Bull Testis Research Articles

STUDIES ON THE INHIBITORY HORMONE OF THE TESTES IV. EFFECT ON THE PITUITARY, THYROID AND ADRENAL GLANDS OF THE ADULT MALE RAT1

THE PREPARATION of an extract, of bull testes, which produces inhibitory effects on the gross and microscopic appearance of the sex organs of adult male rats has been described (1, 2). In this paper we will consider the effect of this extract on the weight and microscopic appearance of the pituitary, thyroid and adrenal glands of the adult male rat. The data reported herein were derived from the pituitary, thyroid and adrenal glands of the series of rats we reported on previously (1, 2). The weights of the glands and the bodyorgan values were obtained in the same manner as reported in the former papers except that each bodyorgan value was divided by 100 in order to facilitate the reading of the tables. The tissues were fixed in formalin Zenker solution and embedded in paraffin. The thyroid and adrenal sections were cut at 10 micra and stained with hematoxyiin-eosin.

THE RECOVERY OF TESTES AFTER ANDROGEN INDUCED INHIBITION1

IN 1932, MOORE AND PRICE (1) showed that the administration of an androgenic extract of bull testes to immature rats had an inhibitory effect on the testes. The same effect wasobtained by these workers with crystalline androsterone (2), testosterone (3) and testosterone propionate (3). The “grave retardation and injury to the testes of young normal males” thus obtained consisted of gross decrease in size of the testes, reduction in the size ofthe tubules, sloughing of cells into the lumen and delay in the development of complete spermatogenesis. Similar findings were reported by Sainton, Simonnet and Petresco (4). While no statement was made as to the after effects of this treatment on the testes, the casual inference was that the damage would be permanent. It seemed pertinent to repeat this work in part and to observe the possible effect of this early treatment on later adult testicular function.

STUDIES ON THE INHIBITORY HORMONE OF THE TESTES1III. HISTOLOGICAL EFFECTS ON THE MALE SEX ORGANS OF THE WHITE RAT

THE PREPARATION of an extract from bull testes containing inhibitory properties has been reported in a previous publication (1). The effect of this extract on the weight of the male sex organs has been described (2).The present paper will be limited to a study of the effect on the histologic structure of the testicle, seminal vesicle and prostate of animals injected with this extract. The glands used for this study were those whose weight changes were previously reported (a). The normal histology of the rat prostate and seminal vesicle has been adequately described by C. R. Moore and his coworkers (3, 4). The division of the prostate referred to as the middle lobe by these authors is described by us as the lateral lobe. Since the lateral and posterior lobes are histologically similar, only the lateral lobe has been included in this report. The histology of the anterior lobe is not reported

THE URINARY EXCRETION OF ESTROGENIC SUBSTANCES AFTER THE ADMINISTRATION OF TESTOSTERONE PROPIONATE1

TRANSFORMATION of testosterone, the male sex hormone isolated from bull testis tissue, into a substance possessing estrogenic activity, has been claimed by Steinach and Kun (1) to occur in men. No such transforma-tion was found by Kochakian (2) in the dog, or by Dorfman (3) in the rat. Steinach and Kun were able to increase the urinary excretion of estrogenic material in the urine of men after the injection of testosterone propionate. In one case they report an increase in estrogenic activity from approximately 36 to 1200 R.U. per liter after the injection of 1 gm. of testosterone propionate during a period of 6 weeks. Their assays for estrogenic activity were made on the crude benzene extracts so that it is difficult to say what the nature of the excreted estrogenic substance may be. Further, the possibility exists that this activity may be due to androgens such as androstenediol or de hydroisoandrosterone, which are also capable of producing estrogenic effects.

THE USE OF TESTOSTERONE PROPIONATE IN FUNCTIONAL BLEEDING

The present communication is a report on the use of testosterone propionate (male sex hormone) in functional uterine bleeding. Of 25 cases so treated, the excessive bleeding was rapidly controlled in all but two. This clinical improvement was correlated with definite changes in the endometrium, as revealed by suction biopsies performed at intervals before, during and after treatment. The rationale for this form of therapy has its basis in animal experimentation as well as in our studies on the effect of testosterone propionate on the endometrial cycle of the human. Suppression of the estrus cycle in female rats was first induced with male hormone extracts of bull testes by Ihrke and D'Amour (1). Similar results were obtained with testosterone by Robson (2) in mice and Browman (3) in rats. In monkeys, Hartman (4) has shown that menstruation was delayed following testosterone administration, and Zuckerman (5) has reported that testosterone propionate inhibits follicle stimulation and luteinization, resulting in suppression of the menses. Cotte, Martin and Mankiewicz (6) reported similar results in rabbits. In a previous report (7) we have shown that administration of adequate doses of testosterone propionate causes striking morphologic changes in the endometrium of regularly menstruating women, associated with suppression of the succeeding menstrual period. These changes consist of the disappearance of the secretory phase and inhibition of the proliferative phenomena. The inhibition of proliferative activity in the endometrium, together with the suppression of the menses, suggested the use of male hormone in functional uterine bleeding. The hormone selected for this investigation was synthetic testosterone propionateinasmuch as it has been shown to be the most potent preparation available (8, 9).

THE EFFECT OF TESTOSTERONE PROPIONATE ON THE GENITALIA, PROSTATE, SECONDARY SEX CHARACTERS, AND BODY WEIGHT IN EUNUCHOIDISM1

With the synthesis of testosterone from cholesterol by Butenandt and Hanisch (1), and by Ruzicka and Wettstein (2), and the successful largescale commercial application of their processes, it is at last possible to undertake substitution for the testis in man. Although not as yet established as the actual hormone, testosterone has been obtained from bull testis by David, Dingemanse, Freud and Laqueur (3) and, with the exception of certain of its own esters, is the most potent of known androgens. It will repair castration defects practically completely in male animals. The literature dealing with this evidence has been reviewed recently by Koch (4) and need not be dealt with here. In 1936, Miescher, Wettstein and Tschopp (5) discovered that various fatty acid esters of testosterone not uncommonly stimulated the prostate and seminal vesicles of the rat more than did testosterone itself. This was true especially of the formate, acetate, propionate and butyrate, and not at all of the palmitate and stearate wh...

Effect of Testosterone Propionate upon Endometrial Cycle of the Human

Experimental evidence is accumulating which indicates that male hormone counteracts the physiologic effects of estrogenic hormone. Ihrke and D'Amour1 reported suppression of the estrous cycle in female rats while being injected with male hormone concentrates prepared from bull testes. Similar results were obtained by Robson2 in mice and Browman3 in rats with testosterone. Zuckerman4 has reported that testosterone propionate inhibits follicle growth and luteinization in monkeys. The present investigation was undertaken to determine the effect of testosterone propionate upon the endometrial cycle of the menstruating human female.A group of 6 women were selected and preliminary control endometrial (suction) biopsies were performed in each case to determine the presence of a normal endometrial cycle. In all of the cases selected for this study, the control biopsies performed during the latter half of the cycle showed, on one or more occasions, an endometrium in the secretory phase, indicating a regular estrog...

INHIBITION OF GONADOTROPIC ACTIVITY BY SEX HORMONES IN PARABIOTIC RATS

There seems to be no doubt that castration in the rat results in an increase in the gonad-stimulating capacity of the hypophysis, and it is also generally accepted that estrogenic substances depress the gonadotropic power of the pituitary (1). Moore and Price have shown that bull testis extracts (2) and androsterone (3) injected into young male rats cause testicular damage. They attributed this injurious effect to a lowering of pituitary gonad-stimulating hormone, an idea supported by the fact that implantation studies showed the pituitaries of androsterone-injected rats to be reduced in gonadotropic potency. Of special interest are recent reports (4, 5) that ovarian weights in immature parabiotic rats could be used as a quantitative measure of hypophyseal gonadotropic secretion of castrated parabionts which were untreated or treated with estrone or androgenic substances. Our own observation (6, 7) on paraboitic rats suggested the possibility that the testicular response of hypophysectomized rats might se...

A Pressure Device for the Separation of Mammalian Spermatozoa from the Isolated Epididymis

During an investigation of the specificity of mammalian spermatozoa it was desired to obtain a pure suspension of spermatozoa for purposes of chemical fractionation. As a very large quantity of material was needed, it was necessary to devise a means of separating them from the isolated epididymis with a minimum of labor. The following method, using a pressure device, has been found useful in rapidly separating the spermatozoa from a large number of the organs. The pressure device consists of a foot-pedal which activates a pressure bulb fitted with 2 one-way valves in line, a water reservoir, and pressure tubing leading to a blunt No. 22 gauge hypodermic needle fitted with a shut-off valve. The testes of bulls and rams were obtained from the abbatoir with all their coverings intact. After washing off all adherent blood clots, the cavity of the tunica vaginalis was opened and the cauda epididymis removed together with a long section of the vas deferens. These pieces of epididymis with the adherent vas deferens were placed in sterile saline until used. The open end of the vas deferens was pulled over the blunt needle, and secured by means of a clamp, the jaws of which were covered with fine rubber tubing, to prevent cutting of the tissues and back-flow. Pressure was then applied by means of the foot-pedal and water was injected into the epididymis. After injecting a small amount of water, the canal protruded in certain fields, one field appearing after the other over the shiny surface of the epididymis.

BIOCHEMICAL STUDIES ON THE MALE HORMONE AS OBTAINED FROM URINE*†

The testicular hormone obtained from human male urine has been the subject of investigation by many workers, notably Funk, Harrow, and Lejwa (1, 2, 3), McCullagh, McCullagh, and Hicken (4), Dodds, Greenwood, and Gallimore (5, 6) and Butenandt (7). With the exception of the last mentioned author, the extracts described by these workers have been rather crude compared to the relatively high purity of the bull testis extracts described by the present authors. The purpose of this communication is to present a method for the extraction and purification of the comb-growth-stimulating substance found in human male urine and to compare the chemical and physiological properties of this substance with the extracts from bull testis. The urine is acidified with sulfuric acid within two hours after collection to approximately 1 per cent by volume. After filtering through cloth to remove foreign particles, the urine is extracted with benzene in a continuous extractor as shown in Figure 1. When a sufficient quantity has...

SPONTANEOUS ACTIVITY IN MALE RATS IN RELATION TO TESTIS HORMONE*

The preparation by McGee (6) of extracts of bull testes that stimulated growth of the comb of castrated cocks and the subsequent advances in the purification of these extracts by Gallagher and Koch (1, 2) are now well known. Similarly, through the publications of Moore and his associates (7, 8, 9), it has been demonstrated that these extracts prevent the onset of castration changes in the mammal, or repair them if administered subsequent to castration damage [Vatno (14), Heller (4)]. Several mammalian test methods have been utilized, all of which have dealt with the accessory organs of reproduction. Since it is necessary to know that these preparations are effective in preventing all changes known to follow castration before arriving at the conclusion that they contain the complete hormone, I have made a study of the spontaneous activity in the rat in relation to its hormone state to learn whether voluntary activity can be successfully used as a hormone indicator.

Significance of Female Sex Hormone Reaction in the Male Blood.

In 1925, simultaneously and independently, Loewe of Dorpat1 and one of us with collaborators2 demonstrated the presence of the female hormone in the circulating blood of females by means of the rodent vaginal spread test.3 Since then, in numerous publications we have attempted to simplify and standardize the method of extracting and testing human blood for the female sex hormone.4 Among other applications we advocated the use of this test to determine the sex of pseudo-hermaphroditic individuals in whom we regarded a positive reaction, appearing cyclically, as a proof of the presence of functioning ovaries and feminine sex.5 Our preliminary work had shown that large quantities of bull's blood (150-100 cc.) gave a negative reaction when extracted by our method. The same applied to concentrated lipoid, HCl, saline and watery extracts of bull's testes, as well as extracts of the hypophysis, thyroid and adrenal, liver, muscle, various proteins, etc.6 The work of Dohrn,7 who claimed to have obtained a positive...