Plurihormonality in the secretory granules of the normal human pituitary. An immunoelectron microscopic study

Normal human autopsy anterior pituitary tissue from 5 cases was embedded in LR White resin and immunolabelled using silver-enhanced 5-nm protein A gold probes. Follicle stimulating hormone (FSH), thyroid stimulating hormone (TSH), luteinizing hormone (LH), adrenocorticotrophic hormone (ACTH), growth hormone (GH) and prolactin (PRL) were immunolocalised to the level of secretory granule. A two-sided double-labelling method was used to cross-react two hormones at a time with respect to their corresponding antibodies. All possible combinations of the six pituitary hormones were tested. Plurihormonal granules were found that contained LH + FSH, LH + TSH, and FSH + TSH. Each hormone was also found in monohormonal granules. Granule diameter was significantly larger in the pluri as opposed to monohormonal granules.     

Plurihormonality in the secretory granules of the normal human pituitary. An immunoelectron microscopic study

Summary Normal human autopsy anterior pituitary tissue from 5 cases was embedded in LR White resin and immunolabelled using silver-enhanced 5-nm protein A gold probes. Follicle stimulating hormone (FSH), thyroid stimulating hormone (TSH), luteinizing hormone (LH), adrenocorticotrophic hormone (ACTH), growth hormone (GH) and prolactin (PRL) were immunolocalised to the level of secretory granule.A two-sided double-labelling method was used to cross-react two hormones at a time with respect to their corresponding antibodies. All possible combinations of the six pituitary hormones were tested. Plurihormonal granules were found that contained LH+FSH, LH+TSH, and FSH+TSH. Each hormone was also found in monohormonal granules. Granule diameter was significantly larger in the pluri as opposed to monohormonal granules.     

Gonadotrophin-releasing activity of histones H2A and H2B

We report that histones H2A and H2B possess gonadotrophin-releasing activity in vitro and assess the signal transduction pathways involved in these effects. Perifused and incubated rat anterior pituitary (AP) cells were used, and luteinizing hormone (LH) and follicle stimulating hormone (FSH) were measured by RIA. Perifusion of cells with histone H2A (30 μM) or histone H2B (30 μM), markedly stimulated LH release but failed to elicit any FSH response. Cells incubated with 6 or 30 μM histone H2A showed a dose- and time-dependent stimulatory effect on both LH and FSH release which was blocked by 1 μM peptide MB35, an 86–120 amino acid fragment of histone H2A. Incubation of pituitary cells with gonadotrophin-releasing hormone (GnRH) and histones H2A or H2B showed a stimulatory effect on LH and FSH release which was similar to the sum of the separate effects. Trifluoperazine, as well as ethylene glycol bis(b-aminoethyl ether) N,N,N′,N′-tetraacetic acid (EGTA), alone or in the presence of the calcium ionophore A23187, significantly reduced the response of AP cells to histones. Various cyclic adenosine monophosphate (cAMP) enhancers had no effect on histone-stimulated release of gonadotrophins in incubated AP cells. Our results confirm previous evidence that histones may act as hypophysiotrophic signals. Calcium- and diacylglycerol-associated pathways, but not cAMP, appear to participate in these effects. Received 11 August 1997; received after revision 20 January 1998; accepted 26 January 1998     

Pituitary-testicular relationships in the rat embryo]

LH given to the Rat embryo at the time of hypophysectomy by decapitation (18 days of intrauterine life) will prevent the drop in androgen synthesis which normally ensues. This is regarded as definite proof that androgen synthesis is under pituitary control in the rat embryo at late gestational stages. On the other hand, the decrease in androgen synthesis following testosterone injection to the pregnant female points to the fact that there is a negative feedback of testosterone on pituitary LH secretion.     

Effects of a bovine pineal peptidic fraction (E5) on plasma and pituitary levels of LH,FSH and prolactin

Summary Pituitary levels of prolactin were significantly diminished after 5 s.c. injections spaced 12 h apart of 5 g of a partially purified bovine pineal peptide fraction (E₅) in both intact and castrated rats. E₅ treatment did not significantly affect the castration-induced changes in plasma luteinizing hormone (LH), follicle-stimulating hormone (FSH) or pituitary LH but did partially block the pituitary fall in FSH in castrated animals.Supported by NSF grant PCM 8003441 and NIH Center for Reproductive Biology (Bioassay Core) No.P30 HD 10202. Many thanks are due A. Moreno for his excellent technical assistance.     

Hormonal control of mammalian oocyte meiosis at diplotene stage

Mammalian oocytes grow and undergo meiosis within ovarian follicles. Fully grown oocytes are arrested at the first meiotic prophase by a mural granulosa origin “arrester” until a surge of luteinizing hormone (LH) from the pituitary at the mid-cycle stimulates the immature oocyte to resume meiosis. Recent evidence indicates that natriuretic peptide precursor type C (NPPC) produced by mural granulosa cells stimulates the generation of cyclic guanosine 3′,5′-monophosphate (cGMP) by cumulus cell natriuretic peptide receptor 2 (NPR2), which diffuses into oocyte via gap junctions and inhibits oocyte phosphodiesterase 3A (PDE3A) activity and cyclic adenosine 3′,5′-monophosphate (cAMP) hydrolysis and maintains meiotic arrest with a high intraoocyte cAMP level. This cAMP is generated through the activity of the Gs G-protein by the G-protein-coupled receptor, GPR3 and GPR12, and adenylyl cyclases (ADCY) endogenous to the oocyte. Further studies suggest that endocrine hormones, such as follicle-stimulating hormone (FSH), LH, 17β-estradiol (E2) and oocyte-derived paracrine factors (ODPFs), participate in oocyte meiosis possibly by the regulation of NPPC and/or NPR2. A detailed investigation of NPPC and NPR2 expression in follicle cells will elucidate the precise molecular mechanisms of gonadotropins, and control the arrest as well as resumption of meiosis.     

Inhibition of LH-RH induced release of gonadotropins by substance P in cultured rat anterior pituitary cells]

Using anterior pituitary cells cultured for 7 days and then incubated for 4 hrs, substance P, an undecapeptide, inhibited the stimulatory effect of LH-RH on the release of LH-RH on the release of LH and FSH. This inhibitory effect, which was similar for both gonadotropins was only observed when the adenopituitary cells were put in culture at DI and Proestrus stages of the oestrous cycle. Furthermore substance P partly inhibited the basal release of FSH at DI and DII stages but did never affected that of LH.     

Comparison of the time courses of luteinizing hormone (LH) secretion rates during continuous stimulation by LH-releasing hormone (LH-RH) in vivo and in vitro

Summary The patterns of LH secretion during constant stimulation of the pituitary glands of estradiol-treated ovariectomized rats with a maximally stimulating amount of LH-RH in vivo and in vitro correspond with each other qualitatively and quantitatively. In vitro the changes with time of the LH secretion rate are somewhat retarded, especially the occurrence of desensitization.     

Comparison of the time courses of luteinizing hormone (LH) secretion rates during continuous stimulation by LH-releasing hormone (LH-RH) in vivo and in vitro

The patterns of LH secretion during constant stimulation of the pituitary glands of estradiol-treated ovariectomized rats with a maximally stimulating amount of LH-RH in vivo and in vitro correspond with each other qualitatively and quantitatively. In vitro the changes with time of the LH secretion rate are somewhat retarded, especially the occurrence of desensitization.     

The effect of gonadotrophins on the steroidogenesis, in the ovary and testis of gonadotrophin-deprived fresh water teleost, Cyprinus carpio.

Studies have been made on the effect of LH, FSH, and LH+FSH on the gonadal steroidogenesis in gonadotrophin-deprived common carp. LH alone and in combination with FSH was more effective than FSH in stimulation of steroidogenesis.     

Enhancement of serum FSH levels after pinealectomy

The effect of pinealectomy on the circadian periodicity in serum gonadotropins was investigated in adult male rats. Pinealectomy resulted in an elevation of the serum FSH concentration 7 days after the operation. A 4-fold increase in serum FSH over 24 h with no concommitant increase in serum LH following pinealectomy suggests that the control of FSH secretion could be mediated via the pineal gland.     

Enhancement of serum FSH levels after pinealectomy

Summary The effect of pinealectomy on the circadian periodicity in serum gonadotropins was investigated in adult male rats. Pinealectomy resulted in an elevation of the serum FSH concentration 7 days after the operation. A 4-fold increase in serum FSH over 24 h with no concommitant increase in serum LH following pinealectomy suggests that the control of FSH secretion could be mediated via the pineal gland.     

Neuroendocrine effects of a non-steroidal compound of testicular origin

Summary The effects of the compound (+)-1,4-diphenylbutane-2,3-diol (DPB, synthetized in the testes) on gonadotropin secretion have been studied in castrated male rats. DPB, when injected subcutaneously, does not modify serum levels of LH and FSH. On the contrary, the local implantation of DPB in the median eminence of the hypothalamus results in a significant elevation of serum FSH. It is suggested that DPB may play a physiological role in the control of FSH release.This work has been supported by grants of the Ford Foundation, New York, and of the CNR, Biology of Reproduction Program, Rome. Kits for LH and FSH radioimmunoassay have been kindly provided by the Rat Pituitary Hormone Distribution Program of the National Institutes of Arthritis, Metabolism and Digestive Diseases of the National Institutes of Health. Diphenylbutane was a kind gift of Dr R. Neher of the Ciba-Geigy Laboratories, Basel, to Dr Iturriza. Thanks are also due to Mrs Paola Assi Brunone for her skilful technical assistance.     

Plasma steroid and protein hormone concentrations in patients with benign prostatic hypertrophy and in normal men.

Serum levels of testosterone, FSH, LH and prolactin were measured by RIA in patients with BPH (benign prostatic hypertrophy) and in control men. The testosterone concentrations in the BPH group were significantly lower than that of the control group. Serum concentrations of FSH, LH, prolactin and estradiol, did not differ significantly in the 2 groups.     

Plasma steroid and protein hormone concentrations in patients with benign prostatic hypertrophy and in normal men

Summary Serum levels of testosterone, FSH, LH and prolactin were measured by RIA in patients with BPH (benign prostatic hypertrophy) and in control men. The testosterone concentrations in the BPH group were significantly lower than that of the control group. Serum concentrations of FSH, LH, prolactin and estradiol, did not differ significantly in the 2 groups.     

Similarity between the effects of suprachiasmatic nuclei lesions and of pinealectomy on gonadotropin release in ovariectomized,sulpiride-treated and melatonin-replaced rats

The aim of this study was to compare the effects of pineal indole treatments on LH and FSH release in pinealectomized and suprachiasmatic lesioned and ovariectomized rats rendered hyperprolactinemic by acute sulpiride treatment. pinealectomy or suprachiasmatic nuclei lesions in female rats both decreased plasma LH and FHS at 10, but not at 20 d after surgery, whereas the daily afternoon administration of melatonin effectively restored levels of both gonadotropins to control values. In ovariectomized rats, pinealectomy or suprachiasmatic nuclei lesions were ineffective in counteracting the high plasma levels of LH and FSH. However, sulpiride treatment in both pinealectomized and suprachiasmatic nuclei lesioned and castrated female rats significantly decreased the levels of LH and FSH, an effect which was counteracted by daily afternoon melatonin administration. Other pineal indoles tested, i.e., 5-hydroxy- and 5-methoxytryptophol, were ineffective in regulating gonadotropin levels. The results suggest that the pineal gland, through its hormone melatonin, can modulate gonadotropin secretion by acting on a dopamine mechanism independent of hypothalamic suprachiasmatic areas.     

The effect of gonadotrophins on the steroidogenesis,in the ovary and testis of gonadotrophin-deprived fresh water teleost,Cyprinus carpio

Summary Studies have been made on the effect of LH, FSH, and LH+FSH on the gonadal steroidogenesis in gonadotrophin-deprived common carp. LH alone and in combination with FSH was more effective than FSH in stimulation of steroidogenesis.Acknowledgment. K.K. is thankful to University Grants Commission, Govt. of India, for a fellowship. Tranks are due to Dr R.W. Bates, NIH Bethesda, for the supply of bovine LH and FSH, and to ICI England for the supply of methallibure.     

Increased responsiveness of the hypothalamic-pituitary axis to steroid feedback effects in ovariectomized rats treated neonatally with monosodium L-glutamate

Chronic ovariectomized rats treated neonatally with MSG showed reduced circulating concentrations of LH coupled with elevated hypothalamic LHRH stores. Despite the apparent loss of LHRH secretion, the small pituitary glands showed an increased density of LHRH receptors and normal responsiveness to the releasing hormone. The positive feedback effects of progesterone on LH release in oestrogen-primed animals was greatly exaggerated reflecting the build-up of hypothalamic LHRH stores without loss of pituitary responsiveness to LHRH.     

Differences in luteinizing hormone release stimulated in rat anterior pituitary cells by leukotriene C4 and by gonadotropin-releasing hormone in vitro

Continuous administration of leukotriene C4 (LTC4, 10(-10) M) to superfused rat anterior pituitary cells increased LH release for 40 min only, whereas in a parallel experiment gonadotropin-releasing hormone (GnRH, 10(-9) M) evoked a continuous increase in hormone secretion. In contrast to GnRH, LTC4 did not desensitize rat anterior pituitary cells. The secretory action resulting from the administration of LTC4 (10(-10) M) was abolished for 40 min after previous stimulation. The results documented the dual action of LTC4 on LH exocytosis.     

Adaptation of the pituitary gland to prolonged LRH stimulation

Summary With prolonged constant rate infusion of luteinizing hormone-releasing hormone (LRH), the LH secretion rate of the rat pituitary gland changes continuously until a steady state of relative desensitization has developed. Recovery from this state can occur independently from changes in the pituitary's LH content.