantation of an ovary resulting
in the birth of young derived from grafted tissue. On January 6, 1909,
the left ovary was removed from an albino guinea-pig, No. 27 (fig. 2, pi. l),
then about 5 months old, and the ovary of a pure black guinea-pig (compare
fig. 1, pi. l), about a month, old was fastened near the tip of the uterine
horn, distant a centimeter or more from the site of the ovary removed. One
week later, January 13, a second operation was performed, in which the
right ovary of the albino was removed, and as a graft was introduced the

A second case, from a new series of experiments, has just been observed (January,
1911); see Postscript, p. 10.

NEW OBSERVATIONS ON OVARIAN GRAFTING. 9

ovary of a second young" black guinea-pig-, of like age with the first but of
different ancestry. After the albino had fully recovered from the second
operation, she was placed with an albino male, No. 654 (fig. 3, pi. l) with
which she remained until her death about a year later.

On the 23d of July, 198 days after the first operation, she gave birth to
two female young-. One was black but bore a few red hairs. A photograph
of this animal (No. 1970) at the age of three or four months is shown in
fig. 6, pi. 2. The other young one (No. 1969) was likewise black, but had
some red upon it, and its right forefoot was white. (See fig. 5, pi. 2.)

On October 15 the grafted albino bore a third black young one, a male
which, like those previously born, had a few red hairs interspersed with the
black. A photograph of this animal is shown in fig. 4, pi. 2.

On January 11, 1910, the grafted albino was observed to be pregnant for
the third time, and this time she was very large. Unfortunately, on Feb-
ruary 2, she died of pneumonia with three full-grown male young in utero.
The skins of these animals were saved and a photograph of them is shown
in figs. 7-9, pi. 2. Like the other three young they were black, but with
a few red hairs among the black ones. They bore no white hairs.

An autopsy made one hour after the death of the mother showed on the
left side a distinct ovarian mass about a centimeter from the coiled part of
the oviduct; that is, approximately in the position where the graft from the
pure black guinea-pig was fastened at the first operation. On the right
side the mesentery of the oviduct was adherent to the body-wall where
an incision had been made at the second operation, and a small amount of
tissue, regarded as possibly ovarian, was there observed. Both this and
the ovarian tissue from the left side were preserved in Gilson's fluid for
sectioning and microscopic examination. The tissue from the left side was
found to contain numerous large egg-follicles, some already well advanced
containing a lymph space; in addition a number of corpora lutea were
observed. On the right side was found a small amount of undoubted ova-
rian tissue, with one well -advanced egg-follicle, but the whole apparently
was strongly encapsulated, so that no eggs could be discharged even if they
came to maturity.

It is interesting to note that both grafts persisted, though taken from
different animals and transferred at different times. This result suggests
a possible susceptibility on the part of the animal grafted.

Female 1970 (fig. 6, pi. 2) daughter of the grafted albino, was mated
with the albino male, her father (fig. 3, pi. l), and bore three young, two
of which were albinos and one black with some red hairs. If female 1970
had been the daughter of a pure-black mother, instead of a grafted albino,
we should have expected her to produce an equality of black and of albino
young. The observed result was the nearest possible numerical agree-
ment with this expectation.

A control mating of the albino male, 654, was made with a female of
pure black stock. As a result there were produced two litters of young,

10 ON GERMINAL TRANSPLANTATION IN VERTEBRATES.

including- five individuals, all black, but with red hairs interspersed. This
result shows that the red hairs found on the six young of the grafted albino
were due, not to foster-mother influence of the grafted albino, but to influ-
ence of the male parent. The young of the grafted mother were exactly
such in color as the black guinea-pig which furnished the graft herself
might have been expected to bear had she been mated with male 654 instead
of being sacrificed to furnish the graft. The white foot borne by one of the
young forms no exception to this statement. Spotting characterized the race
of guinea-pigs from which the father came. He was himself born in a litter
which contained spotted young, whereas neither the pure-bred black race
that furnished the graft nor the albino race that received it was character-
ized by spotting.

Inasmuch as the offspring of albino parents are invariably albinos, it is
certain that the six pigmented offspring of the grafted female were all
derived from ova furnished by the introduced ovarian tissue taken from a
black guinea-pig. This tissue was introduced while the contained ova were
still quite immature, and it persisted in its new environment for nearly a
year before the eggs were liberated which produced the last litter of three
young. These young, like the earlier litters, gave no indication of foster-
mother influence in their coloration.

The conclusion is forced upon us that the egg-cell during its growth does
not change in germinal constitution. Its growth is like the growth of a
parasite or of a wholly independent organism : what it takes up serves as
food; this is not incorporated merely in the growing organism; it is made
over into the same kind of living substance as composes the assimilating

organism .

POSTSCRIPT.

While this paper was in press a second case, from a new series of experi-
ments, belonging in Group I came to light.

An albino female, No. 2475, born on the 22d of March, 1910, at 75 days of
age was operated upon, being, doubly castrated and receiving the ovaries
from a half-sister, No. 2507, a brown-eyed cream animal, 17 days younger
than herself. She was then mated with an albino male, her half-brother,
No. 2402. On October 20, 136 days after the operation, she gave birth to
an albino young one. We considered this an indication that the castration
had been imperfect, but allowed the animal to breed again. Much to our
surprise, 73 days later, on January 1, 1911, she gave birth to two young,
one of which was a brown-eyed cream like the animal which furnished the
graft. The other young was an albino.

The albino young borne by this grafted animal can not properly be
regarded as evidence of somatic influence on the introduced graft. For
albinism occurred as a recessive character in the particular brown-eyed
cream stock used, as is evident from the fact already stated that the colored
animal which furnished t' e graft and the albino which received it were
half-sisters. The character of the young obtained and their numerical pro-
portions are exactly such as the colored animal would herself have been
expected to give, had she not been sacrificed to furnish the grafts, but had
been mated with the albino male.

CASTLE

PLATE 1

1. A Young Black Guinea-Pig, about three weeks old. Ovaries taken from two

animals like this were transplanted into the albino shown in Fig. 2.

2. An Albino Female Guinea-Pig, No. 27. Its ovaries were removed and in their place

were introduced ovaries from two young black guinea-pigs. Compare Fig. I.

3. An Albino Male Guinea-Pig, No. 654, with which was mated the albino shown

in Fig. 2.

CASTLE

PLATE 2

4, 5, and 6. A Group of Three Young, produced by the pair of albinos shown in
Figs. 2 and 3. 4, is female 1969, 5, is female 1970, 6, is the male born on
October 15. All were about half grown when photographed.

7, 8, and 9. Skins of three young found in utero at the autopsy of female No. 27,
Fig. 2. The sire of these young also was male No. 654, Fig. 3.

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12 ON GERMINAL TRANSPLANTATION IN VERTEBRATES.

left incision. Ten and a half months later the animal was killed. The
uterus was well nourished. There was no tissue at the sites of the original
ovaries, but attached to the border of the greater omentum was a typical
ovarian mass which showed microscopically plenty of ovarian tissue and
at least one large ovarian follicle. This case is interesting as showing that
presence of ovarian tissue in immediate contact with the genital ducts
is not necessary to prevent atrophy of those ducts. An ovary attached to
the digestive tract (or doubtless anywhere else in the body) serves equally
well to prevent genital atrophy. Some substance liberated by the ovary
and circulating in the blood is doubtless the active agent in maintaining
the full development of the uterus and the recurrence of rut.

In guinea-pig No. 73, one year old, the grafts were placed, one attached
to the right body-wall, the other merely dropped into the abdominal cavity.
Nine and a half months later the animal was killed. There was little or
no genital atrophy, and at the site of the graft on the body- wall a small
mass was found which was identified by microscopic examination as the
oviduct of the graft, with a small amount of degenerating ovarian tissue,
without follicles. There was no sign of regenerated tissue at the ovarian
sites.

The above six cases show that normal ovarian tissue persisted in its new
environment for periods from four months to ten and a half months. They
demonstrate that the position of a homoplastic graft may be varied to any
place on the peritoneal surface; and that the grafted ovary may attach and
nourish itself without the aid of sutures.

The failure of these foreign ovaries to bring about pregnancy inclines
one to the idea that the ovum when mature and liberated may be more
subject to resorption during its migration to the uterus than the more prim-
itive tissue from which it was discharged. It is also well to remember that
Graafian follicles have been found to be less in number and larger in size
in grafted tissue than in normal tissue (Carmichael, Sauve). Further,
in the case of an ovary removed from its normal position, the chances are
necessarily greatly diminished that the egg when liberated will find its way
into the oviduct. In an ovarian operation there is also great danger that
the open end of the oviduct will become permanently closed in consequence
of injury. On the whole it is not surprising that foreign ovaries often
persist without bringing about pregnancy.

Group III.

This group includes ten cases in which, in spite of a supposed perfect
castration, the animal regenerated ovarian tissue at the original site. Three
of these animals bore young. In all ten cases regenerated ovaries were
found post mortem by inspection and microscopic examination. The color-
character of the young indicated that they had probably come from regen-
erated tissue, and the autopsy in all cases confirmed this view.

NEW OBSERVATIONS ON OVARIAN GRAFTING. 13

The three cases in which young' were born are as follows:

Guinea-pig- No. 77, blue, age five months, underwent a double ovarian
operation in which grafts were taken from a pale cinnamon agouti guinea-
pig, age one week. Both grafts were placed on the right side at the uterine
horn. Six months later she bore by a white male, No. 654 (fig. 3, pi. l),
a blue female young one . She was killed eight months after the operation
and ovarian tissue was demonstrated at the right ovarian site, while the
graft had completely disappeared.

Guinea-pig No 24, an albino, age four weeks, was grafted from a black
animal one day old. One graft was stitched to the body- wall on each side.
Two and a half months later she was noted p regnant, and at three months
she had, by male 1078 (a brown-eyed yellow), two young, both black-eyed
yellow. Her post mortem two days later showed indications of ovarian
tissue at both ovarian sites, but the microscopic examination showed that
ovarian tissue was confined to the right ovarian site. There was nothing-
left of either graft.

Guinea-pig No. 1907, black, age three weeks, whose parents were one
black , the other cinnamon agouti marked with cream, was grafted with
ovaries from a cinnamon agouti, age five days. One graft was dropped
into the abdominal cavity, and one was attached to the body-wall on the
right side near the incision. Five months later she bore, when mated to male
No. 654, fig. 3, a young black male with red brindling. The post-mortem
showed at the right body- wall a pinkish mass 1.5 mm. in diameter, which
was preserved. Both ovarian sites were also saved for microscopic exam-
ination. The peritoneal cavity was negative. The microscope showed
that the mass on the body-wall consisted of much fibrous and connective
tissue penetrated by blood-vessels, the remains of the degenerated graft.
The right ovarian site showed normal ovarian tissue with Graafian follicles
present.

The other seven cases, in which no young were produced, but in which
the presence of regenerated ovarian tissue was demonstrated by the micro-
scope, need not be given in detail. A well-developed condition of the uterus
was observed in all.

Group IV.

In this group are included those cases, forty-three in all, which showed,
by complete atrophy of the genital tract, absence of all ovarian tissue. Care-
ful microscopic inspection was made in all cases, but nothing resembling-
ovarian tissue was in any case observed. Four of these animals died from
two to nine months after the operation ; the remainder were killed at periods
ranging from four months to one year afterward . The average age of the
lot at the time of the operation was eight weeks. The individual age of
the transplanted ovaries varied from one day to two months, while the
average age was three weeks.

14 ON GERMINAL TRANSPLANTATION IN VERTEBRATES.

Group V.
In this group are placed fifteen cases in which the grafted animals died
or were lost without any autopsy being- made upon them. Only six died
as the immediate result of the operation, four of the six being cases in which
a ventral route to the ovaries was attempted.

The results obtained by us from ovarian grafting in rabbits are largely
negative. Seventeen animals have been operated upon.

One rabbit, in which an autograft was made onto the uterine horn on each
side, gave birth four months later to two young. Nine months later a
second operation showed both these autografts functioning ; they were
removed and ovaries from another rabbit were placed in their stead; atrophy
of the external genitalia soon resulted and the animal was a few months
later discarded.*

Of the other homoplastic cases, two showed regenerated tissue at the
original ovarian site, one of these having had young. Of the remainder,
six died or were lost, while three showed complete atrophy of the external
genitalia and three more are still alive, manifesting normal sexual instincts
but having borne as yet no young.

On the whole the results obtained from rabbits are very similar to those
obtained from guinea-pigs. Autografting is evidently not difficult, and
may be followed by the birth of young. In heteroplastic grafting, regener-
ation of ovarian tissue at the original site is a commoner event than growth
of the introduced tissue, the latter result not having been certainly obtained
in our experiments with rabbits, though the former has been obtained in
two cases.

5. CRITICAL EXAMINATION OF THE OBSERVATIONS OF GUTHRIE AND MAGNUS ON

SUPPOSED FOSTER-MOTHER INFLUENCE.

A cursory reading of Guthrie's evidence may well lead one to accept
without question his conclusions: (l) that in his fowls the transplanted
ovaries have functioned, and (2) that the foster-mother has influenced the
character of the young; but critical analysis of the evidence fails to sub-
stantiate either conclusion. Since the ovaries exchanged were transplanted
to the original ovarian site, it would have been impossible by autopsy to
ascertain whether the ovary present was transplanted or regenerated
tissue. But regeneration is a much commoner result of such operations
than successful transplantation, as our results with guinea-pigs show; and
in the absence of other evidence the interpretation of the functioning ovary
as a regenerated ovary should be preferred. The only other evidence we
have is that derived from the character of the offspring.

*Simple inspection of the external genitalia appears to be sufficient in rabbits to show
whether or not the uterus is functioning normally.

EXAMINATION OF OBSERVATIONS OF GUTHRIE AND MAGNUS. 15

Since in Guthrie's experiments the character of the germ-cells furnished
by the mother is in question, we may conveniently group the experiments
according- to the character of the male used.

Series I. Rooster Black.

Black hen Bl, control, mated with the black rooster, had 13 black chicks
with "grayish-yellow breasts and throats, and frequently the under sur-
faces of the tips of the wings were light colored as well, but the plumage on
the entire dorsal surface was always solid black."

Black hen B 3 and white hen W 3 had their ovaries exchanged and were
later bred to "the black rooster." Black hen B3 produced six black
chicks, four apparently normal, but two " with white legs." We are not
told whether this whiteness was of down or final plumage, but the state-
ment is added: "In regard to the chicks from this hen described as ordi-
nary black, some doubt exists as to whether the ventral light-colored area
described for normal black chicks was not lighter and greater in extent in
all cases than in the normal chicks." Since this ventral lightness is of
down only, not of later plumage, we are led to infer that the description
of the abnormal black chicks was also based on the down colors. And
since, further, white chicks are not white in the down, but blacks chicks are
light be\ow in the down, it seems that Guthrie's real meaning is that the
two abnormal chicks had light legs (not white legs) in the down. But
this does not indicate whether the adult plumage in that region was to
be black or white. If this reasoning is correct, then the only difference
between these two chicks and ordinary black chicks is that they perhaps
had more extensive light ventral areas in the down plumage, the light area
extending onto the legs. This has no relation whatever to the plumage
condition of the white race from which the graft was taken and can not
fairly be assigned to that source.

If the six chicks were produced from regenerated ovary (i. e. , from germ-
cells of the black hen not removed at the operation) we should expect them
all to be black, as they were. If they were produced from transplanted
ovarian tissue without foster-mother influence, we should expect them all
to be white, which they were not. For the white of Leghorn fowls, accord-
ing to Bateson and Davenport, behaves as a dominant character in crosses
with black, cross-breds being white or white spotted sparingly with black.
Unless, therefore, we grant that these six chicks came from regenerated
ovarian tissue of the black mother herself, we are forced to assume that the
ovarian tissue transplanted from the white hen completely changed its char-
acter from white to black within the black hen. It is a hypothesis at least
equally plausible to suppose that no cells were changed in character, but a
few were overlooked in the castration.

White hen W 3 , having received the ovary from black hen B 3 , was also
mated with ' 'the black rooster. ' ' She produced twelve ' 'white chicks with
black spots on the dorsal surface of the head, neck, wings, back, or on the

16 ON GERMINAL TRANSPLANTATION IN VERTEBRATES.

tail." This is such a result as we should expect had the hen not been oper-
ated upon at all. It is fully accounted for if we assume that the ova which
produced these twelve chicks came from regenerated ovarian tissue of the
white mother herself. On the other hand, if the transplanted ovary had
produced the chicks without any foster-mother influence, the chicks should
have been all black with light ventral surfaces, which they were not. Again
the regeneration hypothesis presents far less difficulty than that of a suc-
cessful transplantation.

Series II. Rooster White.

White hen W 1 control, mated with the white rooster, had eighteen chicks
"pure white to light buff when hatched." White hen W2 and black hen
B 2 had their ovaries exchanged, and were later bred to the white rooster.

Black hen B 2 produced nine white chicks and 11 white spotted sparingly,
with black. Such a result as this is the usual consequence of a cross
between black fowls and white ones; that is, it is what might have been
expected had the black hen never been operated upon. And, on the other
hand, if the functional ova came from the transplanted ovary, the expectation
would not be materially different, though the breeding capacity of the young
would be different in the two cases. Unfortunately this was not tested.

The white hen W2, which had received the black ovary from B 2, pro-
duced five chicks. Three were white, one white spotted with black, and
one black. A white hen not operated upon might be capable of producing
all three sorts when mated with a white rooster of a similar character;
that is, both parents might be Mendelian heterozygotes bearing black as
a recessive character, in which case one-quarter of the young should be
black. Now, the numerical result is not at variance with such an interpre-
tation. But if the same white rooster was used in this mating as with B 2,
it is surprising that no black young were produced by B 2. Guthrie does
not expressly state that the same rooster was used in both cases, but we
assume this to have been the case from his use of the expression "the white
rooster." If different males were used in the two cases, one may well have
been homozygous, the other heterozygous. In that case the black chick
as well as the white and the spotted ones may have come from regenerated,
not from transplanted tissue. But if the same white rooster was used in
both cases, it is still equally difficult to account for the black chick as a
product of a transplanted germ-cell or of regenerated ovarian tissue of the
white mother. For unless the white male was heterozygous in black we
should expect no black young to be produced even were every egg which he
fertilized produced by a pure-black hen, instead of by a white hen possibly
carrying borrowed black germ-cells.

Control matings of the white rooster with normal black hens, or of the
black chick when it became adult, would have cleared up the case, but no
control crosses were made by Guthrie. The control mating of "the white
rooster" with a normal white hen Wl was not a sufficient control of his

EXAMINATION OF OBSERVATIONS OF GUTHRIE AND MAGNUS. 17

breeding- capacity. It does not prove him to have been homozygous in
white. If W 1 was homozygous, only white chicks would be expected, what-
ever the character of the white rooster.

We have now examined in detail the two pairs of transplantation exper-
iments which form the entire basis of Guthrie 'e assumption that the foster-
mother may in a case of homoplastic transplantation affect the character of
the offspring. In no one of the four cases does the hypothesis that the
transplanted ovary functioned offer less difficulty than the hypothesis that
only regenerated ovarian tissue functioned. In two of the four cases the
advantage is overwhelmingly with the latter hypothesis. That ovarian
tissue might easily be left behind in exchanging ovaries between two ani-
mals Guthrie frankly admits in a recent paper (1910), citing experiences
of his own to show just how this might come about. He still maintains,
however, his belief in foster-mother influence, because, we believe, of a fail-
ure to grasp fully the laws of inheritance of the character which he used as
a criterion. A similar failure is shown by the comment which he makes
in Science (1909) upon the case which we have more fully described in this
paper (Group I) . Referring to our failure to detect foster-mother influence
in the young borne by female 27, he says : "Had the operated pig been
bred to a male of the same strain as the pig from which the engrafted ovary
was obtained * * * characteristics in the offspring indicative of such
influence might have been obtained." Now, suppose the operated pig had
been so bred, what result might have been expected? Exactly that which
was obtained from the mating with an albino, only we should have been left
in uncertainty, precisely as in Guthrie's own experiments, as to whether
the pigmentation of the offspring was due to maternal or paternal influence.
By the'mating with an albino male all such uncertainty was eliminated,
since it was rendered sure that if the offspring were pigmented they could
have derived this character from no other source than the transplanted
ovary of the little black guinea-pig.

Our case does not prove that foster-mother influence is impossible. No
such claim is made for it. But our observations do show that evidence such
as Guthrie has presented is wholly without value in establishing foster-
mother influence, and that in one specific case, the first critical case on rec-
ord, we believe, no foster-mother influence is detectable.

In view of our own experiments on guinea-pigs, in which the source of
the tissue which liberated the ova is fully established and in which no foster-
mother influence is detectable, we may fairly ask that the experiments with
fowls be repeated on adequately controlled material before we accept the
interpretation which Guthrie has given to his results.*

*While this paper was in press our attention was called to the fact that Dr. C. B.
Davenport (1910, Proc. Soc. Exp. Biol. Med., vol. vn, p. 168) had indeed repeated Guth-
rie's experiment with fowls, on material adequately controlled and of known pedigree.
The result was that in every case castration was incomplete and ovarian regeneration
occurred, leading to the production of young showing no influence of the introduced
graft.

18 ON GERMINAL TRANSPLANTATION IN VERTEBRATES.

The work of Magnus demands no extended discussion. Into a black
rabbit were introduced the ovaries of a white one. The grafted black rabbit
was then mated with a white male rabbit. Two litters of young resulted
which consisted in part of black animals and in part of white ones. There
is no question of an intermediate or modified character in the young, such as
Guthrie would have us believe occurred in his experiments. If any foster-
mother influence was exerted at all, it was such as changed the character
of germ-cells completely from the albino to the fully pigmented condition.
But this supposed influence manifested itself in part only of the germ-cells
subjected to the new somatic environment; the others were unaffected.

An alternative interpretation is plainly possible. The ovary observed
at the autopsy of the grafted black rabbit may have been regenerated from
fragments of ovarian tissue not removed by castration. We do not under-
stand that there was anything in the observed position of the organ decisive
either for or against such an interpretation.

It is known from studies of Hurst and of Castle that the albino condition
in rabbits is a Mendelian recessive to black. The grafted black rabbit may
have been a heterozygous black, containing albinism as a recessive char-
acter. In that case, if the black rabbit had not been operated upon, but had
been mated with an albino male, we should have expected that young would
be produced half of which were black, half albinos. This is substantially
the result observed. Accordingly, so far as the character of the young is
concerned, there is no reason to think that they were produced from intro-
duced ovarian tissue rather than from regenerated ovarian tissue of the
black rabbit herself.

But if we adopt the interpretation that the observed ovary was really an
introduced ovary, we encounter this difficulty. If introduced egg-cells
through somatic influences were changed in character so that they produced
black young instead of white ones, why were not a// the egg-cells so affected?
Were they not all subject to the same somatic influences? On the whole,
therefore, we meet fewer difficulties by interpreting the observed ovary as
a regenerated ovary rather than as a transplanted ovary.

6. A REVIEW OF THE LITERATURE ON GRAFTING OF TESTICLES.

The work on testicle grafting is less voluminous than that on ovarian
grafting. In a general way some of the more uncritical of the early inves-
tigators reported results of a positive nature; but later work along the same
lines gives results of an entirely different character.

Berthold in 1849 interchanged testicles between two cocks after castrating
both animals. The organs, one in each animal, were not sutured. Five
months later they were found grown against the colon in both cases, and
full of active spermatozoa. The cocks retained their sexual instincts.

Mantigazza (i860), experimenting on frogs, found living testicular tissue
in grafted animals after a period of 70 days. He thought that the tissue
preserved its structure. Bizzozero (1868) confirmed these results.

GRAFTING OF TESTICLES. 19

Cheveau (1890) in a brief note described autoplastic testicular transplan-
tation, apparently of adult tissue, in sheep. He obtained nourishment of
the exterior of the testicle, but concludes that the operation is futile if the
testicle is really detached from its own blood-supply.

A. Lode (1891) reported the results of experiments on cocks which were
castrated, their organs being- transplanted into subcutaneous connective tis-
sue. One cock eight months after operation revealed a tumor of bean size
under the skin. This appeared vascularized and contained living sper-
matozoa. Another cock, in which, after being caponized by a professional
caponizer, the testicles were thrown back into the body cavity, showed a
bean-sized testis attached to the peritoneum at the site of the old wound,
containing living spermatozoa. There were also numerous spermatozoa
contained in little testicles in various parts of the body cavity, which were
regenerated bits of the testicles, crushed in the process of caponizing. He
states that he always found testes remaining in capons, no matter how care-
fully castrated. This work therefore would allow one to make a somewhat
different interpretation of Berthold's work than he himself made: the sup-
posedly transplanted testicles may have been regenerated from fragments
never removed.

Hanau (1897), also working on fowls, found difficulty in completely cas-
trating cocks, and showed that a small amount of the testicle, if left behind,
forms little capsules, containing sperm and embedded in connective tissue.
Testicles transplanted into hens were encapsulated and resorbed.

In 1898 Goebell, questioning the results of Berthold, operated on g-uinea-
pigs and found that transplanted testes in the body-cavity became necrotic
after two days, except the most superficial layers, which showed mitoses.

Herlitzka (1899), in a very critical paper, reports the results of the trans-
plantation of testicles in tritons. In all, 32 animals were used, 20 females
and 12 males. These experiments were in part planned to test the theory
of Ribbert (1898), who, working on a large series of rabbits, found degener-
ation to take place, or more correctly a process of regression to a more
primitive type of cell, both when bits of tissue were placed in lymph nod-
ules and when whole testicles, autoplastic or homoplastic, were fixed by a
peritoneal suture. Ribbert found that the epithelium of the efferent canals
resisted longer than the epithelium with specific function. He concluded
that transplantation of the testicles was not possible because there is con-
cerned a gland which discharges its secretion externally.

To test this last theory Herlitzka used tritons both in winter stages and in
summer stages . The testicles were transferred from one animal to another,
and also to females ; and in some males one of the animal's own testicles
was left as a control. His results in brief were degeneration of the trans-
planted tissue under all conditions in from ten to fifty-two days, usually
about a month. The state of the testicle, whether resting or active, made

20 ON GERMINAL TRANSPLANTATION IN VERTEBRATES.

no difference. The efferent parts of the tubules degenerated, though more
slowly than the sperm-forming parts. The author is inclined to attribute
non-success, in part at least, to the absence of trophic stimulation of the
nervous system, though he admits his view is far from being proved.

Zalachas in 1907 gives the result of autoplastic transplantation in frogs,
in which he found living sperm preserved after one month. He does not
claim that this proves activity of the transplanted gland, for as a control
he found (completely degenerated but containing active spermatozoa) a
testicle which had been for one month in a sterile tube. Zalachas also
transplanted testicles of very young dogs into older dogs, and in seven
cases got almost complete degeneration at the end of one month.

Foges (1898) extended the result of Lode's work on caponizing cocks,
without obtaining any very convincing results.

Foa (1901), in a very able paper, sums up the results of others, and then
describes in detail his own work on dogs, autoplastic and homoplastic, which
gave negative grafts after one month. The organs of three-day old dogs
were used for the transfer. In another experiment a part of one of these
testicles was carefully sutured by the tissues to another testicle, in which,
though left in situ, a part of one end had been cut off. The cut faces of
the two testicles were thus brought face to face. Complete degeneration
took place both in the graft and in the remaining portion of the animal's
own testicle.

Foa therefore combats the theory of the lack of efferent ducts suggested
by Ribbert. His results are the same whether the testicle is embryonic or
adult, whether whole organs or parts of organs are used. Foa gives weight
to traumatism, brought about by the operator, as the essential cause of
degeneration.

We believe that the results of Maximow (1899), in the study of the regen-
erative capacity of the testicle after artificial wounds, show the testicular
tissue to be of a very delicate nature, easily unbalanced b> slight mechan-
ical disturbances. The work of Cevolotto (1909) goes far towards strength-
ening this view. This author made a careful study of the cell changes in
autoplastic transplantation of bits of testicular tissue into the tissue of the
ear. His results, in a word, show the harmful influence to the highly
differentiated epithelial tissue, which last is quickly converted into embry-
onic tissue. Sertoli cells increase and epithelial giant cells appear.

With this idea in mind the results of Guthrie (1910) are indeed surpris-
ing, though it must be confessed that a review of the literature shows the
cock to be a very likely subject for transplantation experiments. Guthrie
reports normal testicular structure existing in grafts after four months.
He hopes to produce artificial fertilization in hens from such testicular
grafts in continuation of his experiments on the effect of the foster-soma
on engrafted germ -cells.

GRAFTING OF TESTICLES. 21

7. NEW OBSERVATIONS ON GRAFTING OF TESTICLES.

In order thoroughly to satisfy ourselves whether g-erminal transplantation
is possible when tissue of the male sexual gland is used as a graft, the
following experiments were performed on rats before ovarian work was
started. The operations were homoplastic and the technique simple. The
animals used to supply the graft were new-born males or those only a few
days old, while those into which the tissue was grafted were young adults
of the same sex. No attempt was made to unite the efferent ducts, or to
supply special arterial contact, the young testicle being simply dropped
into the tunica vaginalis after its original contents had been carefully
removed.

The rat was etherized, the scrotum sterilized and opened at its discal
end by a small incision. The testicle was then drawn out, the tunic split,
and the entire contents removed, leaving- the end of the spermatic artery
free. The other testicle, now quickly removed from the young animal sup-
plying' the graft, was carefully manipulated and placed within this tunica,
sometimes with its own envelope intact and sometimes with one of its
ends cut across. The tunica, now containing the young grafted testicle,
was closed by fine silk sutures, the testicle replaced, and the scrotal wound
also closed. There was considerable extravasation of blood from the sper-
matic artery in a few cases, distending the tunic and sometimes leaking
through the sutures. The testicle of the other side was removed through
a small incision in the scrotum and discarded, a suture being placed around
its spermatic cord.

Thirty-three rats were thus operated upon. One died six days after the
operation; the rest were killed at periods of from one month to two months
after the operation. Four of them showed small abscesses in the operative
region. The other twenty-eight gave no evidences of ever having been
infected. In general, the appearance was very uniform. The epididymis
was flabby, and in the region of the graft there was either a very small
yellowish body or nothing but a bit of connective tissue surrounding the
old stitches of the tunica. A number of the specimens were saved, but
only one was considered to be promising enough to warrant sectioning.
The microscopic examination was entirely negative so far as showing the
presence of living testicular tissue was concerned, and so the experiments
were discontinued.

It is evident from our experience, as from that of others, that testicular
tissue is much more sensitive to injury, mechanical or otherwise, than is
ovarian tissue.

22 ON GERMINAL TRANSPLANTATION IN VERTEBRATES.

7. BIBLIOGRAPHY OF OVARIAN GRAFTING.
Arendt, E.

1898. Ovarientransplantationen. Verhandl. d. Geselisch. deutsch. Naturf. u. Aerzte,

Leipz., v. 70, pt. 2: 173. Also, Centralbl. f. Gynak., Leipz., v. 22 : 1116.
1906. Ueber Ovarialtransplantation (mit Demonstration). Verhandl. d. Geselisch.

deutsch. Naturforsch. u. Aerzte, Leipz., v. 77, pt. 2: 231.
Basso, G. L. 1905. Ueber Ovarientransplantation. Arch. f. Gynak., Berl., v. 77: 51-62.
Btjcura, C. J. 1907. Ueber Nerven in der Nabelschnur und in der Plazenta. Zeit. fiir

Heilkunde, Wien und Leipz., v. 28, Abt. fiir Chirurgie: 12-28.

BURCKHARD, G.

1908. Ueber Ovarientransplantation. Berl. klin. Woch., v. 45: 1337.

1908. Ein Beitrag zur Ovarientransplantation. (Transplantation von Ovarien indie

Hoden bei Kaninchen). Beitr. z. path. Anat., Jena, v. 43 : 499-518.
Also, Centralbl. f. Gynak., Leipz., 1908, v. 32: 150.
Carmichael, E. S. 1907. The possibilities of ovarian grafting in the human subject

as judged from the experimental standpoint. Jour. obst. and gynasc. Brit.

Empire, Lond., v. 11: 215-223.
Castle, and W. E. Phillips, John C. 1909. A successful ovarian transplantation in

the guinea-pig, and its bearing on problems of genetics. Science, n.s., v. 30:

312-314.
Chrobak, R. 1896. Ueber Einverleibung von Eierstocksgewebe. Centralbl. f. Gynak.,

Leipz., v. 20 : 521-524.
Croom, J. H. 1905-03. A case of heteroplastic ovarian grafting, followed by pregnancy

and a living child. Query: Who is the mother? Trans. Edinb. obst. soc,

v. 31 : 194-200.
Dauber, J. H. 1905. The ovarian ligament and its relation to pregnancy occurring

after ablation of the ovaries. Lancet, Lond., 1905 I (v. 168): 224-225.
Davenport, C. B. 1910. Inheritance of plumage color in poultry. Proc. Soc. Exp.

Biol. Med., v. vn: 178.
Doran, M. A. 1902. Pregnancy after removal of both ovaries for cystic tumor. Amer.

jour, of obstetrics, v. 46: 111-115.
Dudley, A. B.

1899. Intrauterine implantation of the ovary. Comptes-rendus du Cong. peYiod. in-

ternat. de gyndc. etd'obst., 1899 (Amsterdam, 1900), v. 3: 70-73.

1900. ^/^.Post-graduate, N. Y., v. 15: 546-548.

A further report on ovarian implantation. Cong, internat. d. me'd., Comptes-
rendus, Paris, 1900; Sect, de gyne'c: 387-388.

1901. Also, Jour. Amer. med. assoc, Chicago, v. 27: 357-360.

Fish, E. F. 1898-99. The uterus again. Annals of gynec. and pediat., Boston, v. 12:

379-385.
Foa, C.

1900. La greffe des ovaires, en relation avec quelques questions de biologie gdneVale.

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Die Schwangerschaft bei der Transplantation der Eierstocke. Centralbl. f.
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BIBLIOGRAPHY. 23

Guthrie, C. C.

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24 ON GERMINAL TRANSPLANTATION IN VERTEBRATES.

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8. BIBLIOGRAPHY OF GRAFTING OF TESTICLES.

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26 ON GERMINAL TRANSPLANTATION IN VERTEBRATES.

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