Biomedical subjects

W Vale

Publications and source records attributed to W Vale.

At least 163 records · Page 9Linked to original sources

PubMed1989

Inhibin and activin modulate the release of gonadotropin-releasing hormone, human chorionic gonadotropin, and progesterone from cultured human placental cells.

Although it is clear that human chorionic gonadotropin (hCG) and progesterone play fundamental roles in pregnancy, the regulation of placental production of these hormones remains to be defined. Recent evidence suggests that the human placenta expresses proteins related to inhibin (alpha beta subunits) or activin (beta beta subunits). Inhibin and activin (follicle-stimulating hormone-releasing protein) possess opposing activities in several biological systems including pituitary follicle-stimulating hormone (follitropin) secretion, erythroid differentiation, and gonadal sex-steroid production. The actions of purified inhibin and activin on hormonogenesis by primary cultures of human placental cells were studied. The addition of activin increased gonadotropin-releasing hormone (GnRH) and progesterone production and potentiated the GnRH-induced release of hCG. Inhibin by itself did not modify placental immunoreactive GnRH, hCG, and progesterone secretion but reversed the activin-induced changes. Neither inhibin nor activin influenced the release of human placental lactogen. Furthermore, transforming growth factor beta, structurally related to inhibin/activin, did not significantly influence hormone release from cultured placental cells. These results support the hypothesis that inhibin and activin may play a role in regulating the release of GnRH, hCG, and progesterone from placenta and implicate inhibin-related proteins in the endocrine physiology of human pregnancy.

Activins↗

PubMed1989

Possible involvement of inhibin in altered follicle-stimulating hormone (FSH) secretion during dissociated luteinizing hormone (LH) and FSH release: unilateral castration and experimental cryptorchidism.

Male rats were either unilaterally or bilaterally castrated, or were rendered cryptorchid when they were either 15 or 45 days old. Subsequently, blood was sampled over the next several weeks and plasma luteinizing hormone (LH), follicle-stimulating hormone (FSH), testosterone (T), and immunoreactive inhibin-alpha (irI alpha) levels were measured by specific radioimmunoassays (RIAs). At the end of the experiment, gonadal expression of inhibin-alpha, inhibin-beta A, and inhibin-beta B subunits was measured by S1 nuclease analysis and in situ hybridization. In both age groups, bilateral castration (BC) produced the expected marked (p less than or equal to 0.01) increases in plasma LH and FSH levels, and concomitant decreases in T and irI alpha secretion within 1 - 2 days after surgery. In 15-day-old animals, unilateral castration (UC) significantly increased FSH and decreased circulating levels of irI alpha, but did not measurably alter LH or androgen production. At 7 days after surgery, the level of inhibin mRNA in the remaining testis was unchanged. In 45-day-old animals, UC caused a measurable increase in FSH, with little or no changes in the circulating levels of irI alpha. Plasma T levels were lowered (p less than or equal to 0.05) by UC; however, there were no statistical changes in LH levels in these UC rats. Finally, T administration markedly reversed UC-induced increase in FSH secretion in both age groups. Androgen therapy also interfered with inhibin release in 45-day-old, but not in 15-day-old rats. In rats 15 days old at the time of surgery, cryptorchidism produced a small but measurable increase (p less than or equal to 0.05) in LH release at Week 6 only, which was accompanied by a significant (p less than or equal to 0.01) decline in T secretion. Plasma FSH levels were elevated at all times in cryptorchid rats, and at 2, 4, and 6 wk, these levels were not statistically distinguishable (p greater than 0.05) from those of castrated animals. In this group of rats, cryptorchidism caused a transient increase (p less than or equal to 0.05) in irI alpha values 1 wk after surgery, but no changes at later times. Finally, measurement of testicular inhibin-alpha subunit messenger RNA (mRNA) levels showed an approximately 2-fold increase compared to total RNA levels in the testis. However, because of the significant decrease in total RNA levels per testis caused by cryptorchidism, the absolute change in inhibin-alpha subunit mRNA levels per testis corresponded to an approximately 3-fold decrease.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1989

Immunoreactive inhibin secretion by the hypophysectomized female rat: demonstration of the modulating effect of gonadotropin-releasing hormone and estrogen through a direct ovarian site of action.

To investigate the possibility that sex steroids and GnRH might act directly at the level of the ovary to modulate the secretion of immunoreactive inhibin, we have administered estradiol benzoate, PMSG, or GnRH analogs to immature (24-day-old) hypophysectomized female rats. The sc injection of 10-100 micrograms estradiol benzoate markedly (P less than or equal to 0.01) increased plasma inhibin levels measured 48 h later. Administration of the GnRH agonist [DTrp6,Pro9-NEt]GnRH significantly (P less than or equal to 0.05) inhibited both spontaneous and PMSG-induced inhibin secretion, while the GnRH antagonist [Ac-D2Nal1,D4ClPhe2,D3Pal3,Arg5,DGlu6(AA), DAla10]GnRH augmented the stimulatory effect of PMSG action on inhibin release. These results suggest a direct ovarian site of action of sex steroids and GnRH analogs in modulating the release of immunoreactive inhibin. They also support the hypothesis that an endogenous GnRH-like peptide of ovarian origin plays a physiological paracrine role in modulating inhibin secretion in the rat.

Animals↗

PubMed1989

Nucleotide regulation of growth hormone-releasing factor binding to rat pituitary receptors.

The specific binding of a GRF radioligand, [His1,125I-Tyr10,Nle27]hGRF-1-32NH2, to rat pituitary homogenates is reduced by the addition of GTP and its nonhydrolyzable analogs 5'-guanylylimidodiphosphate (GppNHp) and guanosine 5'-O-(3-thiotriphosphate) (GTP-gamma-S). GDP and cAMP had no effect while the nonhydrolyzable ATP analogs 5'-adenylylimidodiphosphate and adenosine 5'-O-(3-thiotriphosphate) did elicit a significant reduction in GRF binding. The effect of GppNHp was half-maximal at 0.2 microM, and the maximum inhibition achieved was 85%. The effect of 0.1 microM GppNHp on GRF competitive displacement experiments indicated a significant reduction in affinity for the ligand (Kd = 0.51 +/- 0.11 nM in the absence of GppNHp and 2.1 +/- 1.1 nM in its presence) without an effect on receptor number. The GRF radioligand dissociates slowly from its receptor (t1/2 = 250 +/- 50 min), but the addition of 0.1 microM GppNHp converts approximately half of the receptors present to a more rapidly dissociating form (t1/2 = 9 +/- 10 min). These results are consistent with existing models for receptor-G-protein interactions, and thus, we conclude that transduction of the GRF response across the cell membrane involves a guanine nucleotide-binding protein, presumably Gs.

Adenosine Triphosphate↗

PubMed1989

Production and regulation of inhibin subunits in pituitary gonadotropes.

Two related proteins, inhibin and activin, are produced and secreted by the gonads and act at the pituitary to regulate FSH secretion. In the present study, the alpha and beta B, but not the beta A, polypeptide subunits of inhibin were localized in the cytoplasm of FSH- and LH-immunoreactive (ir) gonadotropes. Ovariectomy (OVX) increased the size and number of cells immunoreactive for inhibin-alpha and -beta B as well as the mRNAs encoding these subunits. Treatment with estrogen prevented these effects. These results suggest that pituitary gonadotropes are sources, as well as targets, of inhibin-related peptides, whose expression in the pituitary is modulated by ovarian factors.

Animals↗

PubMed1989

Secretion of corticotropin-releasing factor from cultured rat hypothalamic cells: effects of catecholamines.

An understanding of the regulation of CRF secretion in rats is currently incomplete, in part due to the lack of sensitive in vitro models available for studying this neuropeptide. In particular, the effects of catecholamines on CRF secretion, and the receptor subtypes mediating these actions have long been the subject of much debate. A cultured cell model has been adapted for studying secretory responses of hypothalamic cells of 1-week-old rats. Between 7-16 days in monolayer culture the cells secreted detectable levels of immunoreactive CRF, and this release was paralleled by the appearance of punctate bead-like regions of immunoreactivity along fine cellular processes. CRF secretion was increased up to 4-fold by norepinephrine (EC50, approximately 0.5 microM). The increase in CRF secretion produced by norepinephrine was blocked by the beta-receptor antagonist propranolol, but not by the alpha-antagonist prazosin. Moreover, the beta-receptor agonist isoproterenol significantly elevated CRF secretion, whereas the alpha-agonist phenylephrine was without effect, except at high concentrations. Addition of phenylephrine, however, potentiated the effect of isoproterenol, but this response was still significantly less than that produced by norepinephrine. Forskolin (EC50, approximately 0.7 microM) and the active phorbol ester 12-O-tetradecanoyl-phorbol-13-acetate (EC50, approximately 40 nM) also increased CRF secretion by 3- to 4-fold. Inactive phorbol derivatives had no effect on CRF release from these cultures. The results indicate that cultured neonatal rat hypothalamic cells are a powerful model for the study of CRF release in vitro, and that norepinephrine acts directly at the isolated cell level to stimulate secretion of this peptide, primarily by activating beta-adrenoceptors. The results also suggest that at least two functional second messenger systems (adenylate cyclase and protein kinase-C) are involved in CRF secretion and are already functional in the neonatal hypothalamus.

Animals↗

PubMed1989

The gonadotropin-releasing hormone pituitary receptor interacts with a guanosine triphosphate-binding protein: differential effects of guanyl nucleotides on agonist and antagonist binding.

Binding of the GnRH agonist [DAla6,NMe-Leu7,Pro9Net]GnRH to bovine anterior pituitary membranes is inhibited by guanyl nucleotides. The effect of guanyl nucleotides is temperature dependent, in that significant binding inhibition is observed when the receptor-hormone interaction is measured at 37 C, and no inhibition is seen at 4 C. Micromolar concentrations of the nonhydrolyzable GTP analog 5'-guanylylimidodiphosphate [Gpp(NH)p] displace the bound agonist in a dose-dependent manner, with half-maximal displacement occurring in a concentration range of 0.1-0.5 microM, and maximum displacement occurring at a concentration of 50 microM Gpp(NH)p. At a concentration of 50 microM, the other nucleotides GTP and GDP inhibit binding to a lesser extent, while GMP, cGMP, 5'-adenylylimidodiphosphate [App(NH)p], ATP, and cAMP have no effect on the binding. At 37 C, Gpp(NH)p reduces the affinity of the agonist by a factor of 6 and increases its dissociation rate. In the presence of Gpp(NH)p at 37 C, there is also a 2-fold increase in the total number of binding sites. Under the same conditions as those used for the agonist, there is no displacement of the bound antagonist [Ac-D2Nal1,4ClDPhe2,D3Pal3,DLys6,Lys8,D Ala10]-GnRH by doses up to 50 microM Gpp(NH)p. The modulation of the binding of the agonist, but not that of the antagonist, by guanyl nucleotides is characteristic of receptors that are coupled to GTP-binding proteins. Thus, the GnRH receptor appears to be coupled to a GTP-binding protein that may play a role in the mechanism of action of GnRH at the pituitary.

Animals↗

PubMed1989

Oxytocin mediates the hypothalamic action of vasoactive intestinal peptide to stimulate prolactin secretion.

The ability of centrally administered vasoactive intestinal peptide (VIP) to stimulate PRL secretion when injected intracerebroventricularly could be due to leakage to the pituitary, where it is known to exert direct PRL-releasing activity, or to a hypothalamic action on its own release or that of another possible PRL-releasing factor. When 3 micrograms VIP were injected into the third ventricle of conscious ovariectomized rats, a significant (P less than 0.005) and transient elevation of plasma oxytocin (OT) levels was observed. When OVX rats were injected iv with 1 ml anti-OT serum 30 min before the central administration of 3 micrograms VIP, the PRL surge seen after VIP injection in normal rabbit serum-treated controls was completely absent. The PRL surge seen after central VIP administration was not significantly altered by iv saline infusion (1 ml over 30 min) or by infusion of a VIP antagonist [D-4-Cl-Phe6,Leu17]VIP at a dose of 0.5 microgram/kg.min in 1 ml saline for 30 min before the VIP injection. This was not due to the inability of the VIP antagonist to block the PRL-releasing factor activity of VIP, since it significantly antagonized that action both in vitro and in vivo in the suckling stimulation paradigm. However, the PRL surge was completely absent in ovariectomized rats pretreated by iv infusion of an OT antagonist, [deamino Cys1,D-Trp2,Val4,Orn8]OT, at a similar dose. This recruitment of OT by VIP indicates that it may act at more than one locus within the hypothalamo-pituitary axis to insure the coordinated control of PRL secretion.

Animals↗

PubMed1989

Identification of immunoreactive neuropeptide-gamma in human placenta: localization, secretion, and binding sites.

The aim of the present study was to evaluate the possible presence of immunoreactive neuropeptide-gamma (irNPY) in human placenta. Acidic extracts of human placental tissue collected at term pregnancy contained high irNPY concentrations. The extracted irNPY eluted from HPLC with the same retention time as synthetic NPY. The presence of the peptide in placental cells was confirmed by immunohistochemical findings showing numerous cells of the cytotrophoblast layer positively staining for NPY. Further supporting local production of the peptide, primary cultures of human placental cells released irNPY into the culture medium and the addition of high K+ concentrations increased the release of the peptide. The finding of irNPY in human placenta stimulated the characterization of binding sites of NPY in the same tissue. Using autoradiographic techniques we showed specific binding of [125I]NPY in human placental tissue. The binding of [125I]NPY to the placental receptors was saturable and widely distributed within the placental tissue. Finally, the addition of NPY to the medium of cultured placental cells increased the release of immunoreactive CRF, suggesting a possible role of NPY in placental hormone production. The effect of NPY was dose related and augmented by the addition of norepinephrine (10 nM). These results showed that human placenta produces and secretes irNPY and that NPY receptors are present in placental tissue. Moreover, the evidence that NPY stimulated the release of immunoreactive CRF from cultured placental cells suggests an action of NPY in placental hormonogenesis.

Autoradiography↗

PubMed1989

In the rat, interleukin-1 alpha acts at the level of the brain and the gonads to interfere with gonadotropin and sex steroid secretion.

These studies were designed to investigate a possible effect of the monokine interleukin-1 alpha (Il-1 alpha) on gonadotropins and/or sex steroid secretion. Three putative sites of action were considered: the central nervous system, the pituitary, and the gonads. A central nervous system-mediated mechanism was tested by injecting Il-1 alpha into the lateral ventricle of the brain of castrated rats; this treatment caused a dose-related inhibition of LH secretion, with a minimum effective dose below 0.1 microgram (6.6 pmol) and a maximum effect at 1 microgram (66 pmol). The plateau response was observed 120 min after injection. The possibility that Il-1 alpha might alter pituitary sensitivity was studied by administering 200 ng GnRH, iv, into gonadectomized control rats or rats pretreated with 1 micrograms Il-1 alpha intracerebroventricularly. GnRH induced a comparable increase in the plasma LH levels of both groups of animals. Finally, hypophysectomized immature (24-day-old) female rats were used to determine if we could confirm in vivo that interleukin-1 alpha can act directly at the level of the ovary to interfere with steroidogenesis. In these animals, the sc injection of 20 IU PMSG produced a marked (P less than 0.01) increase in plasma estradiol levels, while the sequential injection of 20 IU PMSG and 1 microgram hCG significantly (P less than 0.01) stimulated progesterone release. The ip administration of 1 microgram Il-1 alpha every 12 h to gonadotropin-treated rats resulted in a significant inhibition of both estradiol and progesterone secretion. The results support possible roles for Il-1 alpha at the levels of the brain and the gonads, but not the pituitary, to inhibit reproductive functions.

Animals↗

PubMed1989

Immunoneutralization of endogenous inhibin modifies hormone secretion and ovulation rate in the rat.

Studies were conducted to determine the effects of inhibin immunoneutralization on gonadotropins and sex steroids secretion, as well as ovulation rate and the outcome of pregnancy in the rat. These changes were compared to the normal pattern of hormone release during the estrous cycle, as well as to the ovulatory and gestational history of control animals. Measurement of plasma inhibin levels with antibodies raised against the N-terminus part of the alpha-chain of the molecule indicated an increase in immunoreactive inhibin-alpha release during the afternoon of proestrus, followed by a marked decline after the primary gonadotropin surge. Removal of endogenous inhibin by the iv injection of anti-inhibin serum on diestrus-1 caused the expected rise in plasma FSH, but not LH values. FSH secretion remained elevated for approximately 28 h, then returned to baseline. While not altering vaginal cytology, the injection of the antiserum during diestrus-1 or -2, and to a lesser degree, during estrus, caused a marked increase in the number of tubal ova shed during the subsequent estrus. However, plasma estrogen and progesterone remained comparable at all times between control and treated animals. In female rats mated after inhibin immunoneutralization at diestrus-1, the increase in ovulation rate resulted in a significant increase in developing fetuses, but only ca. 40% of the pups survived. In order to assess whether elevated FSH secretion represented the primary mechanism through which removal of endogenous inhibin increased the number of maturing follicles, ovine FSH was infused for 35 h starting in diestrus-1 or diestrus-2. Similar to the treatment with the anti-inhibin serum, FSH infusion caused an increase the number of eggs present in the oviducts on the next day of estrus. These results support the existence of a close functional relationship between the circulating levels of FSH and inhibin in the female rat. They also indicate that immunization against inhibin increases ovulation rate, a phenomenon probably mediated through elevated levels of circulating FSH.

Animals↗

PubMed1989

Possible role of luteinizing hormone and follicle-stimulating hormone in modulating inhibin secretion and expression during the estrous cycle of the rat.

In the female rat, plasma immunoreactive inhibin alpha (irl alpha) levels show marked changes during proestrus and estrus. We investigated the modulating effect of LH and FSH on these changes by injecting the GnRH antagonist DNal-DCpa-DPal-Dpr-(Ac)Dal-Leu-Arg-Pro-Asn-NH2, with or without exogenous LH replacement. Administration of the antagonist at noon on proestrus abolished the primary (proestrus) LH and FSH surge and markedly reduced the secondary (estrus) FSH surge. This treatment also reduced the release of irl alpha normally measured during proestrus afternoon, and partially prevented the decrease in irI alpha secretion on proestrus evening. Exogenous LH injected at 1545 h on proestrus had no measurable effect on irI alpha or FSH levels in control rats; however, in antagonist-treated animals, it restored the secondary FSH surge to control values while augmenting the late proestrus fall in irI alpha. This suggests that the decrease in inhibin secretion measured after exogenous LH treatment represents the mechanism through which LH induced the secondary FSH surge in antagonist-blocked rats. We also used in situ hybridization techniques to examine the changes in the expression of inhibin subunits in the ovary at 0200 h on estrus. The antagonist reduced expression of the alpha-, beta A-, and beta B-subunits in all follicle and tissue types, with the exception of the granulosa cells of large tertiary (possibly preovulatory) follicles where the signal appeared greatly enhanced. These changes were reversed by LH. The alteration in inhibin subunit messages caused by blockade of the primary gonadotropin surge suggests the presence of a cross-regulation between LH and inhibin/activin secretion, so that a decline in circulating LH levels might stimulate inhibin/activin secretion in the granulosa cells of preovulatory follicles, while reducing the production of these proteins in less mature follicles and in other ovarian cell types.

Animals↗

PubMed1989

Characterization of melanin-concentrating hormone from rat hypothalamus.

A melanin-concentrating hormone (MCH)-like peptide was isolated from rat hypothalamus by acid extraction, gel filtration chromatography, immunoaffinity chromatography using antiserum directed against salmon MCH, and two steps of HPLC using octadecyl columns. Several zones of immunoreactivity were isolated, and Edman degradation in a gas phase sequencer indicated that the amino acid sequence of all zones was identical. Rat hypothalamic MCH is a nonadecapeptide which differs from salmon MCH by an N-terminal extension of two amino acids and four additional substitutions. Rat MCH has the following primary structure: Asp-Phe-Asp-Met-Leu-Arg-Cys-Met-Leu-Gly-Arg-Val-Tyr-Arg-Pro-Cys-Trp-Gln- Val.

Amino Acid Sequence↗

PubMed1989

Activin-A, inhibin and transforming growth factor-beta modulate growth of two gonadal cell lines.

The proliferative and differentiating effects of the gonadal hormones inhibin and activin-A were examined on cell lines derived from the ovary and testis. Activin-A was found to inhibit the growth of CHO-K1 (Chinese hamster ovary) cells in culture, with an IC50 of 3 ng/ml. The maximal response (50% inhibition) required 3 days of incubation in the presence of 40 ng/ml activin-A, and the inhibitory effect was accompanied by morphological changes. Inhibin (10 ng/ml) partially blocked the inhibition of growth by activin. Transforming growth factor-beta (TGF beta), which is structurally related to activin and inhibin, was a very potent inhibitor of the proliferation of CHO-K1 cells, with an IC50 of 0.2 ng/ml and a maximal effect (70% inhibition) at 2 ng/ml. The combination of high concentrations of both TGF beta and activin-A did not result in a greater inhibitory effect than that observed with TGF beta alone, suggesting an overlapping step in the mechanism of action for both factors. In contrast to the results with CHO-K1 cells, differential effects of activin-A and TGF beta were observed in R2C (rat Leydig cell testicular tumor) cells. Activin-A had only a slight effect on proliferation over a 4-day incubation, but inhibited progesterone accumulation ina concentration-dependent fashion within 12 h. TGF beta, on the other hand, was a potent inhibitor of both growth and steroidogenesis in R2C cells. These studies suggest that activin-A and inhibin may regulate proliferation as well as functions of gonadal cells.

Activins↗

PubMed1989

Effects of transforming growth factors and inhibin-related proteins on rat preovulatory graafian follicles in vitro.

In view of recent reports on ovarian production and action of transforming growth factors (TGFs) and inhibin-related proteins (inhibin, activin, and follistatin), we have examined the effects of these hormones on the function of preovulatory follicles in vitro. Individual preovulatory follicles were obtained from PMSG-treated rats and incubated with these hormones in the absence or presence of LH. Oocyte maturation and progesterone production were monitored. Treatment with TGF alpha alone, but not with TGF beta or inhibin-related proteins, mimicked the action of LH on oocyte maturation by inducing the resumption of meiosis in follicle-enclosed oocytes (56.6% and 80.6% oocytes resumed meiosis in the presence of 0.5 and 1.0 microgram/ml TGF alpha, respectively). In follicle cultures treated with LH to induce oocyte maturation, cotreatment with inhibin and TGF beta (30-50 ng/ml), but not other related hormones, partially inhibited LH-induced meiosis in follicle-enclosed oocytes (from 82% mature ova in the presence of LH to 51% and 55% mature ova with TGF beta and inhibin, respectively). In contrast to follicle cultures, none of the hormones tested significantly affected the spontaneous maturation of rat oocytes explanted from their follicles and cultured within their cumulus mass for 4 h. Treatment with TGF alpha, but not with TGF beta, inhibin, activin, or follistatin, stimulated progesterone production. The present study demonstrated that TGF alpha, like LH, induces oocyte maturation and progesterone production in preovulatory rat follicles. Furthermore, inhibin and TGF beta suppressed LH-induced resumption of meiosis in follicle-enclosed oocytes. Because these growth factors and inhibin-related proteins are synthesized by follicle cells, they may play important roles in regulating follicular development and activity.

Animals↗

PubMed1989

Immunostainable inhibin subunits are in multiple types of testicular cells.

Immunostainable inhibin alpha-subunit has been demonstrated in rat testes in a pattern consistent with localization in Sertoli cells. In the present study the distribution of alpha-subunit immunostaining was compared to those of beta-A- and beta-B-subunits. Immunostaining of alpha-subunit was present in the seminiferous epithelium of fetal, neonatal, pubertal, and adult rats as well as in Sertoli cells in culture. The distribution of inhibin beta-B-subunit immunostaining in this epithelium was consistent with Sertoli cell localization similar to that of the alpha-subunit. The predominant staining with antibodies against the beta-A-subunit was in nuclei of immature germ cells around the periphery of each seminiferous tubule. The most probable localization of this staining was in the nuclei of pachytene and zygotene spermatocytes. Specific immunostaining with beta-A-subunit antiserum was also evident in the seminiferous epithelium adjacent to the tubular lumen. Immunoreactive alpha- and beta-A-subunit staining was present in a Leydig cell line, and beta-A immunoreactivity was present in interstitial cells of neonatal rat testes. After hypophysectomy, inhibin alpha-subunit immunostaining decreased, beta-A-subunit staining did not change, and beta-B-subunit staining increased. We conclude that immunoreactive inhibin subunits are present in multiple cells in the testis and that the amounts of immunostainable subunits in the seminiferous epithelium are differentially regulated.

Animals↗

PubMed1989

The rat melanin-concentrating hormone messenger ribonucleic acid encodes multiple putative neuropeptides coexpressed in the dorsolateral hypothalamus.

The melanin-concentrating hormone (MCH) is a cyclic neuropeptide, first isolated from salmon pituitary glands, which regulates melanin dispersion in the skin and perhaps the activity of the pituitary-adrenal axis in teleost fish. We have recently purified and characterized rat MCH (rMCH) and report here the cloning and sequencing of specific MCH cDNA isolated from a rat hypothalamic library. The sequence of rMCH found by DNA sequencing confirms the sequence deduced from the purified peptide. rMCH is located at the C-terminus of a protein precursor of 165 amino acid residues. Comparison of the amino acid sequence of prepro-MCH and that of the Aplysia peptide-A prohormone suggests that these proteins as well as other precursors may be evolutionarily related. Besides rMCH, two putative neuropeptides, termed NGE and NEI, might be generated from the same precursor. The rMCH precursor shared sequence identities with human GH-releasing factor and mammalian CRF in the regions encoding NGE and NEI. By immunohistochemical studies we have established that the amidated C-terminus of NEI is recognized by some alpha MSH and rat CRF antisera and that the C-terminal portion of NGE is responsible for the cross-reactivity revealed with one hGRF-(1-37) antiserum. Our results explain the staining of a discrete population of dorso-lateral hypothalamic neurons by heretofore seemingly unrelated antisera and provide evidence for the production of multiple novel neuropeptides from a common precursor.

Animals↗

PubMed1989

Regulation of Leydig cell function in primary culture by inhibin and activin.

Inhibin and activin are gonadal glycoproteins that selectively inhibit and stimulate FSH release, respectively. Previously we have reported that transforming growth factor-beta inhibited hCG-stimulated testosterone formation in mature Leydig cells. In the present study we evaluated the effects of other members of the transforming growth factor-beta family, inhibin and activin, on Leydig cell function. We found that activin (0.1-10 ng/ml) had no effect on basal testosterone formation, but inhibited hCG-stimulated testosterone formation in a dose-dependent manner. Activin (10 ng/ml) inhibited hCG-stimulated testosterone formation by 42%. Activin also inhibited hCG-stimulated cAMP formation. In the presence of activin (5 ng/ml), forskolin (10 microM)- and 8-bromo-cAMP (0.1 mM)-induced testosterone formation were reduced about one third. Conversions of pregnenolone and progesterone to testosterone were also blocked by activin. Interestingly, [125I]hCG binding to Leydig cells and forskolin-induced cAMP formation were not affected by the addition of activin. In contrast to activin, inhibin (0.1-10 ng/ml) had no effect on hCG-induced testosterone formation at any concentration used. However, the inhibitory effects of activin on Leydig cell function were reversed by the concomitant addition of inhibin. Our results suggest that activin inhibits testosterone formation by the Leydig cells derived from normal mature rats. Multiple steps of the steroidogenic pathway are affected by testosterone. Inhibin alone has no effect, but reverses the inhibitory action of activin.

8-Bromo Cyclic Adenosine Monophosphate↗