Morphological alterations and intratubular lipid inclusions as indicative of spermatogenic damage in cimetidine-treated rats

Doses of cimetidine (50 mg/kg b/w) were administered to adult male Wistar rats over 52 consecutive days. Besides plasma testosterone levels, morphological and morphometric aspects of the seminiferous tubules as well as histochemical analysis of the lipid content by oil red O were emphasized. Abnormal tubules exhibiting disorganization of their cellular association, loss of germ cells, and multinucleated giant spermatids were usually found. Significant reductions of testis weight and tubular diameter at specific stages (VII-IX), as well as lack of contact between Sertoli cells and spermatids in tubules at stage IX, suggest a possible interference of cimetidine on the histoarchitecture of the seminiferous epithelium. The dense concentration of lipid inclusions in tubules at postspermiation stages indicates phagocytosis and degradation of germ cells. Since no change in serum testosterone levels was verified in cimetidine-treated rats, the authors could not exclude the possibility that besides an antiandrogenic effect, other biochemical factors necessary for normal spermatogenesis could be involved in the testicular alterations.     

Effects of gonadotrophin treatment in vivo on testicular function in immature rhesus monkeys (Macaca mulatta)

Changes in testicular histology and concentrations of testosterone and oestradiol 17β in testicular tissue and plasma have been studied following administration of gonadotrophins (oFSH, oLH, hCG and PMSG) to immature male monkeys. Treatment with FSH (1 mg/day) or PMSG (100 IU/day) for five days, induced a marked enlargement of the seminiferous tubules and increase in the Sertoli cell cytoplasm. Injections of LH (1 mg/daily) or hCG (100 IU/daily) administered similarly, failed to produce hypertrophy of the Sertoli cell. In LH, hCG and PMSG stimulated testes morphologically differentiated interstitial cells could be recognized. FSH did not produce any detectable effect on the intertubular tissue. A significant increase in testicular and plasma testosterone levels was observed with LH, hCG and PMSG. FSH was shown to be much less effective in stimulating androgenesis. An increase in testicular oestradiol production over that of controls, was observed in FSH and PMSG treated monkeys but not in animals treated with LH or hCG.     

Local regulation of Leydig cells by the seminiferous tubules. Effect of short-term cryptorchidism

Unilateral cryptorchism was induced in adult rats for 24 h, and its effect on testicular morphology and intratesticular testosterone concentration after hCG-stimulation were studied. In seminiferous, tubules from abdominal testes an increased number of degenerating germ cells was noted in stages XIV-III of the spermatogenic cycle and Sertoli cells contained an increased amount of lipid droplets in stages XIV-VIII. However, germ cells and Sertoli cells from tubules at other stages of the cycle appeared unaffected. In scrotal testes the size of peritubular Leydig cells varied in phase with the spermatogenic cycle. The largest cells were found adjacent to stage VII-VIII and the smallest adjacent to stage XI-XII. In abdominal testes no stage-dependent variation in the size of peritubular Leydig cells was seen. Perivascular Leydig cells were of equal size in abdominal and scrotal testes. The testicular testosterone concentration following stimulation with a low dose of hCG was significantly lower in abdominal testes. It is suggested that the seminiferous tubules locally modulate Leydig cell function and that the stage specific stimulatory influence from stage VII-VIII is rapidly lost during experimental cryptorchidism.     

On the Postnatal Development of the Terminal Segment of the Seminiferous Tubules in Normal and Germ Cell Depleted Rat Testes

The development of the terminal segment of the seminiferous tubules was studied in 5 to 50 days old normal rats. At the age of 5, 10, and 15 days the terminal segment contained fewer gonocytes or spermatogonia than did the corresponding seminiferous tubule. The differentiation of the terminal segment was obvious at 20 days of age due to the high number of germ cells in the seminiferous tubules, where the epithelium became stratified at this stage. The blood-testis barrier in the terminal segment was chiefly established between 15 and 20 days of age as revealed by the lanthanum tracer technique.
To study the effect of the germ cells on the differentiation, the germ cell depleted testes of prenatally irradiated rats were also studied. The modified Sertoli cells of the terminal segment were more vacoulated and had fewer lipid droplets and inter-Sertoli cell junctions than did the Sertoli cells of the seminiferous tubules. The ultrastructure of the modified Sertoli cells of the terminal segment was similar in adult normal and adult SCO (Sertoli cell only) rats. The amount of lipid droplets in the Sertoli cells of SCO rats showed considerable variation among different tubular cross-sections within one testis.     

In-vitro contractility of human seminiferous tubules in response to testosterone,dihydrotestosterone and estradiol

Summary The effects of steroids (testosterone, dihydrotestosterone and estradiol) on human seminiferous tubules in vitro were ascertained by recording the intratubular pressure with a servonull micropressure measuring device. We describe here the first response of the human seminiferous tubule to steroids. Testosterone and dihydrotestosterone had a biphasic effect on tubular contractility. Higher doses of both testosterone and dihydrotestosterone induced contractions of the seminiferous tubules whereas lower doses of these compounds induced relaxation. Estradiol (10^-9 M to 10^-6 M) induced relaxation of the seminiferous tubules in a dose-dependent manner. The results from these experiments suggested that steroids may be involved in the control of contraction of the human seminiferous tubule and may regulate the movement of spermatozoa from the tests.     

Evolution of somatic and germ cell populations after busulfan treatment in utero or neonatal cryptorchidism in the rat

Summary.  In order to elucidate the respective effects of depletion of germ cells and of increase in testicular temperature, rats of the same Wistar strain were rendered experimentally bicryptorchid or sterilized by a busulfan injection in utero and compared to control animals. In both models, germ cells were depleted but numeric evolution and functions of somatic cells differed. The aim of that work was to compare the numeric evolutions of testicular somatic and germ cells to their respective functions in each model before puberty and in adult rats of the same strain. Serum concentrations of FSH, LH and testosterone were compared at 20, 40 and 110 days of age. Histological analyses of Sertoli and germ cells in the seminiferous tubules and of Leydig cells in the intertubular tissue were performed before puberty and at adulthood. Testosterone serum concentrations were depleted in both models starting at 40 days of age and more in busulfan-treated rats. Both FSH and LH levels were increased from 20 days onwards in experimental rats. Additional cryptorchidism in busulfan-treated rats depressed the serum testosterone concentration. At 17 days of age, the cryptorchidism do not modify somatic or germ cell populations while busulfan treatment has induced a decrease of both these populations. Conversely, the cross sectional area of the somatic testicular cells was not affected whatever the treatment. In adult testes of busulfan-treated and cryptorchid rats, the total numbers of Sertoli and Leydig cells and of germ cells per testis were decreased. The cellular size of the perivascular Leydig cells was not modified by any of the treatments whereas the size of the Sertoli cells was reduced.
In conclusion, in both models the absence of germ cells induces a decrease in Sertoli cell function, while the increase in testicular temperature provokes degeneracies of Sertoli and germ cells in the seminiferous tubules of the rat.     

Steroidogenesis by Granulosa and Sertoli Cells

Granulosa and Sertoli cells have been isolated from the gonads of immature rats, and have been maintained in monolayer cultures in a chemically-defined medium. The hormonal requirements of these cell types for the synthesis of steroids have been studied: Granulosa cells contain cholesterol side-chain cleavage activity and synthesise progesterone when cultured in the presence of FSH and testosterone (or DHT). Sertoli cells on the other hand cannot synthesise steroids de novo . Granulosa and Sertoli cells are similar in that they are unable to convert significant amounts of progesterone to androgens. In the presence of exogenous testosterone as substrate, granulosa cells and immature Sertoli cells can synthesise estradiol and estrone when stimulated with FSH. In summary, FSH stimulates estrogen synthesis from testosterone in both cell types, and acts synergistically with testosterone (or DHT) to stimulate progesterone synthesis in granulosa cells. FSH is therefore involved in controlling the steroid environment within the gonads.     

Testicular Biopsy and Hormonal Study in a Male with Noonan''s Syndrome

The testicular biopsy study of a 17-year-old male with Noonan's syndrome revealed seminiferous tubules of reduced diameter with hypospermatogenesis. Many spermatocytes underwent degeneration and many spermatids developed abnormal. The Sertoli cells were similar to immature Sertoli cells. Fully differentiated Leydig cells were rare while precursor Leydig cells were numerous. Both gonadotropin and testosterone levels were low, and a lack of response to LH-RH as well as to clomiphene was found. The testicular biopsy performed at 20 years of age revealed a certain maturation of the seminiferous tubules which increased the germ cell number. The abnormalities in the spermatogenesis as well as the immature appearance of Sertoli cells continued. Leydig cells were more numerous and showed a certain development without reaching the normal pattern. Gonadotropin levels were normal while testosterone levels low. The response to LH-RH was increased and the absence of response to clomiphene persisted. These features suggest a delayed puberty.     

Ontogeny and cellular origin of a rat seminiferous tubule factor(s) that inhibits LH-dependent testosterone production by interstitial cells in vitro

Previous studies have shown the presence of a peptide in spent media from incubated seminiferous tubules (SMST), which inhibits LH stimulation of testosterone production by rat Leydig cells in vitro. The present study has investigated whether the secretion of this inhibitor changes during development in the rat. Seminiferous tubules obtained from rats aged 10, 20, 25, 30, 35, 40, 42, 50 or 60 days were incubated at 32 degrees C for 24 h. Spent media from these incubations were then added to interstitial cells isolated from the testes of rats aged 60 days. Spent media from rats aged 10-30 days had no effect on basal or oLH-stimulated testosterone production by interstitial cells during 3-h incubation. Significant inhibition of LH-stimulated testosterone production was, however, observed with SMST from rats aged 35-60 days. Spent media prepared using tubules from normal, prenatally irradiated (Sertoli cell-enriched) or seminiferous tubules, depleted of peritubular cells, had no effect on basal, but inhibited LH-stimulated, testosterone production. Spent media from peritubular cell cultures had no effect on basal or LH-stimulated testosterone production by interstitial cells. The inhibitory effect of SMST was also dependent on the age of the rats providing the target cells. Interstitial cells from rats aged 10, 20, 50 or 60 days were responsive to the inhibitor while cells from rats aged 30 and 40 days were not. The results of the present study demonstrate that the seminiferous tubule factor(s), which inhibits LH action on interstitial cells, is first secreted at 35 days, a time when the most mature germ cells present are in the early maturation phase. Moreover, interstitial cells are responsive to this factor in both immature (10-20 day-old) and mature (50-60 day-old) rats, but not at ages in between these times. It is suggested that the adult Sertoli cell is the major source of the interstitial cell inhibitor.     

Effects of diazacholesterol dihydrochloride (SC-12937), an avian antifertility agent, on rat testis

The present study was undertaken to evaluate the effectiveness of an avian chemosterilant, 20, 25-diazacholesterol dihydrochloride (SC-12937), on the rat testis. Adult male rats were injected intraperitoneally with 10 mg (Group 1) or 30 mg (Group 2) of SC-12937/kg/d or with vehicle alone (Group 3) for 10 days, and were killed 24 hours after the last injection. A wide range of variation in the appearance of affected seminiferous tubules was observed in the testis of SC-12937-treated rats at both dose levels. This ranged from apparently normal-looking seminiferous tubules to almost completely atrophied tubules with no cells. Affected tubules exhibited intraepithelial vacuoles of varying size, multinucleated giant cells, germ cell exfoliation, and tubular atrophy. The presence of severely damaged and entirely normal seminiferous tubules adjacent to one another in the same section was noteworthy. The changes appeared to be dose-related. A greater number (34.6%) of affected tubules were observed in rats receiving 30 mg of SC-12937 compared with the ones receiving 10 mg of this compound (19.6%). The Sertoli cells also were affected by this drug and exhibited cytoplasmic vacuolation, a marked increase in the accumulation of lipid droplets and myeloid bodies. Necrotic Sertoli cells also were observed in the severely affected tubules. The possible mechanism of antispermatogenic action of SC-12937 in rats has been discussed briefly.     

Spermatogenic onset. II. FSH modulates mitotic activity of germ and Sertoli cells in immature rats

By using high doses of testosterone propionate (TP) endogenous FSH was lowered to non-detectable levels in immature rats of different ages. Combined administration of TP and human menopausal gonadotrophin (hMG) or purified human FSH (hFSH) restored circulating FSH to normal or supranormal levels. This experimental model was used to investigate the influences of hormones on the proliferation of Sertoli and germ cells. Mitoses in seminiferous tubules were counted after being blocked by administration of colchicine. The mitotic index so determined showed a reduction with FSH withdrawal and a significant increase when the hormone levels were restored. Testicular DNA content was determined in testicular homogenates and showed similar changes. Autoradiographs were prepared and 3H-thymidine incorporation into germ and Sertoli cells was quantified. It was found that hFSH induced a significant increase in the labelling indices of gonocytes, type A-spermatogonia and Sertoli cells. All of these changes were effective in rats younger than 10 days but no modifications in any of the parameters under study were observed after 20 days. It is concluded that FSH exerts a stimulatory effect on the proliferation of spermatogonia, Sertoli cells and testicular DNA content during the first 10 days of life.     

Morphological and Histochemical Observations on the Prenatal and Postnatal Testes of the Field Rat (Millardia meltada)

The foetal testis of the field rat (Millardia meltada) shows seminiferous tubules and interstitium consisting of mesenchymal cells and differentiated Leydig cells associated with blood vessels. The tubules during the prenatal period show gonocytes and Sertoli cells. After birth, their diameter decreases but again increases progressively after postnatal days 9 and 10. Spermatogonia appear among the gonocytes on postnatal day 1 and primary spermatocytes on day 8. Secondary spermatocytes and spermatids are not seen up-to-day 23. Several hypertrophied Leydig cells are seen in the foetal testis on day 17 but are greatly increased in number on days 18 and 19. A few hours after birth the Leydig cells show a rapid decrease in their number. These fluctuations in the Leydig cells of prenatal and neonatal testes have been correlated to the rise and fall in testosterone production during these periods. The Leydig cells show the histochemical characteristics of actively steroid-secreting cells, which consist of the presence of diffuse lipoproteins and a few lipid granules consisting of phospholipids; no cholesterol and/or its esters could be demonstrated. Such lipids are not present in the cytoplasm of undifferentiated mesenchymal cells. The seminiferous tubules do not show any appreciable development of lipid changes.     

The Terminal Segment of the Seminiferous Tubules and the Blood-Testis Barrier Before and After Efferent Ductule Ligation in the Rat

The ultrastructure of modified Sertoli cells in the terminal segment of the seminiferous tubules in the rat was studied in material fixed by perfusion through the thoracic or the abdominal aorta. The central lumen of the terminal segment was not distinct under normal conditions, but became quite evident with an increased intratesticular pressure caused by efferent ductule ligation. The modified Sertoli cells were characterized by a marked increase in number of microtubules and microfilaments, extensive inter-Sertoli cell junctions, vacuolated cytoplasm, and considerably dilated intercellular spaces. Three types of specialized cell contacts were found between modified Sertoli cells: typical inter-Sertoli cell junctions, desmosome-like devices, and septate-like tight junctions. These specialized cell contacts were efficient in preventing lanthanum to penetrate into the lumen of the terminal segment both before and after efferent ductule ligation. Moreover, it was found that typical inter-Sertoli cell junctions prevented lanthanum penetration into the adluminal compartment of the seminiferous tubules even after efferent ductule ligation.     

Androgen effects on Sertoli cells

Mature male rats, gamma-irradiated in utero, were hypophysectomized. In an effort to maintain the seminiferous epithelium, some animals were treated with exogenous androgen while in other animals the seminiferous epithelium was allowed to regress without hormonal treatment. These Sertoli cell-enriched (SCE) males were evaluated for 7 weeks following hypophysectomy. In SCE males the average initial weight of each testis was 300 mg which declined to 110 mg at 7 weeks post-hypophysectomy. Concomitantly, seminiferous tubule diameter decreased from 130 microns to approximately 89 microns. Numerous cells were detached from the lamina propria and were observed in the centre of the tubule. Two layers of Sertoli cell nuclei were frequently observed in the regressed seminiferous tubules. Many of these nuclei appeared to be less differentiated, i.e. the nuclei were smaller with smaller nucleoli and more heterochromatin. In contrast, hypophysectomized animals treated with testosterone propionate during the last 5 weeks of the 7 week observational period, retained a tissue weight of about 270 mg/testis (a 5-10% decline in weight compared with normal untreated controls). Also, these animals had seminiferous tubule diameters of 132 microns. Finally, the Sertoli cells which comprised primarily a single layer inside the seminiferous tubules had larger nuclei with finely granulated chromatin and large nucleoli. Protein changes in SCE testes, (+/-) androgen, following hypophysectomy were analysed using polyacrylamide gels containing SDS. Prominent changes in the protein profile as separated by molecular weight were observed and were attributable to androgen stimulation. These changes were probably occurring in Sertoli cells since the Sertoli cell represents about 70% of the total cell population in the gamma-irradiated model. It is concluded that testosterone is responsible for major changes in mature Sertoli cells, although potential contributions of other cell types such as myoid cells and Leydig cells are considered.     

Suppression of pituitary and testicular function in rats by a transplanted adrenocortical carcinoma

Snell adrenocortical carcinoma was transplanted into immature 4-week-old male rats, and the animals were sacrificed 3 weeks afterward for study of adenohypophyseal and testicular function. The weight of the tumour was 11pL5 g; plasma corticosterone levels were elevated and plasma progesterone levels were massively increased compared to those in rats with no tumours. The weights of the adrenals, testes and androgen-dependent accessory reproductive glands were significantly decreased, as was the diameter of seminiferous tubules. The concentrations of testosterone, LH and FSH in the plasma were significantly reduced, whereas the levels of Prl and oestradiol were not affected. We suggest that steroid products of the transplanted tumour suppressed release of gonadotrophins from the pituitary, leading to a severe reduction of testosterone synthesis in the testes.     

Experimental diabetes negatively affects the spermatogonial stem cells’ self-renewal by suppressing GDNF network interactions

The present study was done to analyse the time-dependent effects of diabetes on Sertoli cells–spermatogonial stem cells’ (SSCs) network interaction by focusing on glial cell line-derived neurotrophic factor (GDNF) and its special receptors, gfrα1 and c-RET as well as the Bcl-6b. In total, 40 Wistar rats were considered in; control, 20, 45 and 60 days diabetes-induced groups. An experimental diabetes was induced by STZ. The GDNF, gfrα1, c-RET and Bcl-6b expressions were evaluated. The serum level of testosterone, tubular repopulation (RI) and spermiogenesis (SPI) indices, general histological alterations, germ cells, mRNA damage, sperm count and viability were assessed. The diabetes, in a time-dependent manner, diminished mRNA and protein levels of GDNF, gfrα1, c-RET and Bcl-6b versus control group (p < .05), enhanced percentage of seminiferous tubules with negative RI, SPI, and diminished Leydig and Sertoli cells distribution, serum levels of testosterone, sperm count and viability. Finally, the number, percentage of cells and seminiferous tubules with normal mRNA content were significantly (p < .05) diminished. In conclusion, as a new data, we showed that the diabetes by inducing severe mRNA damage and suppressing GDNF, gfrα1, c-RET and Bcl-6b expressions, potentially affects the Sertoli–SSCs’ network and consequently inhibits the SSCs’ self-renewal process.     

Ectopic Leydig Cells in Seminiferous Tubules of an Infertile Human Male with a Chromosomal Aberration

A case of a human male infertility with chromosomal aberration is reported. The patient showed neither mental retardation nor physical abnormalities except that the testes were somewhat small and soft. Plasma follicle stimulating hormone and luteinizing hormone were 49.0 and 19.0 mIU/ml. Plasma testosterone was 2.6 ng/ml. Karyotype was considered to be 46 XY q-, long arms of the Y chromosome being deleted. Histological features of the testis were peculiar. Seminiferous tubules were small and devoid of spermatogenic cells, consisting only of Sertoli cells. Peritubular boundary layer of the tubules showed a marked increase in width due to the increase of collagen fibers. The base of some Sertoli cells was seen to protrude into the thickened peritubular boundary layer or, though rare, into the interstitial space. Unusual cells which had a round vesicular nucleus and abundant, dense cytoplasms also occurred in the boundary layer of most tubules. These cells were identified as Leydig cells because of an extensively developed smooth endoplasmic reticulum in their cytoplasm, although they lacked Reinke's crystals. These ectopic Leydig cells sometimes lay in direct contact with Sertoli cells in the seminiferous tubule.     

Temperature and germ cell regulation of Leydig cell proliferation stimulated by Sertoli cell-secreted mitogenic factor: a possible role in cryptorchidism

Summary. Local control of Leydig cell morphology and function by seminiferous tubules was suggested in previous in vivo studies, especially those that used experimental cryptorchid rat testis as a model. These studies reported changes in morphology, increases in cell number and mitotic index and decreases in testosterone formation and luteinizing hormone/human chorionic gonadotropin receptor levels of Leydig cells. However, little is known about how these changes are mediated. We recently observed that a novel Sertoli cell-secreted mitogenic factor stimulated proliferation, decreased testosterone formation and luteinizing hormone/human chorionic gonadotropin receptor levels, and dramatically altered the morphology of Leydig cells in culture. In the present studies, we demonstrate that an increase in coculture temperature from 33 to 37 °C increased [³H]-thymidine incorporation (5.6- vs. 19.2-fold) and labelling index (4.3% vs. 15.8%), and accelerated proliferation (2.1- vs. 3.9-fold) of cultured immature Leydig cells. In addition, testosterone formation and luteinizing hormone/human chorionic gonadotropin receptor levels of Leydig cells cocultured with Sertoli cells were further decreased following a 4°C increase in coculture temperature. This elevation in culture temperature increased both the secretion of this factor by Sertoli cells and responsiveness of Leydig cells to this factor. In addition, the presence of germ cells, especially pachytene spermatocytes, inhibited the secretion of the mitogenic factor by Sertoli cells. These temperature- and germ cell-associated effects mimicked the morphological and functional changes of Leydig cells reported following experimental cryptorchidism. These observations suggest a possible role of this Sertoli cell-secreted mitogenic factor in explaining Leydig cell changes following experimental cryptorchidism.     

Is testosterone involved in the initiation of spermatogenesis in humans? A clinicopathological presentation and physiological considerations in four patients with Leydig cell tumours of the testis or secondary Leydig cell hyperplasia

The purpose of this study was to evaluate the role of testosterone on the puberal development of spermatogenesis and to present additional clinicopathological data which bring about new information to this controversial subject. Four pre-pubertal patients are presented, 2 of them bearing Leydig cell tumours of the testis in the form of nodular masses. In both cases seminiferous tubules in the immediate vecinity to the tumours showed complete development of spermatogenesis, while those located away from the tumours were infantile in nature. Gonadotrophic levels were within the normal pre-pubertal range in these 2 cases. In one of the patients, testosterone concentration in the testis showed higher values than normal, and a concentration gradient was detected between the tumoral nodule and non-tumoral parenchyma. The 3rd patient had a pineal choriocarcinoma producing high amounts of hCG and consequently a diffuse hyperplasia of Leydig cells with high levels of plasma testosterone. Seminiferous tubules showed development up to pachytene spermatocytes. The last case was a precocious puberty in a boy with a tumour of the 3rd ventricle area. He had elevated levels of testosterone in the testis and plasma. In the testicular biopsy, stimulation of Leydig cells was detected. The seminiferous tubules showed mature Sertoli cells and pachytene spermatocytes. FSH levels were abnormally low. These 4 cases present in common different situations in which abnormally high amounts of testos-happens in the immature rat, the interaction between testosterone and gonadotrophins is essential for the normal initiation of spermatogenesis in normal puberty. Considerations are discussed on the possible synergistic role of gonadotrophins or other factors in relation with stimulation of seminiferous tubules by testosterone.     

Regulation of interstitial cell function by seminiferous tubules in intact and cryptorchid rats

The effects of experimental cryptorchidism on seminiferous tubule secretions and interstitial cell testosterone production were studied in vitro. Spent media obtained from incubations of seminiferous tubules (SMST) from cryptorchid rats caused a significant increase in testosterone production when added to interstitial cells isolated from intact rats. The previously noticed inhibitory activity of the SMST from stages VIII–XI of the sperma-togenic epithelial cycle gradually disappeared after the induction of experimental cryptorchidism. SMST obtained from both sham-operated or cryptorchid rats stimulated basal testosterone production when added to interstitial cells from cryptrochid rats. SMST from rats had been cryptorchid for 7, 14 and 28 days stimulated testosterone production when added to interstitial cells prepared from intact animals. Seminiferous tubules from cryptorchid rats therefore appear to be the source of a heat stable, trypsin-resistant factor with an apparent molecular weight of between 5000 and 10 000 daltons which stimulates testosterone production when added to interstitial cells in vitro. Its activity could not be blocked by an LRH antagonist. This factor enhances both basal and LH-stimulated secretion of testosterone in contrast to the inhibitory activity which involves only a partial blockade of LH-dependent steroidogenesis.