Biomedical subjects

H Vaudry

Publications and source records attributed to H Vaudry.

At least 523 records · Page 29Linked to original sources

PubMed1984

Lack of effect of dexamethasone on corticosteroid production in the amphibian.

Previous studies have demonstrated the presence of glucocorticoid receptors in the adrenal gland and suggested a direct inhibitory effect of glucocorticoids on the adrenal cortex. The present study was designed to investigate the possibility that dexamethasone could inhibit adrenal steroidogenesis in amphibia. For this study, we have applied a well-defined perifusion model, using interrenal fragments from frogs (Rana ridibunda). The amounts of corticosterone and aldosterone released in the effluent perifusate were radioimmunoassayed by means of specific antisera which did not cross-react with dexamethasone. Administration of dexamethasone (10(-6) and 10(-5M) did not significantly alter the basal secretion of corticosterone and aldosterone. In addition, dexamethasone (10(-5)M) did not modify the response of the interrenal gland to graded doses of ACTH. Finally dexamethasone did not inhibit the stimulation of corticosterone and aldosterone production induced by a single infusion of the angiotensin II analogue [Sar1-Val5] AII. From these results, it is concluded that in amphibia, chronic dexamethasone administration does not exert a direct suppressive effect on adrenal secretion.

Adrenal Cortex Hormones↗

PubMed1984

The effect of the antimineralocorticoid RU 28318 on aldosterone biosynthesis in vitro.

The effect of RU 28318, a specific and highly potent aldosterone antagonist on mineralocorticoid biosynthesis has been studied using a new in vitro model which combined three original features: (1) a very specific radioimmunoassay for aldosterone (2) a simplified perifusion system and (3) frog interrenal tissue which spontaneously produces high amounts of aldosterone. A dose-related inhibition of aldosterone production was observed for doses ranging from 10(-5) to 10(-3) M of RU 28318. The intermediate dose of 10(-4) M caused 71% inhibition of aldosterone production. Long term infusion of RU 28318 for 8 h led to a significant, stable and reversible inhibition of aldosterone production. In addition, we provide evidence that RU 28318 is capable of blocking the stimulation of aldosterone secretion induced by synthetic ACTH or by angiotensin II analogue. The present results demonstrate that RU 28318 is responsible for a significant and reversible inhibition of spontaneous, ACTH-induced and angiotensin II-induced aldosterone biosynthesis in vitro.

Adrenal Glands↗

PubMed1984

Decreased corticotropin-releasing factor-like immunoreactivity in rat intermediate and posterior pituitary after stalk section.

We determined the corticotropin-releasing factor(CRF)-like immunoreactivity by radioimmunoassay in intermediate and posterior pituitary lobes of stalk-sectioned and sham-operated control rats. The antigenic determinant read by the CRF antibody used was contained within the region of amino acids 26 (Gln) to 37 (Leu) of the molecule. Intermediate and posterior lobes of control rats contained similar amounts of CRF (591 +/- 78 and 487 +/- 34 pg/mg protein, respectively). The section of the pituitary stalk produced a marked decrease in CRF-like immunoreactivity in both structures. The CRF content on the intermediate lobe after stalk section was 62 +/- 17 pg/mg protein, a decrease of 90%, and that of the posterior lobe was 90 +/- 13 pg/mg protein, an 83% decrease. Our results suggest that most of the CRF-like immunoreactivity in the rat intermediate and posterior pituitary lobes is contained in nerve fibers of brain origin and support the hypothesis of a role of CRF in the release of intermediate and posterior lobe peptides.

Animals↗

PubMed1984

Comparative immunoelectron microscopic localization of corticotropin-releasing factor (CRF-41) and oxytocin in the rat median eminence.

Comparative ultrastructural localization of corticotropin-releasing factor (CRF) and oxytocin was performed in the rat median eminence of Long Evans and Brattleboro rats. The peroxidase-antiperoxidase technique used on serial ultrathin sections revealed CRF and oxytocin neurosecretory granule colocalization in the same fibers of the internal layer running towards the posterior pituitary. It is probable that both these peptides coexist in the same granules. In the Brattleboro rats, while genetically lacking vasopressin, CRF was nevertheless shown to be present. In these rats, as was demonstrated in the Long Evans rats, CRF distribution paralleled that of oxytocin only in the internal zone of the median eminence.

Animals↗

PubMed1984

Characterization and localization of vasoactive intestinal peptide receptors in the rat lung.

Experiments were performed to characterize vasoactive intestinal peptide (VIP) receptors in rat lung and to study their localization by means of light and electron microscope autoradiography. [125I]Iodo-VIP binding to lung homogenate was inhibited by synthetic VIP, [Val5]secretin, secretin, and 7-27 secretin with half-maximal inhibition (ID50) values of 1.62, 13, 121 nmol/liter and up to 0.1 mmol/liter, respectively. The relative potency of [Val5]secretin and 7-27 secretin to displace the tracer indicated that [125I]iodo-VIP binds to specific VIP receptors. Kinetic studies showed two sites of binding, a site of high affinity with equilibrium dissociation constant (KD) values of 0.32 +/- 0.09 nM and Bmax of 88.7 +/- 28.6 fmol/mg protein and a lower affinity binding site with a KD value of 23.0 +/- 2.9 mM and Bmax of 584 +/- 136.9 fmol/mg protein. The localization of VIP receptors was performed by radioautography both in vivo after iv injection of [125I]iodo-VIP and in vitro by the direct application of the radiolabeled peptide to cryostat sections of unfixed lung tissue. After iv injection of label, silver grains were detected only over alveoli. Electron microscopy indicated that radioactivity was mostly associated with alveolar capillaries. In vitro localization studies showed specific binding not only to alveoli but also to bronchi. These studies suggest that VIP can act at different sites and that the peptide could gain access to alveoli via the general circulation.

Animals↗

PubMed1984

Role of exogenous and endogenous prostaglandins in steroidogenesis by isolated frog interrenal gland: evidence for dissociation in adrenocorticotropin and angiotensin action.

The role of prostaglandins (PGs) in the regulation of glucocorticoid and mineralocorticoid production by the frog interrenal (adrenal) gland was studied in vitro by means of continuous perifusion. No change in corticosteroid production was observed upon perifusion with PGA1 or PGA2 (up to 10(-6) M). Conversely six other primary PGs, and the prostacyclin I2 (PGI2), significantly stimulated corticosteroidogenesis: the order of potency being PGE1 greater than PGI2 greater than PGE2 greater than PGB2 greater than PGB1 greater than PGF2 alpha greater than PGF1 alpha. The ability of the PGs to stimulate corticosterone and aldosterone production indicated that PGs may contribute to corticosteroid biosynthesis and that endogenous PGs could act as mediators for certain corticosteroid-stimulating factors. The essential role of endogenous PGs for spontaneous corticosteroid biosynthesis was demonstrated by the marked inhibition of corticosterone and aldosterone production induced by indomethacin (IDM) and meclofenamic acid, two specific inhibitors of cyclooxygenase which reduced significantly PG formation in the interrenal tissue. Stimulation of steroidogenesis by ACTH or cAMP was not affected by infusion of IDM or meclofenamic acid. In addition, ACTH did not modify the production of PGs by interrenal tissue. In contrast, the stimulatory effect of the angiotensin II analog [Sar1-Val5] AII, was markedly reduced in the presence of these two PG synthetase inhibitors and a direct stimulatory effect of the AII analog on PGE2 release was demonstrated. The effects of a supramaximal dose of [Sar1-Val5] AII could not be enhanced by PGE1 indicating that the corticosteroid response to angiotensin II is dependent upon endogenous PG synthesis, whereas the additivity of the stimulatory effects of PGE1 and a maximal dose of ACTH confirmed that PGs are not involved in the mechanism of action of ACTH. These results show that endogenous and exogenous PGs are implicated in corticosteroidogenesis in the frog. Since, in fish and amphibia the adrenocortical tissue is in close contact with the kidney (a major source of PGEs) these results may suggest a physiological relationship between the kidney and the interrenal gland in lower vertebrates. In addition, our results provide the first evidence that the action of angiotensin II on both corticosterone and aldosterone production may be mediated by PGs, whereas ACTH and cAMP stimulate corticosteroidogenesis through another mechanism. Thus, our data support the concept that ACTH and angiotensin II activate the early steps of corticosteroid biosynthesis via different pathways.

Adrenal Glands↗

PubMed1984

[The coexistence of neuropeptides and catecholamines in the adrenal gland. Research on paracrine effects on adrenal cortex cells].

The aim of the present review was to compare in mammals and amphibians the data concerning the presence of neuropeptides in the chromaffin cells and the possible action of these substances on adrenocortical cell function. Major homologies are to be found concerning the coexistence in chromaffin granules of catecholamines, Met-and Leu-enkephalins, and their precursor, proenkephalin A. However, the inhibitory action that might be exerted by enkephalins in vitro on corticosteroid production in mammalian adrenal gland, does not occur in amphibia. Dynorphin has been isolated in bovine adrenal medulla extracts; the presence of this opioid peptide has not been reported yet in amphibian interrenal tissue. All chromaffin cells of the frog interrenal gland contain VIP-like immunoreactivity whereas this neuropeptide is not contained in the adrenal medulla of mammals, exept in certain pheochromocytomas. In the frog, VIP, Metenkephalin and catecholamines are co-sequestered in the same chromaffin granules. In addition, synthetic porcine or chicken VIP stimulate in vitro the secretion of corticosteroids by frog interrenal fragments. In mammals, the steroidogenic action of VIP has been observed exclusively in tumor cell lines. The existence of somatostatin has been demonstrated in the human adrenal medulla and in pheochromocytomas, but not in amphibia. Somatostatin has been found to inhibit the response of adrenocortical cells to angiotensin II in mammals. A similar effect of somatostatin was not observed in amphibia. The coexistence of VIP and catecholamines in frog chromaffin granules and the stimulatory effect exerted by VIP on corticosteroidogenesis suggest that, in these animals, VIP may be co-liberated with noradrenaline during stress conditions, and thus may act locally on adrenocortical cells to stimulate corticosteroid secretion.

Adrenal Cortex↗

PubMed1983

Immunocytochemical localization of corticotropin-releasing factor-like immunoreactivity in the human hypothalamus.

In order to study the distribution of corticotropin-releasing factor-like immunoreactivity (CRF-LI) in the human hypothalamus, an immunocytochemical localization of this neurohormone was performed. Using antibodies developed against ovine CRF, we have localized CRF-LI in parvicellular neuronal cell bodies in the paraventricular nucleus. Immunostained fibers were observed in the median eminence, the pituitary stalk and the posterior pituitary. These results suggest that immunoreactive material related to ovine CRF is present in the human hypothalamus with a distribution similar to that observed in the rat.

Corticotropin-Releasing Hormone↗

PubMed1983

Coexistence of vasoactive intestinal peptide and enkephalins in the adrenal chromaffin granules of the frog.

The chromaffin cells of the frog adrenal gland were studied at the electron microscopic level by means of the immunoperoxidase technique. Using specific antibodies against Met-enkephalin, Leu-enkephalin and vasoactive intestinal peptide (VIP), the coexistence of the three neuropeptides in the chromaffin cells was demonstrated. Furthermore, all three peptides were co-located in the 200-400 nm dense-core vesicles which also stained for dopamine-beta-hydroxylase. These results, which suggest concomitant release of catecholamines and neuropeptides by the chromaffin cells, support the concept that VIP is involved in the stimulation of adrenal steroid secretion during stress conditions.

Adrenal Medulla↗

PubMed1983

Biological and immunological characterization of alpha-melanocyte-stimulating hormone (alpha-MSH) in two neuronal systems of the rat brain.

Recent immunocytochemical studies have demonstrated the existence of two different neuronal systems containing alpha-MSH-like material in the brain: one originating from the arcuate nucleus and the other one from the dorsolateral hypothalamus. The aim of the present study was to further characterize alpha-MSH in these two discrete regions of the rat diencephalon. Intracerebroventricular administration of colchicine resulted in a marked decrease in the number of ACTH and beta-endorphin nerve fibers and a significant reduction in ACTH and beta-endorphin content in the dorsolateral hypothalamus. Conversely, colchicine treatment did not alter alpha-MSH, ACTH or beta-endorphin content in the arcuate nucleus and did not significantly affect alpha-MSH concentration in the dorsal region. Reverse-phase high-performance liquid chromatography showed that the major alpha-MSH-like compound localized in the dorsal hypothalamus co-migrated exactly with synthetic alpha-MSH, whereas the arcuate nucleus contained 5 peptides cross-reacting with alpha-MSH antibodies, 4 of them being different from standard alpha-MSH. Significant amounts of biologically active melanotropin, which migrated on Sephadex G-25 columns like synthetic alpha-MSH, were also detected in both the arcuate nucleus and dorsolateral hypothalamus. Taken together, these data demonstrate that the alpha-MSH cell bodies located in the dorsolateral hypothalamus specifically produce authentic alpha-MSH, whereas the alpha-MSH cell bodies in the arcuate nucleus also contain ACTH, beta-endorphin and several peptides immunologically related but not identical to alpha-MSH.

Adrenocorticotropic Hormone↗

PubMed1983

Co-localization of vasoactive intestinal peptide (VIP) and enkephalins in chromaffin cells of the adrenal gland of amphibia. Stimulation of corticosteroid production by VIP.

Recent studies have shown that biologically active peptides and monoaminergic neurotransmitters coexist in certain neuronal cell populations. Using the immunofluorescence technique, we have examined the localization of enkephalins, vasoactive intestinal peptide (VIP) and tyrosine hydroxylase in the adrenal gland of the frog Rana ridibunda. Most chromaffin cells which stained for tyrosine hydroxylase contained VIP-like immunoreactivity, whereas methionine- (Met-) and leucine- (Leu-) enkephalin-like immunoreactivity was detected in about 40% of the cells revealed by the anti-tyrosine hydroxylase serum. No VIP- or enkephalin-like immunoreactive nerve fibres were observed. Since in the frog, the chromaffin cells are in close contact with the adrenocortical (interrenal) tissue, a possible action of VIP and opiates on corticosteroidogenesis has been investigated. At doses 10(-6) and 10(-5) M, 20-min infusions of synthetic porcine or chicken VIP elicited a significant increase in corticosterone and aldosterone production by perifused frog adrenals, in a dose-dependent manner. As compared to ACTH, VIP was several orders of magnitude less effective in stimulating corticosteroid production. Morphine, Met- and Leu-enkephalins (10(-5) M) had no effect on spontaneous secretion of corticosteroids. In addition, Met- and Leu-enkephalins (10(-5) M) did not alter the production of corticosterone induced by ACTH. THese results suggest that VIP contained in the chromaffin cells of the frog adrenal gland may exert a local action in stimulating corticosteroid production by the interrenal tissue.

Acetylcholine↗

PubMed1983

Immunocytochemical evidence of intragranular acetylation of alpha-MSH in the melanotrophic cells of the rabbit.

Melanotrophic cells of the pars intermedia of the rabbit were studied at the electron-microscopic level by means of the immuno-gold technique with the use of antisera against gamma 3-MSH and alpha-MSH. Both antibodies labelled all secretory vesicles stored in the peripheral cytoplasm, but secretory vesicles and intrasaccular condensations in the Golgi area reacting for gamma 3-MSH were not labelled with the antibody against alpha-MSH. Since this antibody appears to recognise only the acetylated form of alpha-MSH, the present observations suggest that acetylation occurs at a stage subsequent to the Golgi packaging, during maturation of the secretory vesicles. Thus, the morphological evidence supports biochemical data in favour of intragranular processing of opiomelanocortin in melanotrophic cells.

Acetylation↗

PubMed1983

In vitro study of frog (Rana ridibunda Pallas) neurointermediate lobe secretion by use of a simplified perifusion system. II. Lack of action of thyroxine on TRH-induced alpha-MSH secretion.

Thyrotropin-releasing hormone (TRH) stimulates alpha-melanocyte-stimulating hormone (alpha-MSH) secretion in amphibia as well as thyrotropin-stimulating hormone (TSH) and prolactin secretions in mammals. Since thyroid hormones regulate the stimulatory effect of TRH on TSH and prolactin, the possible role of thyroxine (T4) in the control of alpha-MSH secretion in amphibia, has been investigated. Neurointermediate lobes of Rana ridibunda were perifused in amphibian culture medium for 7 hr and the amounts of alpha-MSH released into the effluent perfusate were measured by radioimmunoassay. In vivo treatment with T4 (0.5 mg/kg twice a day for 9 days) did not modify the in vitro response of the neurointermediate lobes to TRH (10(-9) to 10(-7) M). In addition, prolonged infusion of T4 in vitro did not alter spontaneous and TRH-induced alpha-MSH release. In spite of the inhibitory effect of T4 on TRH-induced TSH and prolactin secretions in mammals, the present data show that, in frogs, thyroid hormone does not modulate the stimulation of alpha-MSH secretion induced by TSH.

Animals↗

PubMed1983

Immunological characterization of endorphins, adrenocorticotropin, and melanotropins in frog hypothalamus.

To gain more information about the nature and regulation of opiomelanocorticotropic peptides in the frog diencephalon, radioimmunological determinations of alpha- and beta-MSH. ACTH, beta- and gamma-endorphins have been performed in hypothalamic extracts. Sephadex G-50 gel filtration revealed a single peak of alpha-MSH-like immunoreactivity (alpha-MSH-LI) comigrating with synthetic alpha-MSH. Two peaks of ACTH-LI were observed; the major one eluting slightly before human ACTH. Using a porcine beta-endorphin antiserum which exhibited 45% cross-reaction (on a molar basis) with ovine beta-LPH, one major peak of beta-endorphin-LI and two additional components were observed in the elution profile; none of these peaks coeluted with synthetic porcine beta-endorphin. No significant beta-MSH or gamma-endorphin-LI was detected. To investigate a possible role of glucocorticoids on the level of opiomelanocorticotropic peptides in frog hypothalamus, 60 male frogs were treated with dexamethasone (300 micrograms/day) during 8 days. Dexamethasone treatment did not modify the chromatographic distribution and the total amount of alpha-MSH-LI and ACTH-LI in hypothalamic extracts. A slight (15%) increase in beta-endorphin-LI was even observed after 8 days of dexamethasone treatment. From these results it is concluded that three classes of opiomelanocorticotropic peptides are present in the frog hypothalamus in the following order of concentration: beta-endorphin-LI greater than alpha-MSH greater than ACTH. In addition, circulating glucocorticoids which significantly reduce the concentration of opiomelanocorticotropic hormones in the distal lobe of the frog pituitary (S. Jégou, M. C. Tonon, F. Leboulenger, C. Delarue, J. Côté, G. Pelletier, and H. Vaudry (1981a). In "Adv. Physiol. Sci.' E. Stark, G. B. Makara, Zs. Acs, and E. Endröczi, eds., Vol. 13, pp. 129-133. Pergamon, Budapest.) do not modify the amount of these peptides in the hypothalamus.

Adrenocorticotropic Hormone↗

PubMed1983

In vitro study of frog (Rana ridibunda Pallas) neurointermediate lobe secretion by use of a simplified perifusion system. III. Effect of neuropeptides on alpha-MSH secretion.

It has been previously demonstrated that thyrotropin-releasing hormone (TRH) stimulates in vitro the release of alpha-melanocyte-stimulating hormone (alpha-MSH) in frog. In the present study, the effects of various neuropeptides on spontaneous and/or TRH-induced alpha-MSH secretion were investigated, using a well-defined perifusion system technique. Vasoactive intestinal peptide, (VIP) a neurohormone which stimulates TRH target cells in mammals, was totally devoid of effect on frog melanotrophs although VIP-like material could be detected in neurointermediate lobe extracts. Somatostatin-like immunoreactive material was found in high concentrations in the frog neurointermediate lobe complex, but synthetic somatostatin (from 10(-10) to 10(-6) M) did not modify the spontaneous release of alpha-MSH. At doses of 10(-8) and 10(-6) M, synthetic somatostatin did not modify TRH-induced alpha-MSH secretion. Morphine (10(-5) M) and opioid peptides (10(-10) to 10(-6) M) had no effect on spontaneous alpha-MSH secretion. In addition, methionine enkephalin (10(-5) M) did not modify the stimulatory effect of TRH on alpha-MSH secretion. From these results we conclude that, among the neuropeptides which modulate prolactin secretion in mammals, only TRH is involved in alpha-MSH secretion in the frog.

Animals↗

PubMed1983

Effect of Tyr-Pro-Leu-Gly-NH2 on melanotropin-stimulating hormone release in the frog and rat.

The tetrapeptide Tyr-Pro-Leu-Gly-NH2 (Tyr-MIFI) has been recently characterized in the hypothalamus and pineal of the rat. Since the concentration of Tyr-MIFI in the brain is increased by pinealectomy and is higher when the animals are in the dark, the possibility that Tyr-MIFI could be a physiological regulator of melanotropin secretion has been investigated. For this study a well-defined perifusion model has been applied, using whole neurointermediate lobes from male frogs (Rana ridibunda Pallas) or male Wistar rats. The amount of alpha-MSH released in the effluent perifusate was measured by means of a sensitive and specific radioimmunoassay method. For concentrations ranging from 10(-10) to 10(-6) M, Tyr-MIFI did not significantly alter the spontaneous release of alpha-MSH in the frog nor did it alter the release of alpha-MSH in the rat. Since it has been recently demonstrated that the tripeptide pGlu-His-Pro-NH2 (mammalian TRH) is a specific MSH-releasing factor in the frog, the possibility that Tyr-MIFI could modulate the response of the intermediate lobe of the frog to TRH has also been investigated. No alteration of TRH-induced alpha-MSH release was observed in the presence of a 100-fold excess of Tyr-MIFI. In addition, Tyr-MIFI was found to be unable to lighten the skin of the frog (Rana pipiens) when applied directly to the pituitary of the darkened animals. Thus these results definitively rule out the possibility that Tyr-MIFI is the melanotropin-release inhibiting factor in the frog or rat.

Animals↗

PubMed1983

The frog pars intermedia contains only the non-acetylated form of alpha-MSH: acetylation to generate alpha-MSH occurs during the release process.

Pulse-chase experiments revealed that the frog pars intermedia synthesizes the desacetyl form of alpha-MSH. Its structure was shown to be similar, if not identical, to the mammalian structure. During release two additional peptides derived from desacetyl alpha-MSH appeared, one being alpha-MSH. We conclude that the N-acetylation of newly synthesized MSH is associated with release of the hormone. Radioimmunoassays and bioassays showed that the non-acetylated peptide is the only tissue form of MSH and confirmed that acetylation is linked to release.

Acetylation↗

PubMed1983

Comparative immunocytochemical localization of corticotropin releasing factor (CRF-41) and neurohypophysial peptides in the brain of Brattleboro and Long-Evans rats.

The localization of CRF-41 related peptide was studied in the brain and posterior pituitary of the homozygous rats for the inherited diabetes insipidus (Brattleboro strain, DI) and of the Long-Evans rats (LE) as control. It was compared to the distribution of vasopressin (AVP), oxytocin (OXY) and OXY-neurophysin (N I). In both strains, CRF-41 was identified in two morphologically distinct systems: one was a hypothalamoneurohypophysial system simultaneously containing CRF-41, OXY and N I; the other was a hypothalamoinfundibular system carrying CRF-41 only. CRF containing neurons were located in the periventricular area of the anterior hypothalamus, in the retrochiasmatic part of the supraoptic nuclei (SON) and, for some of them, in the antechiasmatic part of SON. CRF immunostainings were enhanced by colchicine treatment in LE rats and by DDAVP therapy in DI rats.

Animals↗