Biomedical subjects

B Breton

Publications and source records attributed to B Breton.

At least 37 records · Page 2Linked to original sources

PubMed1997

Effects of steroids on GTH I and GTH II secretion and pituitary concentration in the immature rainbow trout Oncorhynchus mykiss.

Using specific radio-immunoassays for rainbow trout GTH I and GTH II, the effects of testosterone and estradiol 17 beta have been studied or reinvestigated on the regulation of the secretion and the synthesis of the these two pituitary gonadotropins in the immature rainbow trout. After steroid implantation, the GTH II pituitary concentration is stimulated by testosterone and estradiol 17 beta for the entire period during which the plasma levels of these hormones are maintained to values comparable to those measured in the adult vitellogenic female rainbow trout. On the other hand, only testosterone induced a transient increase in the GTH I pituitary content 15 days after implantation, and estradiol provoked a decrease at day 30. The secretion of both GTH I and GTH II is stimulated by testosterone but not by estradiol 17 beta. Altogether, these results show that in the immature rainbow trout, testosterone preferentially modifies GTH I secretion, but not that of GTH II. They confirm that the stimulation of GTH II accumulation after testosterone or estradiol treatment would correspond to a stimulation of hormone synthesis. They evidence a differential action of both steroids on the synthesis of the two gonadotropins, especially a possible inhibition of GTH I synthesis by estradiol. They let suppose that the regulation of GTH I synthesis would involve factors other than steroids.

Animals↗

PubMed1997

Ivermectin and moxidectin in two filarial systems: resistance of Monanema martini; inhibition of Litomosoides sigmodontis insemination.

Effects of ivermectin and moxidectin were compared on two filarial species: Monanema martini which presents dermal microfilariae and induces Onchocerca-like lesions in its natural murid host Lemniscomys striatus, and Litomosoides sigmodontis (= L. carinii). M. martini microfilariae showed an unusual resistance to ivermectin, in vitro and in vivo; moxidectin was no more efficient. However, the two drugs used at high concentrations deeply altered the uterine embryogenesis, but had no lethal effect on adult filariae. L. sigmodontis blood microfilariae showed a great susceptibility to moxidectin, similar to that previously described for ivermectin. The two drugs also induced a long term effect because they inhibited the insemination of the female filariae. This result reinforces the observations made by other authors on the human parasite, Onchocerca volvulus.

Animals↗

PubMed1996

Steroid activation of the brain-pituitary complex gonadotropic function in the triploid rainbow trout Oncorhynchus mykiss.

Testosterone and estradiol were implanted into triploid rainbow trout, and their effects on brain and pituitary salmon gonadotropin-releasing hormone (sGnRH) content and on gonadotropin (GtH2) levels in the pituitary and blood were studied. In the hypothalamic areas, neither steroid altered sGnRH content. In the brain tissues (ventral telencephalon and preoptic areas), testosterone (0.5 mg/kg BW) significantly increased sGnRH contents and larger doses induced a more rapid increase. Estradiol induced a similar increase which occurred later than the response to the same dose of testosterone. Both steroids also stimulated pituitary sGnRH, with estradiol being less potent. Only testosterone induced long-term increases in plasma GtH2 content. Both steroids may act on the synthesis and release of GnRH, with testosterone having a greater influence on the control of gonadotropin secretion, especially at the end of the cycle.

Animals↗

PubMed1995

Do gonadotrophin-releasing hormone neurons express estrogen receptors in the rainbow trout? A double immunohistochemical study.

A double immunocytochemical procedure, with two different chromogens, was used to compare the respective distributions of estrogen receptor-immunoreactive cells and gonadotrophin-releasing hormone-immunoreactive neurons on the same sections of the brains of adult male and female rainbow trout (Oncorhynchus mykiss). Estrogen receptor-immunoreactive cells were observed in the ventral and lateral telencephalon, the preoptic region, the mediobasal hypothalamus, and the ventromedial thalamic nucleus. Gonadotrophin-releasing hormone-immunoreactive perikarya were detected in the olfactory bulbs, the ventral telencephalon, the preoptic area, and the mediobasal hypothalamus. Double-staining studies showed that, although some estrogen receptor-positive cells were in close proximity to gonadotrophin-releasing hormone-immunoreactive perikarya, careful examination of 550 gonadotrophin-releasing hormone-positive cells from five adult females and two adult males failed to demonstrate any evidence that gonadotrophin-releasing hormone neurons coexpress estrogen receptor in the brain of the rainbow trout. The present study provides, for the first time in teleosts, morphological evidence that gonadotrophin-releasing hormone neurons do not represent major direct targets for estradiol, suggesting that the positive feedback effects of estradiol onto the gonadotrophin-releasing hormone system are likely to be conveyed via other cell populations.

Animals↗

PubMed1994

Localization of salmon gonadotropin-releasing hormone mRNA and peptide in the brain of Atlantic salmon and rainbow trout.

The decapeptide gonadotropin-releasing hormone (GnRH) is a key hormone for the central regulation of reproduction. The distribution of salmon GnRH (sGnRH), which is the major form in salmonids, has been studied in different fish species by immunocytochemistry. Discrepancies in data concerning the distribution of sGnRH perikarya led us to investigate this problem in two species, the Atlantic salmon and the rainbow trout, with in situ hybridizaiton of sGnRH messenger, a highly specific molecular tool. By Northern blot analysis, the rainbow trout sGnRH messenger appears to be about 500 bases in length, which is close to those isolated from Atlantic salmon or masu salmon and characterized previously. In situ hybridization with riboprobes generated with Atlantic salmon sGnRH cDNA demonstrated that sGnRH perikarya are restricted to the ventral part of olfactory bulbs, telencephalon, and preoptic area. They are distributed on a nearly continuous line extending from the olfactory bulbs to the preoptic area in both salmonid species studied. Despite the presence of GnRH-like immunoreactivity in the preoptic magnocellular nucleus (NPOm) and in the tegmentum of the midbrain (MT), the sGnRH mRNA is not present in these two structures. Stained cells in NPOm could be target cells for GnRH and immunoreactive neurons in MT are likely to be chicken GnRH-II containing cells. Our study not only gives a precise distribution of the sGnRH system in two salmonids, Atlantic salmon and rainbow trout, but also clarifies the ambiguous data published up to now in rainbow trout.

Animals↗

PubMed1993

Solubilization and purification of the gonadotropin (GTH II) receptor from rainbow trout (Oncorhynchus mykiss) ovaries.

The rainbow trout (Oncorhynchus mykiss) ovarian gonadotropin (GTH II) receptor was solubilized by extraction with the nonionic detergent 1% Triton X-100 in the presence of 20% glycerol. The hormone-binding characteristics of the soluble receptors were similar to those of membrane-bound receptors: the Scatchard plot of the equilibrium binding data produced a straight line, suggesting that the solubilized GTH II receptors, like membrane-bound receptors, contained a single class of high affinity 125I-sGTH II binding sites with an association constant of 2-5 x 10(10) M-1 (Ka = 1.4-2 x 10(10) M-1 for membrane-bound receptor). The maximal binding capacity was very low and varied from 7 to 17 fmol/mg proteins (about 5 fmol/mg ovarian membrane protein). The soluble receptor was purified by a simple and rapid immunoaffinity chromatography. The sGTH II-solubilized receptor complex was adsorbed to anti-sGTH II beta-subunit gammaglobulins covalently linked to Sepharose 4B and then eluted with an acidic buffer. About 50% of the binding activity present in the Triton X-100 extract was recovered in the pH 4 eluate. The other binding sites were eluted as a hormone-receptor complex and/or a damaged form. The free purified receptor presented a Ka of 1.3 x 10(10) M-1 in agreement with those found in membrane preparation and solubilized extract.

Animals↗

PubMed1993

Pituitary as a target organ for toxic effects of P4501A1 inducing chemicals.

We report here that cytochrome P4501A1 in the male rainbow trout pituitary is highly inducible by beta-naphthoflavone. Pituitary cells containing inducible P4501A1 were identified by double immunostaining as gonadotrophs containing gonadotropin II. Thus, the pituitary gonadotrophs may be target cells for polyaromatic hydrocarbons. Elevated plasma levels of gonadotropin II (GTH II) and testosterone in the induced fish indicated that the functioning of the pituitary was disturbed. Because GTH II regulate the final stage of sexual maturation the results implies that exposure to P4501A1 inducing compounds may disturb this development stage.

Animals↗

PubMed1992

In vitro bioactivities of various forms of GnRH in relation to their susceptibility to degradation at the pituitary level in the rainbow trout, Oncorhynchus mykiss.

In vitro potencies of native and modified forms of salmon and mammalian gonadotropin-releasing hormone (GnRH) were studied in relation with their susceptibility to degradation by intact pituitary cells maintained in culture. The kinetics of degradation and the origin of the proteases involved in this process were examined. All the molecules tested (native and modified forms) were equipotent at doses between 10(-6) and 10(-7) M in inducing GtH release by cultured pituitary cells. On the other hand, their effectiveness differed at 10(-9) and 10(-8) M leading to the establishment of the following hierarchy of bioactivity: the native forms, LHRH and sGnRH, were the less potent, the fish analogues (DAla6Pro9Net)sGnRH and (DArg6Pro9Net)sGnRH were the more potent, and mammalian analogues with substitutions at position 6 and/or 10 were intermediate in potency. The native form sGnRH was weakly degraded while no degradation of the modified molecules was observed. The degradation of the native sGnRH occurred after 12 and 24 hr of incubation and the results indicate that the peptidases involved are released from the cells into the incubation medium.

Animals↗

PubMed1991

Neuropeptide Y stimulates in vivo gonadotropin secretion in teleost fish.

The effects of the neuropeptide Y (NPY), alone or in combination with a gonadotropin-releasing hormone analogue, D-Ala6-desGly10-Pro9-Net LHRH (LHRHa), have been studied on the in vivo secretion of the maturational gonadotropin (GtH2) in the rainbow trout Oncorhynchus mykiss and the common carp Cyprinus carpio. Depending on the species, two routes of administration were used: in trout, intraperitoneal injection (20 micrograms/kg body wt); in carp, direct infusion (3 micrograms/kg body wt) into the third ventricle via a temporary brain cannula. In both cases NPY alone induced a twofold increase in GtH2 secretion and peaked 2 to 4 hr administration regardless of the route of injection. The plasma gonadotropin levels returned to basal within 8 hr. The relative increases (peaked secretion/basal secretion) did not differ with the route of injection. When the animals were first treated with NPY and then LHRHa (20 micrograms/kg) 1 hr later, the magnitude of the response to LHRHa was greater in the animals pretreated with NPY, indicating either a potentiation of LHRHa action by NPY or additive effects of the two peptides. The return to basal levels also took longer in fish receiving NPY first. NPY may act directly at the pituitary level or activate central neuromediatory systems.

Animals↗

PubMed1991

Neuropeptide-Y in the trout brain and pituitary: localization, characterization, and action on gonadotropin release.

Using a specific antiserum raised against synthetic neuropeptide-Y (NPY), the distribution of NPY-like immunoreactivity in the brain and pituitary of the trout Oncorhynchus mykiss has been examined with the indirect immunofluorescence and peroxidase-antiperoxidase methods. The highest density of NPY-immunoreactive elements was found in the basal telencephalon and hypothalamus. In particular, NPY-immunoreactive neurons were located in the nucleus entopeduncularis and the preoptic nucleus. NPY-immunoreactive fibers were observed throughout the trout brain. The preoptic nucleus, the suprachiasmatic nucleus, and the nucleus entopeduncularis were densely innervated. In addition, NPY-positive fibers were detected in the nucleus lateralis tuberis and in the distal and intermediate lobes of the pituitary. The NPY-like peptide of the trout brain was characterized by combining HPLC analysis and radioimmunological detection. Serial dilutions of trout hypothalamus and pituitary extracts produced displacement curves that were parallel to the standard curve. HPLC analysis resolved a major peak which was slightly less hydrophobic than porcine NPY. The possible effect of NPY, either alone or in combination with a GnRH antagonist, on gonadotropin (GtH) release from trout pituitaries was investigated using a perifusion system technique. Graded concentrations of synthetic NPY induced a dose-dependent stimulation of GtH release. The stimulatory activities of NPY and various short chain analogs on GtH release were compared: the order of potency was NPY greater than NPY-(2-36) greater than NPY-(16-36) greater than NPY-(25-36). This result suggests that the biological determinant of NPY is located in the C-terminal part of the molecule. Administration of a short pulse of NPY or GnRH (10(-7) M each) induced a marked stimulation of GtH release. Prolonged infusion of the GnRH antagonist D-Phe2-6,Pro3-GnRH induced a significant reduction of GnRH-evoked GtH secretion. In addition, the GnRH antagonist blocked NPY-induced GtH release. The widespread distribution of NPY in the trout brain suggests the involvement of this neuropeptide in a variety of physiological functions. The present data support the view that NPY, released by nerve terminals in the distal lobe of the pituitary, may act presynaptically on GnRH fibers to modulate GtH release.

Animals↗

PubMed1990

A rapid and sensitive ELISA for rainbow trout maturational gonadotropin (tGtH II): validation on biological samples; in vivo and in vitro responses to GnRH.

A rapid and sensitive heterologous enzyme-linked immunosorbent assay (ELISA) was developed to measure rainbow trout maturational gonadotropin. Purified salmon maturational gonadotropin (sGtH II) was used as reference hormone. Optimization of the procedure was performed by using an anti-beta sGtH serum. Two procedures were developed: an equilibrium assay (which did not involve a preincubation step) which lasted for 8 hr and a nonequilibrium assay (which involved a preincubation step) which lasted for 26 hr. The nonequilibrium assay gave the best sensitivity (70 pg/ml sample). GtH II measurements on in vivo and in vitro samples from GnRH analogs or sGnRH experiments showed that the ELISA procedure could be used over a wide range of concentrations. The method was validated by comparing GtH II concentrations measured by both RIA and ELISA.

Animals↗

PubMed1990

A presumptive role for GABA in the stimulatory effects of Des-Gly10, [D-Ala6]-LHRH-ethylamide and pimozide on the gonadotropin release in carp.

To investigate the effect of endogenous gamma-aminobutyric acid (GABA) on the blood maturating gonadotropin (GtH) levels, or to study its interaction with pimozide (dopamine antagonist) and a luteinizing hormone-releasing hormone analog (LHRH-a), sexually mature male and female carps were treated with drugs that may either inhibit GABA biosynthesis or GABA degradation. In females the irreversible inhibitor of GABA-transaminase, gamma-vinyl GABA (GVG), which was to increase the endogenous GABA-ergic tone, had no influence on GtH release. On the other hand, the increased GtH response to the combination of pimozide (PIM) and LHRH-a was clearly enhanced by the administration of 3-mercaptopropionic acid (MPA), an inhibitor of the rate limiting enzyme of GABA-biosynthesis. In males the GABA-ergic compound, valproic acid (DPA) decreased LHRH-a stimulated GtH levels. In male carps that received PIM to diminish the dopaminergic inhibition of GtH release, the spermiating response to LHRH-a was increased by administration of MPA. These data suggest that GABA interacts with the action of dopamine and the gonadotropin releasing hormone (GnRH) on the release of GtH.

3-Mercaptopropionic Acid↗

PubMed1990

Calcium ions as a mediator in GnRH action on gonadotropin release in the common carp (Cyprinus carpio L).

Collagenase-dispersed carp pituitary cells in a perifusion system were used to study the role of calcium ions in the mechanism of GnRH action on the release of maturational gonadotropin (GtH) in fish. The specific calcium chelator EGTA and the calcium antagonist manganese (Mn2+) caused a 40% inhibition in the basal GtH release and completely blocked GnRH-stimulated GtH release. Short-term application of graded doses of calcium ionophore A23187 caused a dose-dependent increase in GtH secretion. A23187 failed to stimulate GtH secretion in the presence of EGTA. Depolarization of the membrane by K+ caused a strong stimulation of GtH release similar to the action of GnRH. Stimulatory action of K+ was inhibited by EGTA. These data suggest a role for extracellular calcium as an intracellular mediator in GnRH-stimulated, as well as in basal, GtH release in carp. The stimulation of GtH release by K+ also indicates that voltage-dependent processes could be involved in this phenomenon.

Animals↗

PubMed1990

Involvement of voltage-dependent calcium channels (VDCC) in the action of GnRH on GtH release in common carp (Cyprinus carpio L): comparison with K+ action.

The involvement of different types of voltage-dependent calcium channels (VDCC) in the stimulatory action of GnRH (in comparison with K+) on maturational gonadotropin (GtH) release was investigated using superfused carp pituitary cells. The action of these 2 stimulants was not modified either by D600 or nifedipine (drugs blocking L-type of VDCC). Cadmium (Cd2+), which blocks all types of VDCC indifferently, provoked a dose-dependent stimulation of GtH secretion. Cd2+ action was not altered by addition of sGnRH in any of the doses. Similar results were obtained using K+ as a secretagogue, but only the highest dose of Cd2+ (200 mumol/l) was able to completely block K+ action. Low doses (0.1 and 1 mumol/l) of the L-type VDCC activator BAY-K8644 did not change basal GtH secretion and had no effect on sGnRH-stimulated GtH secretion. Surprisingly, doses (10 mumol/l and higher) of BAY-K8644 evoked dose-dependent inhibition of GtH secretion. On the other hand, a higher concentration (20 mumol/l) of nifedipine provoked a stimulation of GtH release. Our results indicate that the stimulatory action of GnRH and K+ involves activation of a certain type of cadmium-sensitive VDCC (probably T- or N-type VDCC) whereas dihydropyridine and diphenylalkylamine sensitive VDCC (L-type VDCC) does not participate in this phenomenon. The inhibitory action of BAY-K8644 and, on the other hand, the stimulatory action of nifedipine indicate that L-type VDCC probably play a role in other physiological pathways regulating GtH release in carp.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗

PubMed1990

Neuropeptide Y: localization in the central nervous system and neuroendocrine functions.

Neuropeptide Y (NPY) is a 36-amino acid peptide first isolated and characterized from porcine brain extracts. A number of immunocytochemical investigations have been conducted to determine the localization of NPY-containing neurons in various animal species including both vertebrates and invertebrates. These studies have established the widespread distribution of NPY in the brain and in sympathetic neurons. In the rat brain, a high density of immunoreactive cell bodies and fibers is observed in the cortex, caudate putamen and hippocampus. In the diencephalon, NPY-containing perikarya are mainly located in the arcuate nucleus of the hypothalamus; numerous fibers innervate the paraventricular and suprachiasmatic nuclei of the hypothalamus, as well as the paraventricular nucleus of the thalamus and the periaqueductal gray. At the electron microscope level, using the pre- and post-embedding immunoperoxidase techniques, NPY-like immunoreactivity has been observed in neuronal cell body dendrites and axonal processes. In nerve terminals of the hypothalamus, the product of the immunoreaction is associated with large dense core vesicles. In lower vertebrates, including amphibians and fish, neurons originating from the diencephalic (or telencephalic) region innervate the intermediate lobe of the pituitary where a dense network of immunoreactive fibers has been detected. At the ultrastructural level, positive endings have been observed in direct contact with pituitary melanotrophs of frog and dogfish. These anatomical data suggest that NPY can act both as a neurotransmitter (or neuromodulator) and as a hypophysiotropic neurohormone. In the rat a few NPY-containing fibers are found in the internal zone of the median eminence and high concentrations of NPY-like immunoreactivity are detected in the hypothalamo-hypophyseal portal blood, suggesting that NPY may affect anterior pituitary hormone secretion. Intrajugular injection of NPY causes a marked inhibition of LH release but does not significantly affect other pituitary hormones. Passive immunoneutralization of endogenous NPY by specific NPY antibodies induces stimulation of LH release in female rats, suggesting that NPY could affect LH secretion at the pituitary level. However, NPY has no effect on LH release from cultured pituitary cells or hemipituitaries. In addition, autoradiographic studies show that sites for 125I-labeled Bolton-Hunter NPY or 125I-labeled PYY (2 specific ligands of NPY receptors) are not present in the adenohypophysis, while moderate concentrations of these binding sites are found in the neural lobe of the pituitary. It thus appears that the inhibitory effect of NPY on LH secretion must be mediated at the hypothalamic level.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1989

Development of an enzyme-linked immunosorbent assay for goldfish gonadotropin.

An enzyme-linked immunosorbent assay (ELISA) for goldfish gonadotropin (GTH) was developed with the intent of devising a simple, reliable and nonradioisotopic assay for the measurement of GTH in goldfish biological samples. In this assay, soluble GTH of the standards or samples competes with carp GTH (cGTH) immobilized on a solid support (96-well microplate) for the fixation on antibodies to the beta-subunit of carp gonadotropin. The immobilized antigen-antibody complexes are then revealed by the peroxidase-antiperoxidase (PAP) technique. After revelation of the peroxidase activity, the absorbance value of each well is measured with a microplate reader. The cGTH concentration used for coating the wells is 2 ng/ml and the final dilution of the specific antibody is 1:80,000. The assay can be performed within 24 h and can be used over a range of 0.125-4 ng/ml. At about 50% binding, the intra- and interassay coefficients of variation are 5% and 9% respectively. The displacement curves generated by goldfish plasma or pituitary perifusion fractions were strictly parallel to the standard cGTH. In addition, the stimulation by salmon gonadotropin-releasing hormone of pituitary fractions perifused in vitro caused an immediate increase in the GTH measured in the collected fractions, strongly reinforcing the assumption that this assay indeed measures GTH.

Animals↗

PubMed1987

Increased gonadotropin levels in goldfish do not result in alterations in circulating thyroid hormone levels.

To determine whether increases in gonadotropin levels are capable of altering thyroid function in goldfish, plasma thyroid hormone levels were measured following induced changes in endogenous gonadotropin secretion and injection of carp gonadotropin. Radio-frequency lesions placed in the nucleus preopticus periventricularis or monosodium-L-glutamate-induced lesions of the posterior nucleus lateralis tuberis (NLT) of the hypothalamus were capable of stimulating significant increases in plasma gonadotropin levels, but were without effect on plasma triiodothyronine (T3) or thyroxine (T4) at time intervals ranging from 5 hr to 10 days. Likewise, injections of a superactive analog of gonadotropin-releasing hormone resulted in profound increases in gonadotropin levels without associated changes in thyroid hormones. No changes in the circulating levels of T4 or T3 were observed in response to injection of purified carp gonadotropins whereas injection of bovine thyrotropin or carp pituitary extracts stimulated significant increases in T4. Radiofrequency lesions of the pituitary stalk or of the anterior NLT also resulted in significant increases in circulating levels of T4, but not of T3, at 10 and 30 hr postlesion. These results demonstrate that direct acute stimulation of circulating thyroid hormone levels is not an intrinsic action of endogenous goldfish gonadotropin and that activation of the reproductive system, leading to ovulation in some cases, is without effect on blood total thyroid hormone levels. Additionally, these results confirm that hypothalamic inhibition of the pituitary-thyroid axis exists in this teleost fish and demonstrate that interruption of this inhibition results in a time-dependent, high-magnitude increase in circulating thyroxine levels.

Animals↗

PubMed1986

A reinvestigation of the Gn-RH (gonadotrophin-releasing hormone) systems in the goldfish brain using antibodies to salmon Gn-RH.

The organization of Gn-RH systems in the brain of teleosts has been investigated previously by immunohistochemistry using antibodies against the mammalian decapeptide which differs from the teleostean factor. Here, we report the distribution of immunoreactive Gn-RH in the brain of goldfish using antibodies against synthetic teleost peptide. Immunoreactive structures are found along a column extending from the rostral olfactory bulbs to the pituitary stalk. Cell bodies are observed within the olfactory nerves and bulbs, along the ventromedial telencephalon, the ventrolateral preoptic area and the latero-basal hypothalamus. Large perikarya are detected in the dorsal midbrain tegmentum, immediately caudal to the posterior commissure. A prominent pathway was traced from the cells located in the olfactory nerves through the medial olfactory tract and along all the perikarya described above to the pituitary stalk. In the pituitary, projections are restricted to the proximal pars distalis. A second immunoreactive pathway ascends more dorsally in the telencephalon and arches to the periventricular regions of the diencephalon. Part of this pathway forms a periventricular network in the dorsal and posterior hypothalamus, whereas other projections continue caudally to the medulla oblongata and the spinal cord. Lesions of the ventral preoptic area demonstrate that most of the fibers detected in the pituitary originate from the preoptic region.

Animals↗