Number Of Luteinizing Hormone Receptors Research Articles

Enough is enough! Patients who do not conceive on 600 IU/d of gonadotropins show no improvement from an additional 150 IU of LH activity

Studies have suggested that supplemental LH improves outcomes in assisted reproductive technology (ART) cycles. In this retrospective review, an additional 150 IU of LH activity did not improve ART outcomes in women undergoing a second round of IVF/ intracytoplasmic sperm injection (ICSI) after an initial failed cycle using 600 IU of gonadotropins.

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.

Breaking Up the T Party: Short-circuited hormone signal keeps old rats from making testosterone (Sex hormones)

Women haven't cornered the market on hormonal woes. As men age, their blood testosterone concentrations decline, which might contribute to health problems such as osteoporosis (see "More Than a Hot Flash" ). A new study casts blame on a withered signaling pathway that normally jolts testosterone-producing cells into action. Scientists don't understand what triggers the fall in testosterone concentration, but the Brown Norway rat, which experiences an ebb in testosterone concentration similar to that in humans, has provided some clues. In old animals, testosterone factories in the testes called Leydig cells lose their capacity to churn out the hormone. Previous studies indicated that supplemental luteinizing hormone (LH), which normally prods Leydig cells into action, couldn't elevate testosterone concentrations in old rats. To find out why, Chen and colleagues removed Leydig cells from young and old rats and cultured them in the presence of LH. Cells from the youngsters pumped out testosterone, but those from geezer rodents did not. To investigate why old cells fail to respond to LH, the team measured how much LH bound to each type of cell. Old cells glommed onto one-third fewer hormone molecules than young cells did, indicating a decrease in the number of cell surface receptors that bind LH. But a lack of receptors didn't cause the decline in testosterone production, according to the outcome of an additional experiment. The researchers collected Leydig cells from young animals whose LH output had been artificially lowered. These cells had fewer LH receptors--yet they made much more testosterone in culture than old cells did, a result that takes the number of LH receptors off the hook. Perhaps the pathway through which the LH receptor stimulates testosterone production is defective, the researchers proposed. After binding LH, the receptor triggers synthesis of a molecule called cyclic AMP (cAMP) inside Leydig cells; cAMP ignites testosterone production. Chen and colleagues found that cells from old rats contained lower cAMP concentrations than did those from either normal young animals or young animals with reduced LH. A general slump in cAMP formation wasn't to blame: Old cells pumped out the molecule when treated with a chemical that directly stimulates the cAMP-producing enzyme. The results suggest that aging short-circuits the LH receptor's ability to trigger cAMP synthesis in Leydig cells. How this glitch arises is mysterious, but the authors propose that Leydig cells might accrue oxidative damage while making testosterone. Enzymes generate reactive oxygen species as they manufacture testosterone, and previous work by the team suggests that decreasing testosterone output reduces oxidative damage in Leydig cells. The study provides new evidence for how rat Leydig cells lose their capacity to generate testosterone, says endocrinologist Alvin Matsumoto of the University of Washington, Seattle, but it's uncertain whether humans suffer from the same defect. The idea that free radicals assault Leydig cells is an intriguing one, although not proven, he says. If that's true, antioxidants might help maintain testosterone production. Perhaps the delicate broccoli flower will be the secret to youthful vigor in older men. --R. John Davenport; suggested by James M. Harper H. Chen, M. P. Hardy, B. R. Zirkin, Age-related decreases in Leydig cell testosterone production are not restored by exposure to LH in vitro. Endocrinology 143 , 1637-1642 (2002). [Abstract] [Full Text]

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.

Basic fibroblast growth factor induces luteinizing hormone receptor expression in the presence of insulin-like growth factor-I in ovarian granulosa cells

Effect of basic fibroblast growth factor (bFGF) on the expression of receptors for luteinizing hormone (LH), a marker of differentiation, was studied using estrogen-primed rat ovarian granulosa cells in primary culture. bFGF had no effect by itself but dose-dependently induced expression of functional LH receptors in the presence of insulin-like growth factor-I (IGF-I). The effect of a combination of bFGF and IGF-I was delayed in onset and the magnitude of the response was smaller when compared to the action of follicle-stimulating hormone (FSH). Scatchard analysis revealed that disspciation constant ( K d) and number of LH receptors induced by bFGF and IGF-I were 0.47 nM and 6.48 fmol/10 6 cells, respectively. Unlike FSH, bFGF plus IGF-I did not cause an immediate increase in cAMP release, however, considerable amount of cAMP release was observed in cells incubated for 72 h with bFGF plus IGF-I. Indomethacin, an inhibitor of cyclooxygenase, attenuated both LH receptor expression and cAMP release induced by bFGF plus IGF-I but had little effect on the action of FSH. Finally, a combination of bFGF and IGF-I increased production of prostaglandin E 2 in granulosa cells. These results indicate that bFGF is capable of inducing LH receptor in the presence of IGF-I by a mechanism involving production of prostaglandin E 2.

Effects of cortisol on the amounts of estradiol-17β and progesterone secreted and the number of luteinizing hormone receptors in cultured bovine granulosa cells

The direct effects of cortisol upon the amounts of estradiol-17β and progesterone secreted, and the number of luteinizing hormone (LH) receptors in cultured bovine granulosa cells were examined. Granulosa cells from small bovine follicles (diameter 3–7 mm) were successfully cultivated in a 1:1 mixture of Dulbecco's modified Eagle's medium and Ham's F-12 nutrient medium containing 10% fetal calf serum for the first day and subsequently incubated with follicle stimulating hormone and testosterone in serum-free medium for up to 8 days. The cells increased their output of both estradiol and progesterone during 9 days of culture. The addition of cortisol in the range 0.1–10 μM did not significantly change the total DNA content of the granulosa cells. Cortisol treatment caused a significant decrease in the amount of estradiol-17β secreted by granulosa cells ( P<0.05). Cortisol also significantly decreased the number of LH receptors in the cultured granulosa cells in a dose-dependent manner ( P<0.05). In contrast, cortisol tended to increase progesterone production by granulosa cells. These results demonstrate the direct inhibitory effect of cortisol on follicular secretion of estradiol-17β and LH receptor content. This observation is consistent with the hypothesis that cortisol released by stress may directly inhibit the functional maturation of the bovine follicles.

Progesterone regulation of luteinizing hormone receptors on cultured bovine luteal cells

During development of the corpus luteum (CL), the numbers of luteinizing hormone (LH) receptors increase. Cultured bovine luteal cells from developing and mature CL were used to examine the influence of progesterone (P 4) on this receptor. CL were obtained from dairy cows during the early or middle phase of the estrous cycle. In early CL, the number of receptors per cell was increased by exogenous progesterone treatment but there was no effect on receptor numbers in cells from midcycle CL. Binding affinities did not change with respect to age or treatment. Forskolin elevated endogenous progesterone and also enlarged the receptor population. The action did not appear to be an unmasking of cryptic receptors since the effect was not seen in luteal particulates. Elevation of LH receptor numbers by progesterone in immature CL may be a form of intraluteal regulation contributing to the functional maturation of these steroidogenic cells.

Immunocytochemical detection of prolactin or prolactin-like immunoreactivity in epididymis of mature male mouse

Prolactin (PRL) binds to the testis of mice and rats where it increases the number of luteinizing hormone receptors, increases the binding of human chorionic gonadotropin (hCG) to LH receptors, and enhances testosterone synthesis and secretion. PRL also binds to the prostate and seminal vesicles of rats and humans where it increases organ weight and stimulates growth and uptake of testosterone. PRL binds to the epididymis of rats but the effect of PRL on this organ is unknown. In the present study, a standard immunoperoxidase (PAP) technique was used to detect the binding of endogenous and exogenous PRL or PRL-like peptides to the epididymis of the mature mouse. Throughout the epididymal duct, a positive reaction for peroxidase, suggesting PRL or PRL-like binding, occurred in the Golgi area of principal cells. In segment 1, positive reactions were also visualized in the perinuclear area and in the region located between the Golgi area and the apical surface of the principal cells (supra-Golgi area). In the corpus and cauda epididymidis, scattered entire principal cells were also positive. Throughout the epididymal duct, the reactions indicating the binding of exogenous PRL were slightly stronger than those testing for binding of endogenous peptides. The significance of such binding to the epididymis is uncertain but PRL may perform the same functions in epididymal principal cells as it does in the testis, prostate, and seminal vesicles.

Pubertal changes in the secretion of gonadotropic hormones, testicular gonadotropic receptors and testicular function in the ram

Developmental changes in pituitary content and secretory patterns of luteinizing hormone (LH), follicle-stimulating hormone (FSH) and prolactin (PRL), testicular size and steroidogenic function, testicular LH- and FSH-binding activity, and growth of the accessory sex organs were examined for 24 Dorset × Leicester × Suffolk rams (born in March) every 30 days from 30 to 150 days of age, and again at 200 days. Pituitary LH and FSH contents increased between 30 and 60 days of age and remained constant until 150 days, when contents were somewhat greater than on either 120 or 200 days. LH-pulse amplitude and frequency, and mean FSH concentration, were highest at 60 and(or) 90 days of age. Testicular growth increased dramatically between 90 and 150 days of age in association with increases in the number of LH- (100-fold) and FSH- (33-fold) binding sites in the testis and a small increase in blood testosterone concentration (1 ng/ml). During the same period, pituitary content and blood concentration of PRL increased to maximal values, epididymal, vesicular gland and bulbourethral gland weights increased 6-fold, and body weight doubled. Between 150 and 200 days of age, testosterone concentration increased considerably (8 ng/ml), as did LH-pulse frequency and the amount of LH- and FSH- binding in the testis; the reproductive organs continued to grow at a rate faster than that of the body as a whole. Testicular development of ram lambs was accompanied by increases in the secretion of all three pituitary hormones with gonadotropic properties, and in the number of LH and FSH receptors.

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.

Rapid Leydig cell proliferation and luteinizing hormone receptor replenishment in the neonatal rat testis after a single injection of human chorionic gonadotropin.

Two-day-old rats were stimulated with a single dose of human chorionic gonadotropin (hCG). Changes in the Leydig cell number, mitotic activity, cell size, and number of luteinizing hormone (LH) receptors were studied. The Leydig cell number of the hCG-treated animals was 1.8 times that of the control on Day 1 and remained elevated for the rest of the 5-day experiment (p less than 0.0001). On Day 1 the number of Leydig cell mitoses in the hCG group was greater (p less than 0.05) than in the controls. The Leydig cell size increased transiently to two times that of the control (p less than 0.01) within the first day after the treatment and returned to control size by Day 5. The number of LH receptors per testis decreased 81% in 1 day (p less than 0.01), but returned to control level by Day 3. Since Leydig cell numbers were constant after Day 1, the rapid receptor recovery was obviously due to restoration of the binding sites rather than increased cell number. The present results demonstrate a rapid proliferative response and rapid LH receptor replenishment in the fetal-neonatal Leydig cells after gonadotropic stimulation. These responses of fetal-type Leydig cells are in clear contrast to those observed in adult testes after a similar stimulation.

Discordance in the effects of interleukin-1 on rat granulosa cell differentiation induced by follicle-stimulating hormone or activators of adenylate cyclase.

Recombinant human interleukin-1 (IL-1) inhibits the follicle-stimulating hormone (FSH)-induced development of luteinizing hormone (LH) receptors and suppresses progesterone secretion in cultured rat granulosa cells. Since activation of adenylate cyclase by FSH is considered to be the primary second messenger system responsible for differentiation of granulosa cells, we examined whether IL-1 could alter the FSH, cholera toxin, or forskolin-induced accumulation of cyclic adenosine 3', 5'-monophosphate (cAMP) from these cells. In addition, we sought to determine if IL-1 could influence differentiation induced by the cAMP analog, 8-bromo cAMP. Cells collected from ovaries of immature, diethylstilbestrol-treated rats were stimulated to differentiate by addition of FSH, cholera toxin, forskolin, or 8-bromo cAMP to the cultures. IL-1 or interleukin-2 (IL-2) was added to some of the tubes, and the primary cultures were incubated for various periods of time. At the end of the culture, the tubes were centrifuged, the medium was saved for progesterone and cAMP radioimmunoassay, and the cells were assayed for specific 125I-human chorionic gonadotropin (hCG) binding to determine the number of LH receptors. In the presence of FSH, IL-1, at a dose as small as 5 ng/ml, but not IL-2, significantly inhibited LH receptor formation and suppressed progesterone secretion in a dose-related manner. IL-1 also significantly suppressed FSH-induced cAMP accumulation after 72 h of incubation but did not appear to do so in a dose-related fashion. In the presence of FSH, IL-1 did not significantly alter the protein content of granulosa cells at the end of culture. During stimulation of granulosa cells with cholera toxin, forskolin, or 8-bromo cAMP, IL-1 significantly reduced LH receptor formation compared to that observed in the absence of IL-1. However, in contrast to IL-1 in the presence of FSH, IL-1 significantly augmented the forskolin-induced secretion of progesterone and accumulation of cAMP after 72 h at subsaturating doses of forskolin. Thus, IL-1 appeared to inhibit forskolin-induced and cholera toxin-induced formation of LH receptors even when cAMP levels were elevated. Similar to forskolin, 8-bromo cAMP-stimulated progesterone secretion was significantly enhanced by IL-1, but LH receptor formation was inhibited. Over a 72-h time course at single doses of FSH or forskolin, IL-1 did not affect cAMP accumulation until 48 h of culture, at which time IL-1 significantly suppressed FSH-induced, but augmented forskolin-induced, accumulation of cAMP.(ABSTRACT TRUNCATED AT 400 WORDS)

The corpus luteum.

Normal corpus luteum function is determined by function in the follicular as well as the luteal phase. In the follicular phase adequate follicle-stimulating hormone (FSH) and oestrogen stimulation are required for granulosa cell mitosis and luteinizing hormone (LH) receptor synthesis. An increase in LH pulse frequency may also be necessary for adequate oestrogen synthesis and preparation of follicular cells for luteinization and secretion of progesterone. The nature of LH release may also influence luteal function and pre-ovulatory progesterone may increase the responsiveness of the follicle to gonadotrophins. The thecal vascular network becomes extensive around pre-ovulatory follicles and may influence access of gonadotrophins and/or the ability of follicular cells to respond to them. Further vascularization is an early feature of luteinization. Angiogenic factors are found in luteal tissue and prostacyclin increases luteal blood flow. The corpus luteum consists of large cells which secrete most of the progesterone and have prostaglandin F2 alpha receptors and small cells which are responsive to LH. In the luteal phase subnormal luteal function has not been associated with a reduction in LH concentration, pulse frequency or amplitude. The number and occupancy of LH receptors and adenylate cyclase activity do not appear to be altered by a reduction in luteal function. Low density lipoprotein provides the substrate and somatomedin C modulates among other hormones' influences, progesterone production. In addition to the cAMP second messenger system phosphatidyl inositol metabolism may also be associated with LH stimulation. Luteolysis is an active process; prostaglandin F2 alpha or lipoxygenase products and possibly an endogenous GnRH-like ovarian hormone may mediate it as also may oxytocin in some species.

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.

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)

Luteinizing Hormone Receptors and Progesterone Content in Porcine Corpora Lutea After Prostaglandin F2α 1

The effect of prostaglandin F2 alpha (PGF2 alpha) on luteinizing hormone (LH) receptors, weight and progesterone content of corpora lutea (CL), and serum progesterone concentrations was studied in gilts. Fifteen gilts were hysterectomized between Days 9 to 11 of the estrous cycle. Twelve gilts were injected i.m. with 10 mg of PGF2 alpha and 3 with saline on Day 20. Ovaries were surgically removed from each of 3 gilts at 4, 8, 12 and 24 h following PGF2 alpha treatment and from the 3 control gilts 12 h following saline injection. Jugular blood samples for progesterone analysis were collected from all gilts at 0, 2 and 4 h following treatment and at 8, 12 and 24 h for gilts from which ovaries were removed at 8, 12 and 24 h, respectively. Mean serum progesterone and CL progesterone concentrations decreased within 4 h after PGF2 alpha treatment (P less than 0.05) and remained low through 24 h after treatment. The number of unoccupied LH receptors decreased by 4 h (P less than 0.05) and this trend continued through 24 h. There were no differences in luteal weight or affinity of unoccupied LH receptors of luteal tissue at 4, 8 12 and 24 h after PGF2 alpha when compared to luteal tissue from controls. These data indicate that during PGF2 alpha-induced luteolysis in the pig, luteal progesterone, serum progesterone concentrations and the number of LH receptors decrease simultaneously.

A central role for cyclic AMP, but not progesterone, in luteinizing hormone receptor down-regulation in the granulosa cell.

Although luteinizing hormone (LH) is known to down-regulate its own receptor in several gonadal cell types, the mechanisms underlying this process are poorly understood. To elucidate these mechanisms we have examined the role of cAMP and progesterone in LH-stimulated down-regulation of the LH receptor, using cultured granulosa cells as a model. LH receptors were induced by culturing the cells with follicle-stimulating hormone for 2 days, and once induced, could be down-regulated by a brief exposure to LH. Down-regulation also occurred when cells were cultured with activators of adenylate cyclase, inhibitors of phosphodiesterase, or analogues of cAMP. Cholera toxin and N6,O2'-dibutyryladenosine 3':5'-cyclic monophosphate, like LH, decreased the number of LH receptors, without affecting affinity for 125I-human chorionic gonadotropin (hCG). The extent of receptor loss after treatment with LH plus cholera toxin was no greater than that caused by LH alone. LH, hCG, and deglycosylated hCG, which binds to the LH receptor but has little bioactivity, caused down-regulation, and their relative capacity to cause down-regulation was highly correlated with their relative capacity to stimulate cAMP production. Indirect evidence suggested that maximal down-regulation requires activation of adenylate cyclase for at least 3 h. Consistent with this idea, a 3-h exposure to dibutyryl cAMP caused near-maximal down-regulation. Progesterone secretion was enhanced by all agents that caused down-regulation of the LH receptor; however, there was little correlation between progesterone secretion and down-regulation. Furthermore, maximal down-regulation occurred when progesterone secretion was inhibited greater than 99% with cyanoketone. These data indicate that cAMP, but not progesterone, plays a central role in LH receptor down-regulation in the granulosa cell and that elevation of intracellular cAMP levels for 3 h is both necessary and sufficient to trigger maximal down-regulation.

Luteal luteinizing hormone receptors during the postovulatory period in the mare.

Changes in serum luteinizing hormone (LH) and progesterone concentrations, number of luteal unoccupied LH receptors, receptor affinity constants, luteal weights and luteal progesterone concentrations were determined during the postovulatory period in the mare. The number of unoccupied LH receptors and receptor affinity was less during the early (Days 1-4) and late [Day 15 through 3rd day after start of corpus luteum (CL) regression] luteal phases than during the mid-luteal (Days 9-14) phase of the postovulatory period (P less than 0.01). The number of LH receptors per CL increased 21-fold (P less than 0.001) from Day 1 to Day 14. Receptor affinity increased 5-fold (P less than 0.001) from Day 1 to Day 13. Receptor number was highly correlated with receptor affinity (P less than 0.01) and both were highly correlated with serum and luteal progesterone (P less than 0.01). During regression of the CL, the number of LH receptors and receptor affinity decreased concomitantly with serum and luteal progesterone. Morphologically, luteal cell development and degeneration correlated with the change in receptor numbers, affinity constants and luteal and serum progesterone concentrations. Receptor number and affinity, luteal weight and serum and luteal progesterone concentrations did not differ between the CL from multiple ovulations. Random variations in the data observed between CL from multiple and single ovulations suggested that CL from the two groups were not different in structure and function. In summary, the above results suggest that major factors in regulation of progesterone secretion and maintenance of the equine CL are changes in the number of LH receptors and the affinity constants throughout the postovulatory period.

Developmental pattern and sensitivity to down-regulation of testicular LH receptors in the pig.

Testicular receptors for luteinizing hormone (LH) were quantified in 40 pigs between the day of birth and 22 weeks of age. The highest specific binding capacity (fmol hLH bound/mg protein) occurred at about 2--3 weeks of age which coincides with the age when the relative volume of Leydig cells is at a maximum. Significantly lower binding capacity was observed at 7 weeks of age which is the time of maximum Leydig cell regression. hCG was injected intramuscularly to immature (3000 IU, 22 days old) and adult boars (10 000 IU, 30--40 weeks old). Twentyfour h after injection the numbers of available LH receptors were reduced to 6 and 18% of control values, respectively. At this time occupied receptors were at a maximum (90% and 67% of controls, respectively). The total number of LH receptors (available + occupied) was increased 1 h after injection after which a gradual decrease was observed. Three days after hCG administration the total receptor quantities were at a minimum in both immature and adult pigs (59% and 31% of controls, respectively) and returned to control levels 5--7 days after treatment. Thus, pig LH receptors appear to be subjected both 'positive' and 'negative' regulation as previously described in the rat.

Age-related changes in the hypothalamic-pituitary-testicular function of the rat

The activity of the hypothalamic-pituitary-testicular axis was investigated in 3–4 months (young) and 24 months (old) rats. The results clearly demonstrate that aging reduces ( p < 0.01) the hypothalamic content of gonadotrophin releasing hormone (GnRH), decreases the capacity of the pituitary ( p < 0.01) to synthesize and or release follicle stimulating hormone and luteinizing hormone (LH) following a single stimulation of GnRH (50 ng/100 g body weight), lowers the capacity of the testes to produce testosterone ( p < 0.01) following multiple subcutaneous injections of human chorionic gonadotrophin (hCG 31U/100 g body weight for 3 consecutive days), decreases the number of Leydig cell LH receptors and decreases the in vitro responsiveness of the hCG-challenged Leydig cell to synthesize testosterone ( p < 0.01). These phenomena are independent of a major alteration in the capacity of the hCG challenged Leydig cell to produce adenosine 3′, 5′-monophosphate. It is concluded that the decline in testicular activity accompanying senescence is not inherent to the testes only but is also associated with alteration in the function of the hypothalamus and pituitary which eventually lead to the loss of fecundity.