Testicular Luteinizing Hormone Receptors Research Articles

Assessment of Testicular Lhcgr mRNA Expression Correlated with Testis and Seminal Vesicle Activities in the Libyan jird (Meriones libycus, Rodentia: Muridae) during Breeding Season Compared with Nonbreeding Season.

Simple SummaryThe breeding periods of desert rodents should be favorable to the survival of small young, conditioned by the availability of food that occurs in the Libyan jird biotope during the long photoperiod and the high temperatures. The Libyan jird (Meriones libycus) were caught in their natural biotope in the Saharan desert in Algeria and showed a seasonal cycle of the testis activity, characterized by the highest peak during spring (the breeding season) and the lowest activity was registered during autumn and winter (nonbreeding season). Lhcgr mRNA expression is increased in autumn and decreased in spring. This expression varied in an opposite manner to testicular and seminal vesicle structures. The Libyan jird (Meriones libycus, 1823) is a wild desert rodent that is a seasonal breeder species adapted to breed when the environmental conditions can satisfy the energy and hydrous requirements of pregnant and nursing females to ensure that births occur at the most favorable time of the year. We assessed gene expression of testicular luteinizing hormone receptor (Lhcgr) correlated to testis activity. The expression of Lhcgr was evaluated using quantitative Real Time-Polymerase Chain Reaction (qRT-PCR and the testis activity by a histological method in adult male Libyan jirds during the nonbreeding and breeding seasons. Our results showed that Lhcgr mRNA expression increased in autumn during the nonbreeding season and decreased in spring during the breeding season. This expression varied in contrast to testicular structure or function and plasma testosterone levels. These results help to elucidate this desert rodent’s seasonal sexual activity, which is correlated with central regulation.

Seasonally sympatric but allochronic: differential expression of hypothalamic genes in a songbird during gonadal development.

Allochrony, the mismatch of reproductive schedules, is one mechanism that can mediate sympatric speciation and diversification. In songbirds, the transition into breeding condition and gonadal growth is regulated by the hypothalamic-pituitary-gonadal (HPG) axis at multiple levels. We investigated whether the difference in reproductive timing between two seasonally sympatric subspecies of dark-eyed juncos (Junco hyemalis) was related to gene expression along the HPG axis. During the sympatric pre-breeding stage, we measured hypothalamic and testicular mRNA expression of candidate genes via qPCR in captive male juncos. For hypothalamic mRNA, we found our earlier breeding subspecies had increased expression of gonadotropin-releasing hormone (GnRH) and decreased expression of androgen receptor, oestrogen receptor alpha and mineralocorticoid receptor (MR). Subspecies did not differ in expression of hypothalamic gonadotropin-inhibitory hormone (GnIH) and glucocorticoid receptor (GR). While our earlier breeding subspecies had higher mRNA expression of testicular GR, subspecies did not differ in testicular luteinizing hormone receptor, follicle-stimulating hormone receptor or MR mRNA expression levels. Our findings indicate increased GnRH production and decreased hypothalamic sensitivity to sex steroid negative feedback as factors promoting differences in the timing of gonadal recrudescence between recently diverged populations. Differential gene expression along the HPG axis may facilitate species diversification under seasonal sympatry.

Di-(2-ethylhexyl)-phthalate disrupts pituitary and testicular hormonal functions to reduce sperm quality in mature goldfish

Di-(2-ethylhexyl) phthalate (DEHP) interferes with male reproductive endocrine system in mammals, however its effects on fish reproduction are largely unknown. We evaluated sperm quality and investigated reproductive endocrine system in mature goldfish (Carassius auratus) exposed to nominal 1, 10, and 100μg/L DEHP. To examine DEHP estrogenic activity, one group of goldfish was exposed to 17β-estradiol (5μg/L E2) for comparison. Following 30d of exposure, sperm production was decreased and suppressed in DEHP and E2 treated goldfish, respectively. Sperm motility and velocity were decreased in goldfish exposed to 100 and 10μg/L DEHP at 15s post-sperm activation, respectively. Compared to control, 11-ketotestosterone (11-KT) levels were decreased at 10 and 1μg/L DEHP at day 15 and 30, respectively. In E2 treated goldfish, 11-KT levels were decreased compared to control during the period of exposure. E2 levels were increased in goldfish exposed to E2, but remained unchanged in DEHP treated goldfish during the period of exposure. StAR mRNA levels encoding regulator of cholesterol transfer to steroidogenesis were decreased in DEHP and E2 treated goldfish following 15 and 30d of exposure, respectively. Luteinizing hormone (LH) levels were decreased in DEHP and E2 treated goldfish following 15 and 30d of exposure, respectively. In DEHP treated goldfish, gnrh3, kiss1 and its receptor (gpr54) mRNA levels did not change during the experimental period. In E2 treated goldfish, gnrh3 mRNA levels were decreased at day 7, but kiss1 and gpr54 mRNA levels were increased at day 30 of exposure. The mRNA levels of genes encoding testicular LH and androgen receptors remained unchanged in DEHP and E2 treated goldfish. In contrast to E2 treated goldfish, vitellogenin production was not induced in DEHP treated goldfish and mRNA levels of genes with products mediating estrogenic effects remained unchanged or decreased. In conclusion, DEHP interferes with testis and pituitary hormonal functions to reduce sperm quality in goldfish and does not exhibit estrogenic activity.

The effect of active immunization against vasoactive intestinal peptide and inhibin on reproductive performance of young White Leghorn roosters

The neuropeptides vasoactive intestinal peptide (VIP) and gonadal inhibin have long been considered putative regulators of reproduction in hens. However, their role in young roosters remains unclear. We studied the effect of active immunization against VIP, inhibin, and a combination of both hormones on reproduction in young White Leghorn roosters. At 13 wk of age, White Leghorn roosters (n = 60) were split into 4 groups (n = 15). One group was actively immunized against VIP, the second against inhibin, the third against both VIP and inhibin, and the fourth, untreated, served as a control. Active immunization against VIP enhanced reproductive parameters as manifested by increased semen quality, plasma steroid levels, and mRNA gene expression of hypothalamic gonadotropin-releasing hormone-I, pituitary follicle-stimulating hormone, pituitary luteinizing hormone (LH), and decreased mRNA gene expression of hypothalamic VIP, pituitary prolactin, and testicular LH receptor. In contrast, immunization against inhibin decreased reproductive parameters such as semen quality, plasma steroid levels, mRNA gene expression of pituitary follicle-stimulating hormone and testicular inhibin. The combined treatment showed the greatest increase in semen quality parameters, plasma steroid levels, and mRNA gene expression of hypothalamic gonadotropin-releasing hormone-I, pituitary follicle-stimulating hormone, pituitary LH, and testicular LH receptor. Moreover, it significantly reduced mRNA gene expression of hypothalamic VIP and pituitary prolactin and mildly reduced that of testicular inhibin. These results suggest that VIP plays a negative role, at a young age, in reproduction of roosters that is similar to that in hens and that inhibin is as important in reproductive function in young roosters as in mammals.

Effects of chronic bhang (cannabis) administration on the reproductive system of male mice

The purpose of this study was to investigate the effect of chronic uptake of bhang, prepared from the Cannabis sativa, on male reproductive physiology in adult male Parkes strain (P) mice. An attempt was also made to investigate the presence of cannabinoid 1 (CB1) and cannabinoid 2 (CB2) receptors, and fatty acid amide hydrolase (FAAH) in the testis and to evaluate any changes in it resulting from chronic intake of bhang in mice. Adult male mice were given bhang (3 or 6 mg/kg body weight/day) orally for 36 consecutive days. Chronic intake of bhang caused regressive changes in the testes and suppressed sperm count, viability and motility. Bhang intake also caused significant decline in circulating testosterone level due to decline in testicular 3β HSD enzyme activity. An immunohistochemical study demonstrated the presence of CB1, CB2 and FAAH in the testis of mice. The present study also showed significant variation in the CB1 and CB2 receptors and FAAH protein levels in testes of mice exposed to bhang. These suppressive effects may be due to inhibitory effect of bhang on pituitary expression of gonadotrophin releasing hormone (GnRH) I receptor protein. Treatment of testes with bhang in vitro significantly decreased testicular luteinizing hormone receptor (LHR) and FAAH expression suggesting direct action of bhang on testicular activity. The findings of this study thus suggest that bhang may impair fertility in male mice through alteration in the testicular endocannabinoid system and that chronic bhang exposure in humans would be predicted to alter male fertility.

Effects of exogenous testosterone on testicular luteinizing hormone and follicle-stimulating hormone receptors during aging.

During aging, the male Japanese quail exhibits a loss of fertility, increased morphological abnormalities in the testes, and a higher incidence of Sertoli cell tumors. Although there is a coincident loss of reproductive behavior, plasma androgen levels remain high until testicular regression occurs in association with senescence. The purpose of this study was to compare mean specific binding of chicken luteinizing hormone (LH) and follicle-stimulating hormone (FSH) as a measure of testicular receptors during identified stages during aging. Males were categorized according to age (young = 9 months, middle aged = 24 months, or old = 36+ months) and sexual behavior (active or inactive). Testicular samples were collected immediately after perfusion with 4% paraformaldehyde from the following groups: young active (n = 8), young photoregressed (n = 5), young photoregressed plus testosterone implant (n = 4), middle-aged active (n = 8), middle-aged inactive (n = 4), old inactive (n = 5), and old inactive plus testosterone implant (n = 6). A crude plasma membrane fraction was prepared from the testes of each bird and an aliquot deriving from 10 mg of testicular tissue was used for binding assay. Specific binding of labeled LH or FSH was expressed as percentage of total radioactive hormone. Results showed significant (P < 0.05) age-related decreases in both FSH and LH receptor numbers. The highest FSH binding was found in young and middle-aged active males, with low binding in old inactive males. Testicular LH binding decreased during aging, with a sharp decrease in middle-aged males, which was similar to old males. Testosterone implants weakly stimulated FSH and LH binding in old males. Both LH and FSH binding decreased in photoregressed young males. However, testosterone implants stimulated increased LH binding, but did not affect FSH binding in young photoregressed males. These results provide evidence for separate regulation of testicular LH and FSH receptors, with testosterone stimulation of LH receptor, but not FSH receptor number in young males. However, during aging there appears to be a loss of this response, potentially because of the reduced efficacy of testosterone stimulation, thereby implying a diminished capacity for response with aging.

Cloning and sequence analysis of the extracellular region of the polar bear (Ursus maritimus) luteinizing hormone receptor (LHr), follicle stimulating hormone receptor (FSHr), and prolactin receptor (PRLr) genes and their expression in the testis of the black bear (Ursus americanus)

Male black bears undergo seasonal changes in testicular activity. The testes are fully functional from May through July, regress from July through December, and recrudesce from January until May. The mechanisms responsible for the initiation of testicular recrudescence in the bear are unknown. The objectives of this study were to: (1) clone and sequence a substantial fragment of the extracellular portion of the luteinizing hormone receptor (LHr: 646 bp) and follicle stimulating hormone receptor (FSHr: 852 bp), and the extracellular/transmembrane portion of the prolactin receptor (PRLr: 680 bp) in the bear using reverse transcription-polymerase chain reaction (RT-PCR); and (2) determine whether the expression of LH-, FSH-, and PRL-receptor mRNA transcripts differs between the beginning and terminal stages of testicular recrudescence. Comparisons of the partial cDNA and predicted amino acid sequences of ursine receptors with the corresponding sequences from the pig, cow, human, and rat suggest that the LHr and FSHr are highly conserved (LHr: 87.1-93.7%; FSHr: 86.0-92.7%) whereas the PRLr is less well conserved (81-87%). Testicular LHr mRNA was more abundant during the breeding season in May than during the non-breeding season (early stage of recrudescence) in January. In contrast, testicular FSHr mRNA abundance was greater in January than in May. Testicular PRLr mRNA appeared equally abundant in January and May; however, two additional transcripts were present during the breeding season in May. This study provides molecular tools for future investigations of the control of testicular recrudescence in the black bear and demonstrates that the expression of testicular gonadotropin and PRL receptor mRNA is seasonally regulated. Mol. Reprod. Dev. 55:136-145, 2000.

HCG binding to the testicular LH receptor is similar in fertile, subfertile, and infertile stallions.

Recent evidence in our laboratory suggests that the cause of idiopathic subfertility/infertility in breeding stallions may originate in the testes at the luteinizing hormone (LH) receptor or postreceptor level. The objective of this research was to determine if LH receptor binding activity is altered in subfertile and infertile stallions. Six fertile, three subfertile, and three infertile stallions, ages 11-23 years, were classified according to normal semen parameters and pregnancy rates and then castrated in the breeding season. Blood was collected prior to castration, and plasma was stored until analyzed for LH, follicle stimulating hormone (FSH), estrogen conjugates (EC), estradiol (E2), testosterone (T), and inhibin (I) by radioimmunoassay (RIA). Testicular cell membranes were prepared and snap-frozen until analyzed for LH binding activity by radioreceptorassay (RRA) using increasing amounts of I125 human chorionic gonadotropin (hCG). Luteinizing hormone receptor numbers and affinity constants were determined by Scatchard analysis. Plasma LH, FSH, EC, E2, and T levels did not differ between fertile and subfertile stallions, but LH and FSH were significantly higher (P < 0.05) and EC, E2, T, and I levels were significantly lower (P < 0.05) in infertile stallions as compared to fertile and subfertile stallions. Receptor number (Rt) and affinity constants Ka were similar (P > 0.05) between fertile (Rt = 9.44 x 10(-11) M, Ka = 0.300 x 10(10) M-1), subfertile (Rt = 13.02 x 10(-11) M, Ka = 0.194 x 10(10) M-1), and infertile (Rt = 7.65 x 10(-11) M, Ka = 0.380 x 10(10) M-1) stallions. In conclusion, these data suggest that an endocrine dysfunction in the testes of stallions with poor fertility may not be due to a LH receptor disorder but may be due to a postreceptor malfunction.

Expression of Human Luteinizing Hormone (LH) Receptor: Interaction with LH and Chorionic Gonadotropin from Human but not Equine, Rat, and Ovine Species

Studies on human LH receptors are difficult due to the limited availability of clinical samples. Recent cloning of rat and porcine LH receptor cDNAs indicated that these binding sites are single polypeptides of the G-protein-coupled receptor family with seven transmembrane domains. Based on the conserved sequences of rat and porcine receptors, we performed reverse transcription polymerase chain reaction, using human ovarian mRNA as template and obtained partial human LH receptor cDNA clones. Further screening of a human ovary cDNA library and subsequent ligation of individual cDNA clones generated a human LH receptor cDNA containing the entire amino acid-coding region. Sequence analysis indicated that the human receptor cDNA displays 89% and 82% homology at the nucleotide level with its porcine and rat counterparts, respectively. A region spanning the second extracellular and third transmembrane domains is highly conserved among the human LH, FSH, and TSH receptors. The ovarian LH receptor clone is, however, significantly different from an incompletely spliced LH receptor cDNA recently obtained from a human thyroid library. Unlike the thyroid clone, the ovarian LH receptor cDNA could be expressed in the human fetal kidney cell line (293), and radioligand receptor assay identified high affinity (Kd, 1.2 x 10(-10) M) LH/hCG-binding sites on the plasma membrane. Binding specificity of the human LH receptor was studied using recombinant human CG, LH, and FSH secreted by CHO cells transfected with the respective genes. Human CG and LH displaced [125I]hCG binding with an ED50 of 4.3 and 4.8 ng/ml, respectively. In contrast, recombinant FSH was not effective. Treatment of transfected cells with recombinant gonadotropins also induced dose-dependent increases in extracellular cAMP production (hCG = LH much greater than FSH; ED50 25, 10, and greater than 3000 ng/ml). Although equine, rat, and ovine LH as well as equine CG competed effectively for rat testicular LH receptor binding, these hormones were unable to displace [125I]hCG binding to the human receptor, suggesting evolutionary changes in receptor binding specificity and the importance of using human receptors for clinical studies. Thus, the cloning and expression of the human LH receptor cDNA allowed analysis of interactions between human LH receptor and gonadotropins from diverse species. The present work should provide the basis for future design of therapeutic agents capable of interacting with the human receptor and for understanding the structural basis for LH receptor binding to different gonadotropins.

Growth Hormone and Insulin-Like Growth Factor I Treatment Increase Testicular Luteinizing Hormone Receptors and Steroidogenic Responsiveness of Growth Hormone Deficient Dwarf Mice*

To test the hypothesis that insulin-like growth factor (IGF-I) is required for the in vivo development of testicular Leydig cell function, either recombinant human GH [(hGH) (1.5 micrograms/g BW) or recombinant IGF-I (1 microgram/g BW) was injected three times daily into immature Snell dwarf mice (dw/dw) and into phenotypically normal control (Dw/-) for 7 days. In dw/dw mice hGH enhanced significantly body, liver, kidney, and testicular weight. In addition, hGH increased testicular LH receptors and the acute steroidogenic response to human CG, but there was no significant effect on basal plasma testosterone or plasma LH levels. The effects of IGF-I in body and kidney weight were less pronounced than those produced by hGH, but its effects on testicular weight and LH receptors, as well as on the acute steroidogenic response to human CG, were similar to that observed after hGH treatment. In Dw/- mice hGH had no effect on either body or organ weight or on testicular function, despite the fact that it induced a significant increase in plasma IGF-I levels. These results indicated that IGF-I is able to induce the maturation of Leydig cell function and that the effects of hGH on the testis are probably mediated by IGF-I. They also suggest that the delayed puberty associated with GH deficiency or resistance is most likely related to an IGF-I deficiency.

Effects of prolactin and bromocryptine on the regulation of testicular luteinizing hormone receptors in mice

The binding of luteinizing hormone (LH) to testicular homogenates increased gradually in mice from 15 to 60 days of age, while the level at 90 days was almost the same as that at 60 days. The plasma concentration of prolactin (PRL) increased significantly from 15 to 40 days and thereafter remained constant. In order to ascertain the influence of PRL on testicular receptors for LH, bromocryptine was injected subcutaneously for 10 days into immature (20-day-old) and adult (90-day-old) mice. In 20-day-old mice, treatment with bromocryptine significantly reduced the plasma levels of PRL but had no significant effects on the binding of LH to receptors in 30-day-old mice. However, in 90-day-old mice, treatment with bromocryptine led to a significant reduction in numbers of receptors for LH 10 days later. There was no difference in dissociation constants (Kd) between groups of oil-injected (Kd = 6.5 x 10(-10) M) and bromocryptine-injected (Kd = 4.6 x 10(-10) M) mice. The reduction in binding of LH per testis of 100-day-old mice after treatment with bromocryptine was eliminated by the simultaneous administration of ovine (o) PRL. The plasma level of follicle-stimulating hormone (FSH) in 100-day-old mice, which tended to be decreased as a result of treatment with bromocryptine, was markedly increased by treatment with oPRL. There were no distinct changes in binding of FSH in any of the groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Pituitary and gonadal function during physical exercise in the male rat

The effects of training and acute exercise on serum testosterone, luteinizing hormone (LH) and corticosterone levels and on testicular endocrine function in male rats were studied. In the first part of the study, the rats were trained progressively on a treadmill, over 8 weeks. Training did not change the basal levels of serum testosterone, LH and corticosterone, or the testicular concentrations of testosterone and its precursors progesterone and androstenedione. The levels of testicular LH (30.3 ± 2.6 ng/g wet wt, mean ± SEM) and lactogen (150± 14 pg/g) receptors were unchanged after training. However, the capacity of testicular interstitial cell suspensions to produce cAMP and testosterone increased by 20–30% during in vitro gonadotropin stimulation.In the second part, the trained and untrained control animals underwent acute exhaustive exercise. Serum testosterone levels decreased by 74 and 42% in trained and untrained rats, respectively (P < 0.02), and corticosterone rose by 182% in trained and 146% in untrained rats (P < 0.01), whereas the LH level was unchanged. Testicular levels of testosterone and its precursors decreased, with the exception of unchanged androstenedione, in trained rats; the cAMP concentration was unchanged. In both trained and untrained rats, acute exercise decreased the capacity of interstitial cell suspensions to produce cAMP, whereas there were no consistent effects on testosterone production. Acute exercise had no effect on LH or lactogen receptors in testis tissue.In conclusion, training had no effect on serum or testicular androgen concentrations, but increased Leydig cell capacity to produce testosterone and cAMP. Acute exercise decreased serum and testicular testosterone concentrations without affecting serum LH. A direct inhibitory effect of the increased serum corticosterone level on the hypothalamic-pituitary level and/or testis may be the explanation for this finding.

Changes in the testicular binding of luteinizing hormone and plasma testosterone concentrations in the Djungarian hamster subjected to different photoperiods and temperatures and effects of long-term testosterone treatment on the binding.

The effects of artificial photoperiod, temperature, and long-term testosterone treatment on testicular luteinizing hormone (LH) binding were studied in adult male Djungarian hamsters. In hamsters transferred to long-day (LD; 16 hr light, 8 hr dark) photoperiod 8 weeks after adaptation in short-day (SD; 8 hr light, 16 hr dark) photoperiod of 25 degrees C, testicular growth was associated with an increase in the total LH binding per two testes and a decrease in LH binding per unit testicular weight. Plasma testosterone levels reached a peak 47 days after transfer to LD and tended to decrease thereafter, while the testes continued growing. In contrast, when hamsters reared under LD conditions at 25 degrees C for 12 weeks were transferred to SD, testicular regression was associated with a decrease in plasma testosterone and the total LH binding per two testes and an increase in LH binding per unit testicular weight. A significant decrease in LH binding per unit weight compared to SD controls was observed in those hamsters exposed to SD with continuous testosterone treatment. The testosterone treatment tended to induce decrease in the total LH binding. Scatchard plot analyses of the binding suggested that changes in LH binding were due to changes in the number of binding sites. When sexually mature male hamsters were subjected for 8 weeks to two different ambient temperatures (7 degrees C and 25 degrees C) and photoperiods (LD and SD), the difference between the two temperature groups was statistically not significant regarding the weights of testes, epididymides, and prostates; plasma testosterone levels; and LH binding in either LD or SD group. These results suggest that photoperiod is a more important environmental factor than temperature for the regulation of testicular activity and LH receptors and that testosterone reduces the number of LH receptors per unit testicular weight in adult male Djungarian hamsters.

SDZ 200–110 Induces Leydig Cell Tumors By Increasing Gonadotropins in Rats

The administration of 62.5 mg/kg/day of SDZ 200–110, a calcium channel blocker, for 2 years increased the incidence of Leydig cell tumors while decreasing pituitary tumors in Sprague-Dawley rats. Lower doses did not change the incidence of these tumors. No other endocrine tumors were seen in rats or mice of either sex. A single gavage dose of 62.5 mg/kg/day decreased serum testosterone levels by 90% 4 hr after dosing. In vitro testosterone production by Leydig cells from these animals was minimally decreased, which suggests that a direct inhibition of steroid synthesis was removed during cell isolation. Dietary administration of the drug for 10 weeks did not significantly alter levels of serum hormones or testicular luteinizing hormone (LH) and gonadotropin-releasing hormone (GnRH) receptors, although a significant elevation of testicular testosterone levels was seen. Increased serum levels of LH and follicle-stimulating hormone (FSH) were seen after 52 and 66 weeks, respectively, of dietary feeding of 62.5 mg/kg/day. The increase in serum LH was observed to week 104, while FSH levels returned to control levels by week 94. No effect on gonadotropin receptors was seen at the 6.25 mg/kg/day dosage. The age-related increase in serum prolactin was markedly reduced by 62.5 mg/kg/day of SDZ 200–110 in weeks 66 to 104 and to a lesser extent at the 6.25 mg/kg/day dosage. Testicular LH receptors were decreased by the high dose in animals sacrificed after 90–104 weeks. In conclusion, SDZ 200–110 increases the incidence of Leydig cell tumors by elevating levels of serum gonadotropins. The suggested mechanism for this increase in gonadotropins is a result of the effects of SDZ 200–110 on serum hormones and testicular LH receptors. The drug was judged not to pose a risk to humans since no change in gonadotropin levels was observed after chronic treatment.

Short-term pituitary-testicular responses of prepubertal ram lambs to hemicastration.

To examine the short-term effects of hemicastration on pituitary-gonadal responses, 12 ram lambs were anesthetized and hemicastrated at 4 mo of age and killed (n = 4) at 2 (HC2), 7 (HC7), or 14 (HC14) days following surgery. Four intact (INT) rams were killed 14 days following anesthesia. Testis and pituitary weights were similar between HC and INT rams. Serum follicle-stimulating hormone (FSH) in HC rams increased within 6 h, peaked at 12 h, and remained elevated above INT levels throughout the study. Overall mean serum testosterone levels in HC rams were lower than in INT rams for the first 48 h, but were similar by 3 days post-surgery. Pulsatile luteinizing hormone (LH) and testosterone secretion was suppressed for the first 9.5 h following anesthesia and/or surgery in both HC and INT animals. A single LH pulse and succeeding testosterone pulse occurred in 10/12 HC and 4/4 INT rams between 10 and 14 h post-surgery, both of which were lower in amplitude in HC than INT animals. However, on Day 7, pulsatile secretory patterns of LH and testosterone were similar, suggesting compensatory androgen secretion had occurred in HC rams. Pituitary LH content was unaffected by hemicastration. In contrast, pituitary FSH content was greater in HC7 and HC14 compared to HC2 and INT animals. Pituitary gonadotropin hormone-releasing hormone (GnRH) receptor concentrations were similar in INT, HC7, and HC14 rams, but were slightly reduced in HC2 rams. Neither testicular LH nor FSH receptor concentrations were altered by hemicastration at any time.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of castration or testosterone implants upon pituitary function in hypogonadal mice bearing normal foetal preoptic area grafts.

Grafts of normal mouse preoptic area (POA) tissue into the third ventricle of gonadotrophin-releasing hormone (GnRH)-deficient hypogonadal (hpg) mice resulted in an elevation of pituitary GnRH receptors, an increased synthesis of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) by the pituitary gland, an elevation of gonadal LH receptors and in the stimulation of steroidogenesis and spermatogenesis in the testis. In normal mice both castration or the subcutaneous implantation of testosterone capsules for 10 days reduced GnRH receptors, pituitary LH and FSH content, and the latter treatment also caused a 50% reduction in testicular LH receptors. In hpg mice bearing POA grafts testosterone implants failed to affect any of the above parameters, and castration failed to affect pituitary gonadotrophin hormone content, although there was a slight reduction in pituitary GnRH receptors after castration. These experiments suggest that neither the pituitary gonadotroph, nor the GnRH neurone represent major sites for the direct negative feedback of testosterone upon gonadotrophic hormone secretion in male mice.

Seasonal differences in testicular receptors and steroidogenesis

Gonadal function in most animal species exhibits considerable annual fluctuations, with gametogenesis and fertility often being confined to a short and rigidly controlled breeding season. In males, production of androgenic steroids by the testis is usually maximal immediately before and during the breeding season. In the golden hamster, seasonal regression of the testes is associated with decrease in the total content of luteinizing hormone (LH), follicle-stimulating hormone (FSH) and prolactin (PRL) receptors, and similar findings have been reported for other mammalian species. However, the concentration of LH and PSH receptors per unit of testis weight is typically elevated rather than suppressed during testicular regression. Reduction in the number of testicular LH and PRL receptors in adult golden hamsters exposed to short photoperiod is due primarily to suppression of pituitary PRL release under these circumstances. Regulation of seasonal changes in testicular FSH binding, as well as regulation of the levels of LH, PRL and FSH receptors in other seasonally breeding species remain to be elucidated. Reduction in the content of LH receptors in the testes is accompanied by reduced capacity to produce androgens in response to LH stimulation. Although these events are likely to be causally related, other mechanisms are also involved. In particular, seasonal regression is accompanied by reduced capacity of the testes to convert C 21 steroid precursors into biologically active androgens. Seasonal loss of FSH receptors was reported to be accompanied by increased rather than reduced responsiveness of the Sertoli cells to FSH, thus resembling the situation in immature animals. It can be concluded that alterations in the ability of the testes to bind pituitary gonadotropins and to respond to gonadotropic stimulation are among the mechanisms responsible for seasonal shifts between gonadal activity and quiescence.

Specific gonadotrophin binding sites in the bullfrog testis

We demonstrated the presence of specific binding sites for bullfrog luteinizing hormone (LH) and follicle-stimulating hormone (FSH) in a testicular crude plasma membrane fraction of the bullfrog, Rana catesbeiana. The equilibrium analysis of the binding showed that a straight line was fitted to the Scatchard plot of LH and a curvilinear line to that of FSH, suggesting the presence of single-type binding sites for LH and the presence of multiple-type binding sites or negative cooperactivity of the binding for FSH. Competition experiments showed that bullfrog LH could replace the specific binding of bullfrog FSH and vice versa. Binding affinity of bullfrog LH to radioiodinated bullfrog FSH binding sites was as high as that of intact bullfrog FSH to the same sites. In contrast, binding properties of bullfrog FSH to radioiodinated bullfrog LH binding sites were not simple: the competition curve obtained with FSH against the binding of bullfrog LH had an extremely low slope value and was not parallel to that obtained with LH. These results suggest that differentiation of LH and FSH receptors is not complete in this species, although it has been reported that these hormones have separate actions on the testis. Competition experiments further showed that bullfrog testicular LH and FSH receptors possessed higher affinities for gonadotrophins of homologous or closely related animal species compared with phylogenically distant groups. Species specificity of the hormone-receptor interaction may cause species specificity of the gonadotrophin action.

Elevation of serum testosterone during chronic LHRH agonist treatment in the bull.

The objective of this study was to try to depress serum testosterone (T) in bulls by prolonged treatment with a potent luteinizing hormone-releasing hormone (LHRH) agonist. Eight sexually mature bulls (325 to 475 kg) were assigned to treatment or control groups. Treatment consisted of 150 micrograms nafarelin acetate 6-D-2-naphthyl-alanine-LHRH (LHRH-A) injected im every 6 h for 15 d. Bovine serum albumin (BSA, .01%) in a carrier solution was injected at the same times in control bulls. Serial 15-min blood samples were collected via jugular cannula during the initial 36 h of treatment and during 6-h windows on d 4, 8 and 14. Bulls were slaughtered and pituitaries and testes collected on d 15. Serum luteinizing hormone (LH), follicle stimulating hormone (FSH) and T were elevated after initial injection of LHRH-A, but returned to basal concentrations by 12, 5 and 17 h, respectively. Prolonged LHRH-A treatment prevented pulsatile LH and T secretion compared with control bulls. Mean serum LH did not differ from that of controls on d 4, 8 and 14 of LHRH-A treatment, while serum T was elevated (P less than .01) during the same time periods. Oscillating patterns and mean concentrations of serum FSH were not different between control and LHRH-A-treated bulls. Fifteen days of LHRH-A treatment depressed pituitary LHRH receptor numbers (P less than .05) and pituitary LH (P less than .01) and FSH (P less than .05) concentrations. Testicular LH receptor numbers were elevated (P less than .01), but testicular FSH receptor numbers were not altered.(ABSTRACT TRUNCATED AT 250 WORDS)

Autoregulation of testicular luteinizing hormone receptors in hypogonadal (hpg/hpg) mice.

The autoregulation of testicular luteinizing hormone (LH) receptors was studied in hypogonadal (hpg/hpg) and normal mice. The basal concentration of LH receptors was more than three-fold higher in hpg/hpg than in normal mice. After injection of hCG, hpg/hpg mice showed a decrease in LH receptor levels which was not observed in normal mice. Plasma testosterone was undetectable in hpg/hpg mice, even after treatment with a single dose of hCG. Plasma prolactin levels were higher in hpg/hpg than in normal mice. The increase in basal LH receptor levels is thought to be due to a compensatory mechanism in which elevated prolactin could play a role. The differences between hpg/hpg and normal mice in the autoregulation of LH receptors observed could be due to the "hypersensitivity" of the physiologically immature testis in hpg/hpg mice to the action of hCG, to gonadotropin deficiency, particularly during the earlier stages of development, or to a direct effect of the hpg locus on the metabolism of LH receptors.