Luteinizing Hormone Subunit Research Articles

Transient dwarfism and hypogonadism in mice lacking Otx1 reveal prepubescent stage-specific control of pituitary levels of GH, FSH and LH.

Genetic and molecular approaches have enabled the identification of regulatory genes critically involved in determining cell types in the pituitary gland and/or in the hypothalamus. Here we report that Otx1, a homeobox-containing gene of the Otx gene family, is postnatally transcribed and translated in the pituitary gland. Cell culture experiments indicate that Otx1 may activate transcription of the growth hormone (GH), follicle-stimulating hormone (betaFSH), luteinizing hormone (betaLH) and alpha-glycoprotein subunit (alphaGSU) genes. Analysis of Otx1 null mice indicates that, at the prepubescent stage, they exhibit transient dwarfism and hypogonadism due to low levels of pituitary GH, FSH and LH hormones which, in turn, dramatically affect downstream molecular and organ targets. Nevertheless, Otx1-/- mice gradually recover from most of these abnormalities, showing normal levels of pituitary hormones with restored growth and gonadal function at 4 months of age. Expression patterns of related hypothalamic and pituitary cell type restricted genes, growth hormone releasing hormone (GRH), gonadotropin releasing hormone (GnRH) and their pituitary receptors (GRHR and GnRHR) suggest that, in Otx1-/- mice, hypothalamic and pituitary cells of the somatotropic and gonadotropic lineages appear unaltered and that the ability to synthesize GH, FSH and LH, rather than the number of cells producing these hormones, is affected. Our data indicate that Otx1 is a new pituitary transcription factor involved at the prepubescent stage in the control of GH, FSH and LH hormone levels and suggest that a complex regulatory mechanism might exist to control the physiological need for pituitary hormones at specific postnatal stages.

ELISA for the measurement of serum and urinary chorionic gonadotropin concentrations in the laboratory macaque.

A rapid, sensitive, enzyme-linked immunosorbant assay (ELISA) for the measurement of chorionic gonadotropin (CG) in serum and urine samples of laboratory macaques is reported. The ligand (CG) is captured by a readily available, widely used, and well-characterized monoclonal antibody (Mab, 518B7) generated against the beta subunit of bovine luteinizing hormone (LH). This Mab, while specific for LH, shows very little species specificity, and has been shown to detect LH and CG by radioimmunoassay (RIA) in both human and non-human primates. A polyclonal antiserum raised in rabbits against human chorionic gonadotropin (hCG) is conjugated to horseradish peroxidase, and is used as the second antibody signal. This anti-hCG antiserum cross reacts with CG secreted by both the human (hCG) and macaque (mCG). The ELISA utilizes hCG as the standard, and results are based on the relative concentrations of mCG in serum and urine. Total assay time is less than 5 hours. Range of the standard curve is 0.002 to 0.5 ng hCG/well, and the least detectable concentration of hCG is 0.0023 +/- 0.0007 ng/well. Pregnancy was detected in early pregnant macaques (M. fascicularis) on 9 (N = 1/16), 10 (N = 1/16), 11 (N = 1/16), 12 (N = 6/16), 13 (N = 1/16), 14 (N = 4/16), and 15 (N = 2/16) days following the pre-ovulatory urinary estrone conjugate peak. The detection of pregnancy by urinary mCG occurred approximately 24 to 72 hours after its detection in serum.

Mutations of Arg68 of the human chorionic gonadotrophin beta subunit lead to reduced secretion.

The Arg68-Leu69 sequence is invariant in the beta subunits of chorionic gonadotrophin and luteinizing hormone from a variety of species. Using site-directed mutagenesis of the human chorionic gonadotrophin (hCG)-beta cDNA, several replacements of Arg68, an Ala replacement of Leu69, and a multiple replacement with Ala-Ala-Ala-Ala of the tetrapeptide sequence, Arg68-Leu69-Pro70-Gly71, were prepared and characterized. The wild-type and mutant cDNAs were subcloned into a pRSV expression vector and transiently transfected into CHO cells containing a stably integrated gene for bovine a. Concentrations of secreted wild-type and mutant hCG-beta subunit and holoprotein were determined using radioimmunoassays; potencies, i.e. the ratio of biologic to immunologic activity, of several of the mutant heterodimers were measured in vitro via gonadotrophin-mediated steroidogenesis in transformed murine Leydig cells (MA-10). The Leu69-->Ala mutant formed a mutant holoprotein that was essentially equipotent with wild-type hormone in the steroidogenesis assay. The Arg68 replacements with Lys, Ala, and Leu were poorly secreted by the cells, e.g. < 10% that of wild-type hCG; however, sufficient quantities of mutant holoproteins containing Lys68 and Ala68 were obtained for biological assays, and both exhibited greater apparent potencies than wild-type hormone. Likewise, a mutant holoprotein containing the Arg68-Leu69-Pro70-Gly71-->Ala-Ala-Ala-Ala multiple replacement was apparently more potent than wild-type hormone, but it too was secreted at lower levels than wild-type. These results establish that replacements of Arg68 in hCG-beta diminish secretion, but the small amount of holoprotein that is formed and secreted appears to be of somewhat greater potency than wild-type hormone.

Direct actions of the luteinizing hormone-releasing hormone agonist, deslorelin, on anterior pituitary contents of luteinizing hormone (LH) and follicle-stimulating hormone (FSH), LH and FSH subunit messenger ribonucleic acid, and plasma concentrations of LH and FSH in castrated male cattle.

The objective in this study was to characterize direct effects of the LHRH agonist, deslorelin, on anterior pituitary gland function in male cattle in the absence of gonadal feedback. Castrated bulls (steers), 30 mo old, were allocated to four groups: group 1, control, no treatment (n = 8); group 2, five deslorelin implants (approximately 250 micrograms total deslorelin/day) for 42 days (n = 8); group 3, control+ LHRH (50 micrograms i.m.) at weekly intervals (n = 3); group 4, five deslorelin implants+LHRH as for group 3 (n = 3). Plasma LH was similar (p > 0.05) for steers in groups 1 and 2 on Day 0 and lower (p < 0.05) for steers in group 2 on Day 4, and continued to decrease to Day 41 (group 1, 1.71 +/- 0.20 ng/ml [mean +/- SEM]; group 2, 0.38 +/- 0.03 ng/ml [p < 0.001]). Mean plasma concentrations of FSH were similar (p > 0.05) for steers in groups 1 and 2 on Day 0 and lower (p < 0.05) for steers in group 2 on Day 7, and declined to Day 41 (group 1, 43.5 +/- 3.9 ng/ml; group 2, 17.5 +/- 1.5 ng/ml [p < 0.001]). Steers in group 3 showed increases in plasma LH after injection of LHRH on all occasions, while steers in group 4 did not show increases in plasma LH from Day 14 onward. Mean relative pituitary contents (arbitrary units) of LH beta- and FSH beta-subunit mRNAs were reduced on Day 42 in steers treated with deslorelin (LH beta: groups 1 and 3, 1.56 +/- 0.27; groups 2 and 4, 0.08 +/- 0.01 [p < 0.001]; FSH beta: groups 1 and 3, 1.01 +/- 0.08; groups 2 and 4, 0.34 +/- 0.07 [p < 0.001]). However, alpha-subunit mRNA was similar for control steers and steers treated with deslorelin (groups 1 and 3, 1.00 +/- 0.11; groups 2 and 4, 0.86 +/- 0.12 [p > 0.1]). Pituitary content of LH, but not FSH, was reduced in steers treated with deslorelin. In summary, steers treated with deslorelin showed desensitization to natural LHRH, and this was associated with reduced pituitary contents of LH and FSH beta-subunit mRNAs, a reduction in pituitary content of LH, and decreases in plasma concentrations of LH and FSH. This demonstrated, for the first time, a direct action of LHRH agonist on LH and FSH beta-subunit gene expression in cattle, independent of gonadal feedback. Also, there was a differential effect of treatment with deslorelin on gonadotropin alpha- and beta-subunit mRNA contents in the anterior pituitary.

A mechanism for the differential regulation of gonadotropin subunit gene expression by gonadotropin-releasing hormone.

The hypothalamic hormone gonadotropin-releasing hormone (GnRH) is released in a pulsatile fashion, with its frequency varying throughout the reproductive cycle. Varying pulse frequencies and amplitudes differentially regulate the biosynthesis and secretion of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) by pituitary gonadotropes. The mechanism by which this occurs remains a major question in reproductive physiology. Previous studies have been limited by lack of available cell lines that express the LH and FSH subunit genes and respond to GnRH. We have overcome this limitation by transfecting the rat pituitary GH3 cell line with rat GnRH receptor (GnRHR) cDNA driven by a heterologous promoter. These cells, when cotransfected with regulatory regions of the common alpha, LH beta, or FSH beta subunit gene fused to a luciferase reporter gene, respond to GnRH with an increase in luciferase activity. Using this model, we demonstrate that different cell surface densities of the GnRHR result in the differential regulation of LH and FSH subunit gene expression by GnRH. This suggests that the differential regulation of gonadotropin subunit gene expression by GnRH observed in vivo in rats may, in turn, be mediated by varying gonadotrope cell surface GnRHR concentrations. This provides a physiologic mechanism by which a single ligand can act through a single receptor to regulate differentially the production of two hormones in the same cell.

Novel expression of luteinizing hormone subunit genes in the rat testis

The two gonadotropins, LH and FSH, are thought to be synthesized and secreted solely by the anterior pituitary. We present here evidence for expression of the LH beta and common alpha-subunit (C alpha) genes in the rat testis. The LH beta and C alpha-subunit messenger RNAs (mRNAs) were detected by reverse transcriptase-polymerase chain reaction in the rat testis and pituitary with primer pairs producing 247- and 199-base pair complementary DNA (cDNA) fragments, corresponding to nucleotides 154-400 of LH beta and nucleotides 250-448 of C alpha cDNA, respectively. The specificity of the cDNA species generated was verified by Southern hybridization using nested [32P]cDNA or oligonucleotide probes, and identity with the published rat LH beta and C alpha-subunit gene structures was determined by sequencing. The mRNA bands with specific hybridization to complementary RNA (cRNA) probes corresponding to nucleotides 154-368 of the rat LH beta cDNA and nucleotides 250-448 of the rat C alpha cDNA were found in the rat pituitary and testis by Northern hybridization. The major C alpha mRNA had a size of 0.8 kilobases (kb) in the pituitary and testis. The major LH beta transcripts were 0.8 and 2.7 kb in the pituitary and testis, respectively. To further characterize the larger testicular LH beta-subunit transcript, rapid amplification of the 3'-end of cDNA (3'-RACE) was performed using an oligo(deoxythymidine-17) adapter and a specific 5'-primer. Southern hybridization of the 3'-RACE product of rat testicular RNA with a LH beta [32P]cDNA probe had the same size as the 3'-RACE product of pituitary RNA. The pituitary and testicular RNAs were then cut into two segments using oligonucleotide-directed ribonuclease H digestion and subjected to Northern hybridization using a cRNA probe specific to the 5'-end segment. The digested 5'-end segments of the pituitary and testicular mRNAs were 0.4 and 2.3 kb, respectively, indicating that the testicular LH beta mRNA has a 1.9-kb 5'-extension, compared to the cognate pituitary mRNA. This was further verified by Northern hybridization using a cRNA probe corresponding to nucleotides -790 to -10 upstream of the pituitary initiation site of LH beta gene transcription. Specific hybridization of a 2.7-kb mRNA transcript was found in the rat testis, but none in the pituitary. Hence, the 3'-end polyadenalytion site of the LH beta mRNA is the same in rat pituitary and testis, and the different transcript sizes are due to a difference at the 5'-end.

PCR amplification and physical localization of the genes for pig FSHB and LHB

Follicle-stimulating hormone (FSH) and luteinizing hormone (LH) are members of the glycoprotein hormone family and play essential roles in gametogenesis and sexual development of mammals. Fragments of the porcine genes coding for the beta subunits of FSH and LH were amplified by PCR and used as probes for radioactive in situ hybridization in order to map these loci in the pig. Primers were chosen on the genomic DNA nucleotide sequences of FSHB and LHB as published in GenBank. Fragments of 1,127 bp (FSHB) and 1,239 bp (LHB) were cloned and verified by sequencing. FSHB was localized to pig chromosome bands 2p1.6-->p1.2 and LHB to pig chromosome band 6q2.1. The localization of LHB to the so-called halothane region of chromosome 6 could be expected from comparative mapping data. For FSHB, no conclusions can be drawn in this respect since, up to the present, too few genes are located on porcine chromosome 2.

Endocrinology: Clinical significance of invisible or partially visible luteinizing hormone

It is well known that luteinizing hormone (LH), like many other glycoproteins, is heterogeneous and presents several circulating isoforms. Recently, new sensitive immunometric assays measuring intact LH were developed. These assays have been found to underestimate or to be incapable of recognizing LH in some patients. This study was undertaken to determine the prevalence of such cases and to define their clinical characteristics. We compared three LH assays using as capture antibodies either a monoclonal antibody that reacts exclusively with intact LH (ES 600 Boehringer, Stratus Baxter) or a monoclonal antibody against the beta subunit of LH (IMX Abbott). In 17% of 90 patients tested, ES 600 measured > 50% lower LH concentrations when compared with the IMX. Moreover, in two cases LH was not detectable by ES 600 or Stratus, whereas it was normal with the IMX. We found another five such cases and discuss here the clinical data and results of different hormone measurements in these seven cases of 'invisible LH'. Although bioactive LH (mouse Leydig cell assay) was normal, the existence of low or even undetectable LH was clinically confusing and led to expensive complementary investigations such as gonadotrophin-releasing hormone analogue tests and magnetic resonance imaging. The uses and limitations of these assays are illustrated by different clinical situations in which the results of the different assays have been misleading.

A vaccine that prevents pregnancy in women.

We report here results of clinical trials on a birth control vaccine, consisting of a heterospecies dimer of the beta subunit of human chorionic gonadotropin (hCG) associated noncovalently with the alpha subunit of ovine luteinizing hormone and conjugated to tetanus and diphtheria toxoids as carriers, that induces antibodies of high avidity (K(a) approximately 10(10) M-1) against hCG. Fertile women exposed to conception over 1224 cycles recorded only one pregnancy at antibody titers of > 50 ng/ml (hCG bioneutralization capacity). The antibody response declines with time; fertility was regained when titers fell to < 35 ng/ml. This study presents evidence of the feasibility of a vaccine for control of human fertility.

Transcriptional and translational regulation of LH, prolactin and their testicular receptors by hCG and bromocriptine treatments in adult and neonatal rats

Effects of altered gonadotropin and prolactin (PRL) secretion on luteinizing hormone (LH), PRL and their testicular receptors (R) were studied in neonatal and adult rats. Changes in gene expression were monitored by measurments of steady-state mRNA levels. Five-day and 90-day-old male rats received a single s.c. injection of hCG (600 IU/kg), 1 mg/kg bromocriptine (BR) twice daily, or their combination. After 2 or 8 days, the responses of LH, PRL, their testicular R, and testosterone (T) were assessed, including measurements of the appropriate mRNA levels. Vehicle-treated age-matched animals served as controls. hCG suppressed serum LH in 2 days in adult rats from 0.85 ± 0.16 to 0.04 ± 0.01 gmg/l, and in neonates from 0.59 ± 0.29 to levels below 0.01 μg/l (p < 0.01 for both). This was accompanied at both ages by a 60% decrease in pituitary content of the LH β-subunit mRNA (p < 0.01), but a decrease in the α-chain (40%,p < 0.05) occurred only in neonates. hCG increased serum PRL in adult rats in 8 days over 2-fold (p < 0.01); this did not occur in neonates. In neonates, BR increased the LH subunit mRNAs 2-fold in 8 days (p < 0.01) without a concomitant effect on serum LH; no BR effects on the LH parameters were seen in adult animals. BR decreased pituitary PRL protein and mRNA levels at both ages (p < 0.01–0.05), but serum PRL decreased only in the adults. The homologous down-regulation of testicular LHR (near 100%) was accompanied in adults by a 30% decrease in LHR mRNA (p < 0.05). Also BR at this age decreased LHR binding (75% in 8 days, p < 0.01), but in this case no change occurred in the cognate mRNA. hCG and BR slightly up-regulated in adults PRLR binding, but only the 2-day effect of BR was accompanied by a 60% increase in PRLR mRNA (p < 0.05). In neonates, both hCG and BR increased testicular LHR and PRLR mRNA levels (p < 0.01–0.05). In adult animals, both hCG and BR suppressed testicular and serum T levels after 8 days (40–70%, p < 0.01–0.05); only BR was inhibitory to T by 8 days in the neonates (p < 0.05). In conclusion, the homologous and heterologous regulatory effects of hCG and BR on LH, PRL and their testicular R levels were only partly explained by changes in steady-state levels of the respective mRNAs. In general, the autoregulatory effects on LHR and PRLR appeared to affect steady-state levels of cognate mRNAs, whereas heteroregulation predominately involved changes at the protein level. The responses of the neonatal pituitary-gonadal axis to hCG and/or BR differed greatly from those observed in the adult, indicating that the mechanisms involved in these regulatory events in adult animals are a result of gradual postnatal development.

Heterogeneity of gonadotropins and levels of uncombined luteinizing hormone subunits in pituitaries of cryptorchid rams.

Pituitaries were collected from intact rams and rams that had been rendered bilaterally cryptorchid by surgery to examine the effects of cryptorchidism on gonadotropin heterogeneity, levels of uncombined luteinizing hormone (LH) subunits, and the apparent molecular sizes of LH and follicle-stimulating hormone (FSH). Cryptorchid rams had higher pituitary contents of LH and FSH as well as reduced testicular weights. The levels of uncombined LH subunits, their apparent molecular weights, and the apparent molecular weights of intrapituitary LH were similar in control and cryptorchid rams. However, the apparent molecular weight of intrapituitary FSH was slightly larger in cryptorchid rams. Cryptorchidism altered the pattern of gonadotropin heterogeneity by shifting the distribution of LH isoforms towards basic components and shifting the distribution of FSH isoforms towards acidic components. Thus, it appears that the altered gonadal feedback mechanisms resulting from cryptorchidism modify the pattern of both LH and FSH heterogeneity by shifting the distribution of isoforms in opposite directions.

Pituitary-gonadal interactions in perinatal rats: relationships of plasma luteinizing hormone and testosterone concentrations, and pituitary levels of LH subunit mRNAs.

The functional state of the pituitary-gonadal axis was studied in rats on days 18.5-21.5 of fetal life (f) and on day 4 postpartum by measurements of plasma levels of luteinizing hormone (LH) and testosterone (T). LH was measured using an ultrasensitive immunofluorometric assay. In addition, male fetuses were castrated and exposed to the antiandrogen flutamide (FL; 100 mg/kg BW) or the Leydig cell-specific cytotoxic agent ethylene dimethane sulphonate (EDS, 50 mg/kg BW on 2 days) by injections of the drugs to the mothers. Besides LH and T, pituitary levels of LH subunit mRNAs were measured in these animals. The results allowed the following conclusions: (1) the plasma LH levels in both sexes are low (< 0.05 micrograms/l, NIH rLH RP-2) on days f18.5 and f19.5; (2) a 4- to 5-fold increase in plasma LH occurs between days f19.5 and f20.5, and a 3- to 4-fold sex difference appears (females > males); (3) the activation of fetal testicular T production before day f19.5 takes place in the face of very low plasma LH (< 0.02 micrograms/l), suggesting that some factor(s) other than LH may stimulate the testis at this age; (4) the reciprocal changes of plasma LH and T, and the experiments with castration, EDS and FL demonstrate that testicular feedback regulation of LH secretion is functional from day f19.5 onwards; (5) the parallel changes of T in male and female fetal plasma suggest that T in female fetuses comes from male littermates, and (6) the fetal pituitary-testicular axis is less sensitive to hormonal manipulations than that of the postnatal animal, possibly due to interference of maternal and placental hormones with the pituitary-testicular interactions.

Quantitative morphological analysis of the pituitary gland in protein‐calorie malnourished rats

A quantitative analysis of the pituitary gland was conducted to ascertain the effects of protein-calorie malnutrition on the morphology of the somatotrophs, gonadotrophs, thyrotrophs, and corticotrophs. Male rats were fed a low, 8% protein diet from 20 to 50 days of age, while their age-matched controls were given a diet containing 27% protein. The hypophyses were then processed for light microscopic immunocytochemical staining using antibodies to growth hormone, the beta subunits of luteinizing hormone and thyroid stimulating hormone, and adrenal corticotrophic hormone. The number of each cell type along with an evaluation of the cell, cytoplasmic, and nuclear areas was conducted using a computerized image analyzer. All of these parameters were reduced significantly in the somatotrophs as a result of the low protein diet, while in the gonadotrophs, the cell, cytoplasmic, and nuclear areas were similarly affected. Smaller cell number, cell area, and nuclear area were noted in the corticotrophs of the malnourished animals, while in the thyrotrophs, only the cell and nuclear areas were reduced. The data demonstrate that each pituitary cell type responds in a unique manner to undernutrition.

Bovine luteinizing hormone (LH) isoforms and amounts of messenger ribonucleic acid for alpha- and LH beta-subunits in pituitaries of cows immunized against LH-releasing hormone.

Ovariectomized beef cows were actively immunized against LHRH to test the hypothesis that decreased stimulation of gonadotropes would alter the distribution of LH isoforms and amounts of mRNA for subunits of LH in the pituitary. Eight long-term (3 yr) ovariectomized beef cows were randomly assigned to one of two treatments: immunization against LHRH conjugated to human serum globulin (n = 4) and nonimmunization (control, n = 4). Mean concentration of serum LH in cows immunized against LHRH (1.0 +/- .83 ng/ml) was less (p = 0.01) than in control cows (5.0 +/- 0.83 ng/ml). Amounts of alpha- (p = 0.13) and LH beta-subunit (p = 0.10) mRNA tended to be reduced in cows immunized against LHRH compared to control cows. However, weight of the anterior pituitary and concentrations of LH in this gland did not differ (p = 0.90) among cows from the two groups. Pituitary extracts were chromatofocused on pH 10.5-7.0 gradients, and concentrations of LH in eluent fractions were determined by RIA. Extracts of all pituitaries resolved into nine isoforms (designated A through H and Z beginning with the most basic form). Only isoform F (mid-alkaline elution, pH = 8.8) was influenced by treatment (p = 0.05). Cows immunized against LHRH had a greater relative amount of isoform F (42.1 +/- 1.4%) than controls (37.2 +/- 1.4%). In summary, immunization of cows against LHRH altered 1) circulating concentrations of LH, 2) amounts of mRNA for the subunits of LH, and 3) distribution of intrapituitary LH isoforms without changing the concentrations of LH in the anterior pituitary.

Hypogonadism Caused by a Single Amino Acid Substitution in the β Subunit of Luteinizing Hormone

WEISS, JEFFREY; AXELROD, LLOYD; WHITCOMB, RANDALL W.; HARRIS, PHILIP E.; CROWLEY, WILLIAM F.; JAMESON, J. LARRY Author Information

Testosterone suppresses 8-bromo-adenosine 3',5'-monophosphate and gonadotropin-releasing hormone-stimulated luteinizing hormone subunit synthesis

Testosterone suppresses 8-bromo-adenosine 3',5'-monophosphate and gonadotropin-releasing hormone-stimulated luteinizing hormone subunit synthesis

Testosterone suppresses 8-bromo-adenosine 3',5'-monophosphate and gonadotropin-releasing hormone-stimulated luteinizing hormone subunit synthesis.

The major objective of this study was to determine the effects of testosterone (T) on 8-bromo-cAMP (8-br-cAMP)- and GnRH-stimulated LH subunit polypeptide synthesis and glycosylation in cultured male anterior pituitary cells. The anterior pituitaries from 1-week castrate male rats were enzymatically dispersed and incubated for 48 h in steroid-free medium, followed by a 48-h incubation with or without 10 nM T. The cells were then incubated for 12 h in medium containing [35S]methionine ([35S]Met) and [3H]glucosamine ([3H]Gln) with or without 1 mM 8-br-cAMP or 1 nM GnRH, with or without 10 nM T. Incorporation of radiolabeled precursors into LH subunits was determined by specific immunoprecipitation of the LH dimer with subsequent analysis of the individual LH alpha- and beta-subunits by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. LH content was quantified by RIA (iLH). Both 8-br-cAMP and GnRH stimulated iLH release. T suppressed basal and GnRH-induced iLH secretion, whereas it enhanced iLH release stimulated by 8-br-cAMP. Both 8-br-cAMP and GnRH stimulated total (cell plus medium) [35S]Met and [3H]Gln incorporation into LH alpha and LH beta, and these responses were suppressed by T. Basal [35S]Met incorporation into the LH subunits was inhibited by T, whereas T had no effect on basal levels of [3H]Gln incorporation. Neither T nor GnRH altered [3H]Gln cell uptake or incorporation into total proteins, whereas 8-br-cAMP increased these responses. There were no treatment effects on [35S]Met cell uptake or incorporation into total proteins. These results suggest that 8-br-cAMP, similar to GnRH, stimulates both polypeptide synthesis and glycosylation of the LH alpha- and beta-subunits and that T suppresses these responses to 8-br-cAMP and GnRH in a similar fashion. These data indicate that cAMP is involved in mediating the actions of GnRH on LH subunit biosynthesis and that the inhibition of LH subunit polypeptide synthesis and glycosylation by T involves postreceptor events that are regulated by cAMP-dependent mechanisms.

Steady-state amounts of alpha- and luteinizing hormone (LH) beta-subunit messenger ribonucleic acids are uncoupled from pulsatility of LH secretion during sexual maturation of the heifer.

Our primary objective for this study was to determine whether steady-state amounts of alpha- and LH beta-subunit mRNAs in the anterior pituitary are altered during sexual maturation in the bovine female. A secondary objective was to determine whether 17 beta-estradiol (E2) alters amounts of LH subunit mRNAs before onset of puberty. Heifers (7 mo old) were assigned to one of three treatments: 1) ovariectomized (OVX, n = 16); 2) OVX and administered E2 (OVXE, n = 16); or 3) ovary-intact (INTACT, n = 20). Pituitaries were collected at an estimated 120 days before onset of puberty (prepuberty) or 25 days before onset of puberty (peripuberty). Six INTACT heifers were used to determine time of puberty during the experimental period, and their pituitaries were collected 40 h after administration of prostaglandin F2 alpha (postpubertal INTACT group). Relative amounts of mRNAs for LH subunits in each pituitary were determined by Northern analysis and scanning densitometry. Amounts of alpha- and LH beta-subunit mRNAs were lower in pituitaries of INTACT heifers and OVXE heifers, regardless of stage of sexual maturation, than in those of OVX heifers. Amounts of alpha-subunit mRNA were similar in OVXE and INTACT heifers regardless of stage of sexual maturation. Amounts of LH beta-subunit mRNA did not change during sexual maturation in heifers in the INTACT group. Concentrations of E2 were higher and LH beta-subunit mRNA were lower in heifers from the prepubertal OVXE group than in heifers in all other treatment groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Hypogonadism Caused by a Single Amino Acid Substitution in the β Subunit of Luteinizing Hormone

INFERTILITY occurs in approximately 10 to 20 percent of couples and can be attributed to a reproductive disorder in the man or woman with equal frequency.1 , 2 Among men, specific causes of infertility can be identified in only a minority.1 In a previously identified kindred in which several men were infertile, the proband had not undergone spontaneous puberty and had low testosterone and elevated luteinizing hormone concentrations in serum, a pattern of hormone values indicative of primary hypogonadism.3 However, testosterone secretion in response to the administration of exogenous luteinizing hormone and human chorionic gonadotropin was normal. Thus, the proband appeared to . . .

Histochemical and immunocytochemical identification of the pituitary cell types in three sciaenid fishes: Atlantic croaker ( Micropogonias undulatus), spotted seatrout ( Cynoscion nebulosus), and red drum ( Sciaenops ocellatus)

The adenohypophysial cell types in three species of sciaenid teleosts were identified using a combination of classical histochemical techniques and immunocytochemistry with antisera raised against piscine and human pituitary hormones. In general greater specificity for the different cell types was observed with antisera to the piscine pituitary hormones than with antisera raised against the human hormones. The distribution of the cell types did not differ significantly among the three species, Atlantic croaker ( Micropogonias undulatus), spotted seatrout ( Cynoscion nebulosus), and red drum ( Sciaenops ocellatus), and was similar to that reported in several other teleosts. A major portion of the rostral pars distalis was composed of acidophilic prolactin cells which showed specific binding to antisera to carp, coho salmon, and chinook salmon prolactins. The acidophilic corticotrops formed cords of cells which bordered the prolactin cells and showed strong immunoreactivity with anti-human ACTH. This antiserum also cross-reacted with the lead hematoxylin-positive cells in the pars intermedia. The acidophilic growth hormone cells were restricted to the dorsal part of the proximal pars distalis in close contact with the neurohypophysis and were specifically bound by anti-carp and anti-chum salmon growth hormone. The basophilic thyrotrops were located in the dorsal-anterior part of the proximal pars distalis and showed strong immunoreactivity to antiserum to the human thyrotropin β subunit. This antiserum also bound weakly to the basophilic gonadotrops which were concentrated in the ventral part of the proximal pars distalis and also extended posteriorly to form a border around the pars intermedia. Antiserum to croaker maturational gonadotropin, which recognizes two distinct gonadotropins in sciaenids, bound strongly and specifically to the gonadotrops. The gonadotrops were also bound specifically by antisera to the α and β subunits of human luteinizing hormone. Antisera to the β subunits of two recently isolated coho salmon gonadotropins, GTH-I and GTH-II, failed to bind the gonadotrops strongly or differentially in the three sciaenid species. Consequently, the number of gonadotropin cell types in sciaenid fishes remains unresolved.