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H Vaudry

Publications and source records attributed to H Vaudry.

At least 451 records · Page 25Linked to original sources

Study of neuropeptide Y-containing nerve fibers in the human penis.

Neuropeptide Y 1-36 (IR-NPY) immunoreactive nerve-fiber processes have been observed in tunicae of veins and arteries and in smooth muscles of the human penis taken at autopsy or during surgery by use of light- and electron-microscopic immunohistochemical techniques. Numerous IR-NPY nerve fibers were mostly concentrated in the inner part of the adventitia close to the media of the arterial and venous vessels and among the intracavernous smooth muscle cells. IR-NPY nerve fibers were less abundant in veins than in arteries. Positive somata were not observed in the penises. At the ultrastructural level, IR-NPY were localized exclusively in large, dense granules of nerve terminals by means of the postembedding immunogold technique. In the deep dorsal vein, IR-NPY nerve fibers were also located in the media formed by an outer circular and an inner longitudinal layer. In the intracavernous and dorsal arteries, they showed the highest density in the inner part of the adventitia. In the corpora cavernosa and in the corpus spongiosum, IR-NPY nerve processes were intermingled between the smooth-muscle fibers around the sinusoid spaces. IR-NPY nerve fibers were present in the cavernous nerves close to the central arteries. The urethra did not show any IR-NPY-positive nerve fibers. This peculiar distribution of IR-NPY nerve fibers suggested that they could participate in regulating arterial and venous blood flow and intracavernous smooth-muscle tone. NPY may therefore be of importance in some of the mechanisms of penile erection especially during detumescence.

Adolescent↗

Effect of osmotic pressure on prolactin release in rainbow trout: in vitro studies.

To investigate a possible effect of osmotic pressure on prolactin (PRL) release in rainbow trout, we developed a technique for in vitro perifusion of trout pituitaries. Changes in osmotic pressure similar to those observed in fish plasma during transfer experiments did not induce significant modifications of PRL release. In contrast, high-amplitude variation of osmotic pressure resulted in clear modifications of PRL secretion: hyperosmotic medium caused a reduction in PRL release, while infusion of hyposmotic medium induced a transitory increase in PRL release. By using different concentrations of mannitol, we found that the modifications of prolactin secretion could not be ascribed to alterations of the ionic composition of the medium but actually resulted from variations in the osmotic pressure of the incubation medium. In further experiments osmotic pressure was decreased from 300 to 220 mOsm/kg or from 400 to 300 mOsm/kg; a similar transitory increase in PRL release was observed. Measurement of gonadotropin (GtH) in the perifusion effluent medium showed that PRL and GtH secretion followed similar patterns. Thus, our results suggest a possible mechanical effect of wide changes in osmotic pressure on pituitary cell membranes. These data indicate that the rainbow trout differs notably from nonsalmonid teleost species thus far studied in the lack of sensitivity of its PRL cells to osmotic pressure.

Animals↗

Biological, chromatographical, and radioimmunological evidence for a melanotropin-like peptide in the central nervous system of Locusta migratoria.

Our recent immunocytochemical study has demonstrated the existence of alpha-melanocyte stimulating hormone (alpha-MSH)-like material in the locust central nervous system. The aim of the present study was to further characterize alpha-MSH in the locust brain by its biological effect on frog skin and by high-pressure liquid chromatography in combination with radioimmunological and biological detection methods. Parallel radioimmunoassay (RIA) curves of crude nervous tissue extracts coupled with bioactivity in a very specific bioassay suggest similarity between the locust alpha-MSH-like substance and synthetic alpha-MSH. The highest concentration of alpha-MSH immunoreactive material in the central nervous system was found in the optic lobes, where alpha-MSH immunoreactive cell bodies are localized, as was previously shown by immunocytochemistry. High concentrations of alpha-MSH immunoreactive material were also detected in the thoracic ganglia of the locust ventral nervous system. The application of locust brain extracts to gel permeation HPLC resulted in a similar elution profile of the bioactive and immunologically active substances, both coeluting with synthetic alpha-MSH. As is the case for vertebrate brain material, reverse-phase HPLC revealed four alpha-MSH immunoreactive peaks. One of the peaks coelutes with monoacetyl-alpha-MSH and other RIA-positive material elutes at times close (but not identical) to the methionine sulfoxide forms of alpha-MSH. Peak three, however, elutes in a very different position from desacetyl-alpha-MSH. Peaks are absent in the position of desacetyl-alpha-MSH. Similarity between the locust alpha-MSH-related substance and authentic alpha-MSH is discussed.

Animals↗

Effect of atrial natriuretic factor on corticosteroid production by perifused frog interrenal slices.

In order to investigate a possible role of atrial natriuretic factor (ANF) in the control of corticosteroid biosynthesis in amphibians, we have examined the effect of synthetic ANF (Arg 101-Tyr 126) on perifused frog interrenal slices. ANF did not affect the spontaneous secretion of corticosterone and aldosterone. In contrast, ANF (10(-6) M) inhibited ACTH-and angiotensin II-stimulated corticosteroid production. ANF was more potent in suppressing aldosterone than corticosterone secretion. Immunocytochemical studies using a specific ANF antiserum revealed the presence of ANF-like immunoreactive fibers in the vicinity of interrenal cells. It is thus proposed that, in amphibians, both "hormonal" ANF secreted by myocytes and "neurohormonal" ANF delivered by peptidergic nerve terminals coursing among interrenal cells may partake in the regulation of corticosteroidogenesis.

Adrenal Glands↗

Interrenal activity in the female lizard Lacerta vivipara J.: in vitro response to ACTH 1-39 and to [Sar1, Val5] angiotensin II (ANG II).

A perifusion system technique was developed in order to determine in vitro the respective roles of ACTH and ANG II in the regulation of adrenal steroidogenesis in the lizard Lacerta vivipara. Synthetic human ACTH 1-39, administered as 20-min pulses, stimulated corticosterone (B) and aldosterone (A) release in a dose-dependent manner. The increase in corticosterone output was higher than that in aldosterone output, leading to an enhancement of the B/A ratio. Iterative stimulations with 1 nM ACTH (20-min pulses every 120 min) led to reproducible increases in corticosterone and aldosterone release. Prolonged stimulation with 1 nM ACTH (up to 240 min) caused a sustained increase in corticosteroid release, suggesting that, in the lizard, ACTH does not induce any desensitization phenomenon. The angiotensin II analogue [Sar1, Val5] ANG II also stimulated corticosterone and aldosterone release in a dose-dependent manner; the stimulatory effects of ANG II on both steroids were very similar. These results indicate that, in lizards, ACTH plays a major role in the regulation of adrenal steroidogenesis. Since ANG II stimulates the production of gluco- and mineralocorticoids, our data raise the question of the existence of two cell types synthesizing corticosterone and aldosterone, respectively, in reptiles.

Adrenal Glands↗

In vitro study of frog adrenal function--IX. Evidence against the involvement of lipoxygenase metabolites in the control of steroid production.

The possible role of arachidonic acid metabolites of the lipoxygenase pathway in the regulation of steroidogenesis was studied in vitro using perifused frog interrenal (adrenal) glands. Graded doses of arachidonic acid (10(-6)-10(-4)M) increased the production of corticosterone and aldosterone in a dose-dependent manner. In the presence of indomethacin (5 X 10(-6)M), the effect of arachidonic acid on steroid secretion was totally abolished. Nordihydroguaiaretic acid (NDGA: 10(-6)M), a lipoxygenase inhibitor, did not alter the spontaneous secretion of corticosteroids and did not impair the stimulatory effect of arachidonic acid. In the presence of NDGA, both ACTH and angiotensin II were still able to stimulate corticosteroid production. Our data support the view that arachidonic acid metabolites play an important role in the regulation of amphibian steroidogenesis. Moreover, the results show that the lipoxygenase pathway is not involved in the spontaneous secretion of corticosteroids and in angiotensin II- or ACTH-induced steroidogenesis.

Adrenal Glands↗

Effect of the intermediate filament inhibitor IDPN on steroid secretion by frog adrenal glands.

In order to determine the role of intermediate filaments in adrenal steroidogenesis, we have studied the effect of IDPN (beta-beta'iminodipropionitrile), an intermediate filaments perturbing agent, on corticosteroid secretion by frog interrenal glands in vitro. A 6-h administration of IDPN (10(-3) M) did not affect the spontaneous release of corticosterone and aldosterone. While IDPN did not alter the response of adrenal fragments to ACTH, the drug caused a marked decrease in angiotensin II-induced stimulation of corticosterone and aldosterone production. These results indicate that, in contrast to microfilaments, which play an important role in spontaneous steroidogenesis, intermediate filaments are not required for basal corticosteroid secretion but are involved in the mechanism of action of angiotensin in frog adrenocortical cells.

Actin Cytoskeleton↗

Serotonin stimulates corticosteroid secretion by frog adrenocortical tissue in vitro.

The mode of action of serotonin (5-HT) in the regulation of frog adrenal steroidogenesis was studied in vitro using the perifusion system technique. Graded doses of 5-HT (from 10(-8) to 10(-6) M) increased both corticosterone and aldosterone production in a dose-dependent manner. Short pulses (20 min) of 10(-6) M 5-HT, administered at 130 min intervals within the same experiment, did not cause any desensitization phenomenon. Indomethacin (IDM; 5 microM), a cyclooxygenase inhibitor which induced a dramatic decrease in the spontaneous secretion of corticosteroids, did not impair the stimulatory effect of 5-HT on corticosterone and aldosterone production. In the absence of calcium, 5-HT (10(-6) M) was still able to stimulate corticosteroid production. Dantrolene (5 x 10(-5) M), a blocker of calcium mobilization from intracellular pools which significantly inhibited the spontaneous production of corticosteroids, did not suppress 5-HT-evoked corticosteroid secretion. These results show that 5-HT, stored in adrenal chromaffin cells, may act as a paracrine factor to stimulate adrenal steroidogenesis in the frog. Our data also indicate that the mechanism of action of 5-HT does not depend on prostaglandin biosynthesis.

6-Ketoprostaglandin F1 alpha↗

Electron microscope immunocytochemical localization of thyrotropin-releasing hormone (TRH) prohormone in the rat hypothalamus.

Using an antiserum to a synthetic peptide corresponding to the sequence Phe-178-Ser-199 of the precursor of thyrotropin-releasing hormone (pro-TRH), we have studied by immunoelectron microscopy the ultrastructural localization of pro-TRH in the rat hypothalamus. In the paraventricular nucleus neuronal cell bodies, dendrites and axons were labelled. The dense core vesicles were strongly immunoreactive. Immunostained cell bodies, dendrites and endings were frequently observed in contact with unidentified neuronal elements. In the median eminence, numerous endings containing positive dense core vesicles were detected. These results suggest that pro-TRH and/or fragment(s) of pro-tRH could play a neuromodulator/neurotransmitter role in the hypothalamus and could also be released into the pituitary portal plexus.

Animals↗

Regional processing of the N- and C-terminal domains of proopiomelanocortin in monkey pituitary and brain.

The total content and extent of processing of the gamma 3MSH and beta-endorphin-containing N- and C-terminal domains of proopiomelanocortin were determined in the anterior and intermediate lobes of the pituitaries and in 11 regions of the brains of three Rhesus monkeys. Most immunoreactive gamma 3MSH and beta-endorphin was located in the pituitary lobes, although significant amounts were also found in several brain regions. Sephadex column chromatography revealed that gamma 3MSH immunoreactivity was found primarily as 4K and 9K forms; no gamma 1MSH was detected. beta-Endorphin immunoreactivity was found as beta-endorphin, beta-lipotropin, and as a 5K form which may represent beta-endorphin extended N-terminally by part or all of beta-MSH. In the anterior lobe of the pituitary, the predominant products were 9K gamma 3MSH and beta-lipotropin; in the intermediate lobe, more processed forms (4K gamma 3MSH, beta-endorphin and 5K beta-endorphin) appeared to be preferentially stored. The pattern of processing in various brain regions was similar to that of the intermediate lobe of the pituitary.

Animals↗

Distribution of atrial natriuretic factor-like immunoreactivity in the brain of the frog Rana ridibunda.

The localization of atrial natriuretic factor (ANF)-like immunoreactivity in the central nervous system of the frog Rana ridibunda was examined by the indirect immunofluorescence technique, using an antiserum against synthetic ANF (Arg101-Tyr126). Immunoreactive cell bodies were principally found in the dorsal and medial pallium, the medial septal nucleus, the ventrolateral and anteroventral areas of the thalamus, the lateral forebrain bundle, the posterolateral thalamic nuclei, the preoptic nucleus, the dorsal infundibular nucleus, and the anteroventral tegmentum nucleus of the mesencephalon. Numerous cell bodies and a very dense fiber bundle were visualized in the interpeduncular nucleus. All the areas mentioned above contained a high density of immunoreactive fibers. In addition, the amygdala, the infundibular nucleus, the median eminence, and most of the areas of the mesencephalon contained a moderate number of ANF-positive nerve processes. In the frog pituitary, fibers and nerve terminals were found in the peripheral zone of the neural lobe. The intermediate and anterior lobes of the frog pituitary were totally devoid of ANF immunoreactivity. These results indicate that ANF-like material is widely distributed in the frog brain and that ANF may be involved in various brain functions including neuroendocrine regulations.

Animals↗

Autoradiographic study of somatostatin receptors in the rat hypothalamus: validation of a GTP-induced desaturation procedure.

The radiolabelled somatostatin analogs [125I-Tyr0,DTrp8]S14 and [Leu8,DTrp22,125I-Tyr25]S28 were used as radioligands to study the distribution of somatostatin receptors in the rat hypothalamus. Previous studies have detected very few somatostatin-binding sites in the hypothalamus using in vitro autoradiography. Since the lack of autoradiographic labelling has been ascribed to the occupancy of the receptors by endogenous ligands, we have developed a method using guanosine triphosphate (GTP) pretreatment to unmask somatostatin receptors. Preincubation of brain slices with 10(-6) M GTP, by desaturating the occupied receptors, made it possible to reveal the wide distribution of somatostatin-binding sites in the rat hypothalamus. Somatostatin-14 binding site populations were observed in numerous hypothalamic areas including the preoptic area where the receptors likely account for self-inhibition of somatostatin release, the supraoptic nucleus, the bed nucleus of the stria terminalis, the anterior hypothalamic nucleus, the perifornical area, the zona incerta and a mediolateral area located laterally to the ventromedian and dorsomedian nuclei and limited laterally by the mammillo-thalamic tract, the fornix and the optic tract. All structures showing S14-binding sites were labelled by the S28 radioligand. In addition, the paraventricular parvocellular nucleus contained exclusively S28-binding sites, which could be involved in the inhibitory effect of S28 on CRF-mediated endocrine and sympathetic responses. A moderate density of S28-preferring sites was also detected in the periventricular nucleus. In summary, GTP preincubation of brain slices appeared to be a useful technique to reveal multiple somatostatin receptors populations in the brain. The widespread distribution of somatostatin receptors in the hypothalamus is in total agreement with the variety of physiological effects of the somatostatin peptide family.

Animals↗

Patch-clamp study of the ionic currents underlying action potentials in cultured frog pituitary melanotrophs.

The ionic conductance mechanisms underlying the action potential behaviour of frog melanotrophs in primary culture were studied by using the patch-clamp technique in whole-cell configuration. The action potentials spontaneously generated by these cells were predominantly sodium spikes with a calcium component. Voltage-dependent sodium, calcium, potassium and calcium-activated potassium currents were identified and analysed separately. The voltage-dependent sodium current was characterized by its fast kinetic, its low-threshold activation, its voltage-dependent inactivation and a tetrodotoxin sensitivity. Calcium currents were identified on the basis of their ionic selectivity to divalent cations (Ba2+, Ca2+, Co2+) and their time course. Only two of the three well-documented calcium currents could be detected in frog melanotrophs. A sustained calcium current (ICaS) and an inactivating calcium current (ICaN) were elicited by step depolarizations up to -20 mV. ICaN inactivated for membrane potentials more positive than -50 mV; its inactivation appeared to be both voltage- and calcium-dependent. Transient calcium current (ICaT) has never been observed. Two types of potassium currents were identified: voltage-dependent potassium (IKV) and calcium-activated potassium currents, (IK[Ca]). They were both suppressed by tetraethylammonium chloride, whereas only IK(Ca) was blocked by cobalt. These major ionic currents underlying spontaneous electrical activity are assumed to be involved in the process of alpha-melanocyte-stimulating hormone release. The present study provides the ground for future investigations regarding the relationships between the electrical and secretory activities in amphibian pars intermedia cells.

Action Potentials↗

Role of calcium in thyrotrophin-releasing hormone-stimulated release of melanocyte-stimulating hormone from frog neurointermediate lobe.

The effect of modifications of extracellular calcium concentrations on alpha-MSH release has been studied using perifused frog neurointermediate lobes. Increasing concentrations of calcium (from 2 to 10 mmol/l) gave rise to a dose-related stimulation of alpha-MSH secretion, whereas reduction of Ca2+ from 2 to 1.5 mmol/l partially inhibited alpha-MSH release. The direct effect of extracellular Ca2+ on alpha-MSH secretion was confirmed by the dose-dependent stimulation of alpha-MSH release induced by the calcium ionophore A23187. Perifusion with a calcium-free medium or blockade of Ca2+ channels by 4 mmol Co2+/l both resulted in an inhibition of spontaneous and TRH-induced alpha-MSH release. Conversely, administration of verapamil or methoxyverapamil (10 mumol/l each) did not alter basal secretion and had no effect on the response of the glands to TRH. Nifedipine (10 mumol/l), which was able to block KCl (20 mmol/l)-evoked alpha-MSH release, induced a slight inhibition of basal alpha-MSH secretion, indicating that extracellular Ca2+ levels may regulate alpha-MSH release in part by Ca2+ influx through voltage-dependent Ca2+ channels. In contrast TRH-induced alpha-MSH release was not affected by nifedipine or dantrolene (10 mumol/l), and BAY-K-8644 (1 mumol/l) did not significantly modify the response of neurointermediate lobes to TRH. Taken together, these results suggest that TRH-induced alpha-MSH secretion is associated with calcium influx across the plasma membrane and that calcium entry caused by TRH may occur through nifedipine/verapamil-insensitive Ca2+ channels.

Animals↗

Involvement of the cytoskeleton in the steroidogenic response of frog adrenal glands to angiotensin II, acetylcholine and serotonin.

In order to determine the role of the cytoskeleton in adrenal steroidogenesis, we have studied the effect of cytochalasin B (a microfilament-disrupting agent) and vinblastine (an antimicrotubular drug) on corticosteroid secretion by frog interrenal tissue in vitro. Perifusion of interrenal fragments with cytochalasin B (50 mumol/l) induced a marked inhibition of basal corticosteroid output. In addition, stimulation of corticosteroidogenesis by all corticotrophic factors was also inhibited by cytochalasin B. Using an immunohistochemical technique and specific anti-tubulin antiserum, we verified that vinblastine (10 mumol/l) was responsible for the disappearance of the microtubular network in adrenocortical cells. Administration of vinblastine (10 mumol/l) did not affect the spontaneous secretion of corticosterone and aldosterone and had no effect on the steroidogenic response of interrenal glands to angiotensin II and acetylcholine. In contrast, vinblastine was responsible for a marked decrease in serotonin-induced stimulation of corticosteroid production. On the other hand, data from high-performance liquid chromatography showed that infusion of cytochalasin B or vinblastine was not associated with the production of any new steroid which could interfere in the radioimmunoassays. Taken together, these data suggest that microfilaments are involved in a late and common step of corticosteroidogenesis while microtubules are only required for the coupling of the secretory response to certain corticotrophic factors such as ACTH and serotonin.

Acetylcholine↗

Formation of 11 beta-hydroxysteroids requires the integrity of the microfilament network in adrenocortical cells.

In order to determine the role of microfilaments in adrenal steroidogenesis, we have studied the effect of cytochalasin B, a microfilament-disrupting agent, on the kinetics of [3H] pregnenolone conversion to labelled metabolites by frog interrenal tissue in vitro. Cytochalasin B (5 x 10(-5)M) induced a 50 to 70% decrease in corticosterone, 18-hydroxycorticosterone and aldosterone biosynthesis while the formation of progesterone and 11-desoxycorticosterone was not affected. These results suggest that microfilaments interfere in the conversion of 11-desoxycorticosterone to corticosterone probably by controlling the movement of 11-desoxycorticosterone from the reticulum to the mitochondria.

Actin Cytoskeleton↗

Hypothalamic alpha-melanocyte-stimulating hormone (alpha-MSH) is not under dopaminergic control.

A possible dopaminergic regulation of hypothalamic proopiomelanocortin (POMC)-containing neurons has been investigated in rats by means of in vivo and in vitro approaches. Acute or 3-weeks chronic in vivo treatments with the dopaminergic agonists apomorphine (1 mg/kg: s.c.) and 2-Br-alpha-ergocriptine (2.5 mg/kg; s.c.) or the dopaminergic antagonist haloperidol (0.15-3 mg/kg; i.p.) had no significant effect on the concentration of alpha-melanocyte-stimulating hormone (alpha-MSH) in two hypothalamic regions: arcuate nucleus (AN) and dorsolateral area (DLH). In the same way, chronic administration of the dopaminergic agonists or antagonist did not induce any change in hypothalamic contents of beta-endorphin, another peptide derived from POMC. Reverse-phase high-performance liquid chromatographic analysis revealed that acetic acid extracts of AN and DLH both contained two major forms of alpha-MSH-like peptides: deacetylated alpha-MSH and authentic alpha-MSH. The ratio between these two forms was not altered after acute haloperidol treatment (3 mg/kg, i.p.). The possible effect of dopamine on the release of hypothalamic alpha-MSH was studied in vitro using perifused rat hypothalamic slices. Infusion of dopamine (10(-7)-10(-5)M) or its antagonist haloperidol (10(-5)M) had no effect on spontaneous alpha-MSH release from hypothalamic tissue. In addition, none of these drugs had any effect on potassium (50 mM)-induced alpha-MSH release. It is concluded that dopaminergic neurons are not involved in the regulation of synthesis, post-translational processing (acetylation) or release of hypothalamic alpha-MSH.

Animals↗

Melanin-concentrating hormone (MCH) is colocalized with alpha-melanocyte-stimulating hormone (alpha-MSH) in the rat but not in the human hypothalamus.

Melanin-concentrating hormone (MCH)-containing neurons have recently been localized in the dorsolateral region of the rat hypothalamus, an area where the second alpha-MSH system is found which contains only alpha-MSH and none of the pro-opiomelanocortin (POMC)-related peptides. In order to study the morphological relationships between the MCH and alpha-MSH neuronal systems, we have studied the immunocytochemical localization of both MCH and alpha-MSH in the rat hypothalamus. The same study was also performed in the human hypothalamus where there is only one alpha-MSH system which contains alpha-MSH as well as the other POMC-related peptides (first alpha-MSH system). In the rat dorsolateral hypothalamus, we could demonstrate that most neuronal cell bodies stained for MCH also contained immunoreactive alpha-MSH. In the human hypothalamus, neuronal cell bodies stained for MCH were observed only in the periventricular area whereas cell bodies containing alpha-MSH were exclusively located in the infundibular (arcuate) nucleus. In the rat, immunoelectron microscopy showed labelling for MCH in the dense core vesicles of positive neurons and double-staining techniques clearly demonstrated that both immunoreactive MCH and alpha-MSH could be consistently detected in the same dense core vesicles. These ultrastructural studies then suggest that these two peptides should be released simultaneously from neurons located in the rat dorsolateral hypothalamus.

Animals↗