PIt E C 0 CIO U S S XU AL D E VELO 1' SlE N T F R O X

I N A MONKEY ANTERIOR HYPOPHYSTS IBIPLSNTS

O N E IIELIOTYPR r u m (FIVE FIGURES)

The recent demoiistrations by Smith ('26 b ) , Smith and Phigle ( '27) ' and Zoiideli aiid dscliheim ('27) of the depend- c1iice of ovarian function upon the secretion of the anterior lobe of the hypophysis has been especially convincing. Smith's ( '26 a ) success in inducing the recovery of atrophied gonads after ablation of the anterior hypophysis was fol- l o m d by an almost simultancous demonstration by Smith aiicl Engle and by Zoiidek and Aschheim that several implants of anterior lobes in immature mice greatly stimulated the de- rclopment of follicles in the ovaries, and that such implants precociously induced sexual maturity. By this treatment a superorulation was ohtainetl. Recently, Engle ( '27 b) has showii, by successful mating experiments in which consider- able numbers of o r a mere fertilized, that a large proportion of these ova are normal.

In the course of attempts to obtain experimental premen- strual and menstrual uteri in ovariectomized adult monkeys, MacacLls rhesus (Allen, '28 b), it became necessary to sacrifice several animals at two-day intervals. This made possible the extension of the hypophyseal experiments to primates. The monkeys from which hypophj-ses were taken mere killed by bleeding. As sooii as possible after death, the hypophy-

'Assisted by a grant froni thc Committee f o r llrsearcli on Problems of Sex of thc National Research Council and by D. D. Robertson, 1,. Bland, and I€. J,. I Ioorer , researcli assistants under this grant.

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316 EDGAR .4LLEK

sis was removed from the sella turcica and the posterior lobc dissected out. T l ~ e anterior lobe was then cut into small dices and implanted into small subcutaneous pockets in the flank of an immature monkey. A total of four implants was made a t two-day intervals. Three monkeys from which the anterior lobcs were removed had previously been ovariecto- mizcd arid then, following intervals of castrate atroph;v, had received iiijectioiis of‘ more than 550 r a t units of ovarian hor- mone during twenty-four days. They were killed on the first, third, and fifth days after the last injection. The last lobe was obtained from a very large seven-year-old male moiikcy from which one testicle had been removed several months prerions t o death. Smith and Engle have show~ii that the sex of the donor makes no difference in gonad-s timulation effects of anterior lobe.

Rcforc the first implant the immature host was laparoto- mizcd and ohservations made upon the size and vascularity of the uterus and the ovaries. The uterus mas very small (9 x 5.5 mm.) and nnemic. The ovaries were small and whitc and showed no signs of large follicles. Several medium- sized follicles could bc seen a t the snrface of each ovary. There were n o corpora lutea visible in either of the ovaries: coiisequcntly, ovulation had not occurred. This conditioii was to he expected, as the body weight of this monkey (3465 grams) was considerably less than the average body weight a t which monkeys in our colony have attained sexual maturity. In addition, there had been no developnierit of the secondary sex characteristics of reddening and swelling of the ‘ sexual skin.’ At this same operation a left mammary gland was rcmoved for a control and a control smear was taken of the vaginal contents.

The first implant was made on the 16th of February; the last, on the 22nd. By the 23rd, or the seventh day after +he beginning of the experiment, the ‘sexual skin’ was very red and considerably swollen. The, reddened, swollen condition extended over an area of approximately 7 X 6 em., and the skin and sabcntaneous tissue was 2.5 em. thick. This con-

SEXUAL DEVELOPMENT IN A MONKEY 317

Controls, immature Implanted, anterior

hypophyms

dition had developed more rapidly than is usual in the case in normal animals and more rapidly than is usual during injections of ovarian hormone into spayed animals.

Vaginal smears showed many nucleated epithelial cells, but very few or no cornified cells. These findings will be further discussed after a histologic description of the vaginal wall.

Before death, the body weight was 3504 grams-a gain of 39 grams in seven days. The uterus measured 15 X 11.3 mm. -an increase of 6 X 5.5 mm. in seven days. The uterine tubes were much more prominent and the ovaries were very much enlarged-surely, more than four or five times their size seven days previously. The weight of both ovaries was 1.418 grams as compared with 0.224 gram, the weight of the two

0.224 100 7.5X5.1X3.8 (3) 5.46 1.418 633 12.85X11.40X6.55 (2) 10.26

TABLE 1

Oaaries, weight and siae

ovaries of another immature monkey recorded in an earlier experiment. A comparison of size and weight is made in table 1. Many large follicles were apparent through the surface of the ovaries, but there were no signs of rupture points and no corpora lutea. A small piece was snipped from the fimbria of one of the tubes and observed in Ringer's solution under the binocular. Motile cilia were easily seen. The implants were dissected out and sectioned, the hypoph- ysis removed, and the right mammary gland dissected out for study.

The walls of the vagina were much thicker than those of the normal immature or spayed adult animals, but the outer zone was not cornified. As compared with experimental conditions following series of injections of ovarian hormone, vaginal

growth liatl not progi-essecl very far. Perhaps a longer iiiter- va l tliaii seveii clays is necessary for full growth of this tissue.

( ’erviciil glands w r e well developed, biit large numbers of Icuc*ocytcs were in the course of migrating toward the lumeii of tfirl uterus. This same condition has been foiuid in other aiiimals after stimulation with ovarian hormone. I t seems to the. writer that 110 rmone stirnillation must be considered its oiic of the mimy possible causes of leucorrhea.

IIistologic sections of the uterus sho~red the glands ell cieveloped and the connective tissue loose mvshed and c1tlcmatons. The enclornctrium had certainly developed consid- e>ixl)ly beyond the immature condition and might he charac- tcrizctl as approaching the interval stage of the adult.

flistologic section of the tnbes indicated considerable secrc- tory activity of the cpithelinm.

The ovaries a s studied in section had been transformed (figs. 4 ancl 5). T h ~ y contained maiij- large, medium, arid sm:~lI-siz~d follicles in which coiisiderable amounts of liqiior folliciili had heen elaborated. ‘L’hesc follicles were grouped i*oug%ly according to size. The large size can be seen at the 1owt.r pok of fignre 4 a i d in the npper. center of figme 3. Several mediumsized follicles are grouped at the upper polr of tlie sectioiis showii in figures 4 and 5. There were tv-enty- siy and thirty-one large and twenty-four and twenty-nine me- clinm follicles in the riglit and lcft ovaries, respectively, :ind morel than thirty small follicles in each ovary. 7’he contrast I)vtwcieii these ovarios and the ovaries of two other immature aiiimals of slightly less hoclp weiglif is sliown by comparisoii of figures 1 and 5 with figures 1 and 2. Figures 1 and 2 are fi-om two normal, immature monkeys and represent the con- t t.ol coidition (Allm, ’28). They coiitaiii the only follicles tlesigiiated in this tlcscsriptioii as small. The stage of devclop r r w n t of t l i ~ oi-aries after these implantt; of anterior. lobe is

ntnated by tlie contrast hetwmi them and a11

ov;\r\- from R maturc, animal s1io.is.n in fignrc 3 which con- t a i l i d two corpora lutcw, oiie recent one still showing the i ~ i 1 1 ) t i i i ~ i point, atid ;I sccond of loss thaii i hirt7 days’ tlewlop-

S E X V A L 1)EPELOPMENT I N A MONKEY 319

ment (Allen, '27). All of these five ovaries were sectioned in planes through the longest diameters.

The right (experimental) mammary gland had grown to more than twice the area of the left gland (control). The ducts were larger, there was an iiicreased number of alveoli, and the experimental gland, mounted in a flat spread, covered more than twice the area of the control.

Sections of the implanted tissue showed the first implant to have been almost completely disorganized. I t was difficult to recognize any hypophyseal tissue. The last two implants which had been in the host for one and three days, rcspec- tively, still contained considerable areas of normal hypo- pliyseal tissue, but they were for the most par t very head:- infiltrated with leucocytes.

After noting the size of the follicles and the thin layers of ovarian tissue separating a few of the largest from the surface of tlie ovary, it seems probable that one or two morc implants might have induced a superovulation in this animal.

In attempts to follow out this lead, two other immatim monkeys were subjected to series of daily implants of dog liypophyses. The experiment was carried along for eight days, and then the animals were opened to observe possible effects. 1'0 o w surprise, the ovaries were small and undc- veloped and the uteri infantile. Implants from the dogs liacl a serious effect upon the licalth of these monkeys, both were very ill aiid one died several days after the last implant.

The apparent noii-effect of the anterior lohe from the dogs is interesting. From much previous worli the expectation would be that this hormone ~vould be active between different species (Englc, '27 a ) . I-larmful effects of foreign tissue protein of tlie dog may liave been a contrihuting C R L W in failure of these experiments. The questioii arises, however, a s to whether tlie anterior lobe in the dog may not be less active than that of the monkey, as f a r as this gonad-stimulat- ing substance is concerned. It may be that such an explana- tion wonld accoimt for the rarity of recurrence of oestrns and the long diocstrous interval ill most dogs.

320 EDGAR -4LLEN

SUMMARY AND CONCLUSIONS

A series of four implants of anterior lobes of hypophysis T\TC’IT made at two-day intervals in an immature female mon- key. In seven days this treatment caused marked develop- ment of secondary sex characteristics and considerable growth in the genital tract and mammary glands. The ovaries were greatly enlarged, due to a marked stimulation of growth of follicles with format ion of liquor folliculi. About thirty larye follicles had developed in each ovary. Some were apparently very near t o ovulation.

The effects in sex characteristics and growth of the genital tract and mammary glands are similar to those obtained in normal and castrated monkeys from injections of ovarian and placental hormone. These effects were apparently sec- ondary to the marked ovarian development, for, as noted by Smith and Engle, anterior-lobe implants have no effect upon castrated animals. Similar experiments with implants of anterior lobe from dogs gave negative results. It is concludcd that the anterior lobe of the monkey contains considerable gonad-stimulating hormone, while that of the dog seems to contain li ttlc.

LITERATI’RE CITE11

, E. 1927 The mcnstrual cycle of the monkey, Macacus rhrsus: observa tions on normal animals, the effects of removal of the oiaries and the cffrrts of injections of ovarian and placental rxtracts into the spayed aniinols. Contrih. t o J3mbryol., Carnegic Institution o f Washington, \ol. 19, no. 380, pp. 1-44.

Reactions of the inimature nionlrcy, Macarus rhesus, to in- jcctions of an ovarian hormone. Jour. hlorph. and Phjsiol. ( I n press.) - 1928 h Further experiments with an ovarian hormone in the oiari-

ectoniized adult monkey, Mscacus rhesus ; especially the degeneratirc phase of the expeiimental nicnstrual cycle. Am. Jour. Anat. ( in press)

I’:ZQLE. B. T. 1927 a Gonad-stimulating hormone of anterior pituitary and heterosexual ovarian grafts. Procs. Soc. Exper. Rial. and Med., vol. 25, p. 83.

__-__ 1927 h Prrgnanry following super-ovulation in the mouse. Procs. Soc. Exper. Biol. and Med., 101. 2.5, p. 85.

S w n r , P. E. 1926 a Ablation and transplantation of the hFpophysis in the rat.

~ 1928 a

4na t . Rw., vol. 3 2 , p. 227.

SEXUAL DEVELOPMENT IN A MONKEY 32 1

SAIITH, P. E. 1926 b Hastening dcvelupment of female genital system by daily hornoplastic pituitary transplants. Prom. Sor . Exper. Biol. and Med., vol. 24, pp. 131, 132.

SMITH, P. E., AXD E. T. EPTGLE 1927 Experimental evidcnce regarding the rfile of the anterior pituitary in the development and regulation of the genital system.

Das Hormone des hvpophgsenvorderlap- pens.

Klin. Wochenschr., Bd. 6, S. 1321- 1322.

Am. Jour . Anat., vol. 40, pp. 159-217. ZONDEIC, B., AND S. ASCHIIEIN 1937 a

Klin. Woehensehr., Bd. 6, S. 248-252. 1927 b Ovum and hormones.

PLATE 1

lCXPLAh’ATION OF FIQUKES

Sections of all ovaries were made through t.he longest diamc.t.ers. Figures 1 and 2 arc from ovaries o f two normal imrnaturc animals of anothrr

series which served a8 controls. Figure 3 is from a large ovary of a iiomal adult monkey containing two

c*orporr, one of which, being ver7 rcccat, still shows a rupturo point, nherers the ot.hcr is probably less than thirty days of age.

Figures 4 and .5 are from thc two ovaries of the inimature animal which rc.c.c.ivecl four Imterior-hSpol)liysie intplants.

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REX1J.U. DEVLLOPMENT I N A MOhTKE\ EDQAH. ALLEN

323

PLATF. 1