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C Bugnon

Publications and source records attributed to C Bugnon.

At least 37 records · Page 2Linked to original sources

[Specific hybridization of a synthetic oligonucleotide in rat hypothalamic neurons immunoreactive to immune serum against salmon melanin-concentrating hormone].

An oligonucleotide probe corresponding to the 9 C-terminal residues encoded by a complementary DNA of a rat peptide related to salmon melanin concentrating hormone (MCH) was synthetized. It specifically hybridized to the neurons stained by antisera to MCH in the rat posterior hypothalamus, as seen by coupling in situ hybridization and immunocytochemical methods. This result validates our sequence determination. This oligonucleotide will be useful to establish the complete sequence of the rat MCH precursor molecule. It will also constitute a valuable tool to study physiological or experimentally-induced changes in the expression of the rat MCH gene.

Animals↗

[Cloning of the cDNA, while encoding a hypothalamic neuropeptide in the rat, related to the melanin-concentrating hormone in the salmon].

In order to identify the neuropeptide related to salmon melanin-concentrating hormone (MCH) synthetized by neurons of the posterior hypothalamus in the mammals, we have screened rat hypothalamus and rat brain cDNA expression libraries using MCH antiserum. Five recombinants were isolated, which cDNAs were amplified using the polymerase chain reaction. One of them hybridized to RNAs exclusively located in hypothalamic neurons stained by the same antiserum, as seen by performing in situ hybridization and immunocytochemical techniques on the same section. The sequence analysis showed that this cDNA corresponds to the end of the open reading frame encoding a MCH-like peptide and to the 3' untranslated region. The rat MCH is very similar to salmon MCH (greater than 85% homologies in the 14C-terminal residues).

Amino Acid Sequence↗

[Cloning and sequence analysis of cDNA of common precursors of three hypothalamic neuropeptides immunologically related to human somatocrinin 1-37, alpha melanotropin and salmon melanin-concentrating hormone].

We have cloned and sequenced DNAs complementary to the mRNA encoding the precursor of the rat melanin-concentrating hormone. This allowed us to elucidate the primary structure of the 96 C-terminal residues of this precursor. It contains three possible sites for enzymatic cleavage enabling the generation of MCH and of two additional neuropeptides. These three peptides can bind one of our antisera raised against human somatocrinin 1-37, alpha-melanotropin and salmon melanin-concentrating hormone, which immunocytochemically stain the same neuron population in the hypothalamus.

Amino Acid Sequence↗

Growth hormone-deficient dwarfism in the rat: a new mutation.

Mutations in animals have provided insight into many aspects of normal and pathological human physiology. This paper reports the discovery and initial characterization of a new mutant dwarf rat. The mutation, inherited as an autosomal recessive, arose spontaneously in a breeding colony of Lewis rats at the Medical Research Council Cellular Immunology Unit, Sir William Dunn School of Pathology, Oxford, U.K., in 1985 and the strain has now been established both in Oxford and at Mill Hill. Body growth in the mutant is retarded such that at 3 months of age both males and females weigh approximately 40% less than their normal litter-mates, and continue to grow at a slower rate. The mutants show a selective reduction in pituitary GH synthesis and storage (pituitary GH concentrations were approximately 10% of normal in males and 6% in females). The concentration of their anterior pituitary trophic hormones (LH, TSH, prolactin and ACTH) were within the normal range in dwarf animals. Exogenous GH treatment for 5 days resulted in an increase in growth rate from 1.5 +/- 0.3 to 3.9 +/- 0.4 g/day in male mutants, and 0.8 +/- 0.2 to 3.1 +/- 0.1 g/day in females. Longitudinal bone growth rates were more than doubled by this treatment from 49 +/- 5 to 100 +/- 10 micron/day in females and from 52 +/- 11 to 131 +/- 16 micron/day in males. Dot blot and Northern blot analysis of pituitary mRNA extracts revealed that the GH message in mutants was between 20 and 25% of normal, and that the GH transcript was of normal size.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Unrelated peptide immunoreactivities coexist in neurons of the rat lateral dorsal hypothalamus: human growth hormone-releasing factor1-37-, salmon melanin-concentrating hormone- and alpha-melanotropin-like substances.

Antisera raised against 3 unrelated synthetic neuropeptides - salmon melanin-concentrating hormone, human growth hormone-releasing factor1-37, and alpha-melanotropin - stained the same extensive neuron population in lateral and dorsal areas of the posterior hypothalamus. Controls for specificity have shown that these 3 antisera bind 3 different epitopes. Differences in intracellular staining patterns suggest that these epitopes could be borne by distinct peptides.

Animals↗

Human corticoliberin hypothalamic neuroglandular system: comparative immunocytochemical study with anti-rat and anti-ovine corticotropin-releasing factor sera in the early stages of development.

Development of the paraventriculo-infundibular corticoliberin system was studied by immunocytochemical analysis of human hypothalamic sections using antisera raised against rat or ovine corticotropin-releasing factor (CRF). This comparative study confirms the presence of a significant number of CRF-immunoreactive fibers in the median eminence during the 16th week of fetal development and suggests they may appear as early as the 14th week. Some hypothalamic peri- and paraventricular neurons, observed from the 12th week, are rat-CRF-immunoreactive but not ovine CRF-immunoreactive. There appears to be chronological differences concerning the ability of the two antisera to recognize hypothalamic structures during the early stage of development.

Animals↗

[Contribution of immunocytochemistry to the study of the development of neuroglandular peptidergic systems in the human fetal hypothalamus].

Immunohistochemistry makes possible the in situ detection of neuropeptides in the cell bodies were they are synthesized, in the fibers that carry them, and in endings. Immunohistochemistry appears necessary to identify and map peptidergic neurons and to study their ontogeny. From 1975, we have carried the immunohistochemical study of several hypothalamic neuronal populations in the human fetus: LH-RH (1976), somatostatin (1977), pro-opiocortin (1978), vasopressin and oxytocin (1979), corticoliberin (1982), somatocrinin (1983), and hypothalamic neurons containing an unidentified peptide (1984). Comparative ontogenetical studies have also been performed in rats.

Animals↗

[Ontogenetic data on the peptidergic interneuronal population in immunoreactivity of the GRF 37 type serum of the human posterolateral hypothalamus. Immunocytochemical studies using anti-GRF 37 and anti-MCH (melanin-concentrating hormone) immune sera].

Human posterolateral hypothalamic neurons are revealed with an anti GRF 37 serum as soon as the 7th week of fetal life. The same neuronal population can be observed in the adult brain even in hypothalami from old subjects, with the same distribution, and similar immunoreactivity than in fetal stages. These neurons are revealed using a melanin concentrating hormone (MCH) antiserum; the MCH immunoreactivity appears at the same stage of fetal development than GRF 37 immunoreactivity. The two antisera recognize two epitopes on one or two molecules. Those new facts agree with an hypothesis about the very important and permanent functional role of that new human hypothalamic interneuronal system.

Adult↗

Coexpression of human growth hormone-releasing factor 1-37-like and alpha-melanotropin-like immunoreactivities in neurones of the rat lateral dorsal hypothalamus.

Antibodies recognizing the 29-37 sequence of the human somatocrinin specifically stain a large population of interneurones located in the lateral dorsal hypothalamus. Staining comparisons revealed that these perikarya also contain alpha-MSH-like immunoreactivity. The neurones exhibiting human GRF1-37-like immunoreactivity correspond to the system previously shown to present alpha-MSH-like and rat CRF-like immunoreactivities.

Animals↗

[Ontogenesis in man, of a population of neurons in the dorsal and lateral hypothalamus, secretors of a peptide that has not yet been characterized].

A human GRF 1-37 antiserum demonstrates a new neuronal system in lateral perifornical areas of the human hypothalamus. The molecule that is revealed in those neurons cannot be somatocrinin. Perikarya are abundant. They are observed beginning with the 9th week of development. alpha-MSH-like immunoreactivity, which is showed in the analogous cells of the rat, is not yet established in human. The early differentiation of those neurons and their abundance during the fetal life attest to the important neurophysiological function of the unidentified peptide they secrete.

Fetus↗

Immunohistochemical demonstration of a new neurone system in rat brain using antibodies against human growth hormone-releasing factor (1-37).

An antiserum raised against the 1-37 N-terminal sequence of the human growth hormone-releasing factor (GRF) stains two distinct neurone systems in rat brain: the somatocrinin neuroglandular system in the infundibulum and a large, hitherto unknown population of interneurones, the perikarya of which are chiefly distributed in the lateral hypothalamus. Their projections are abundant in several hypothalamic and limbic areas. These interneurones contain high amounts of a substance, probably a new neuropeptide, sharing an epitope with the 29-37 sequence of the human GRF.

Animals↗

Anatomical and ontogenetic studies of the human paraventriculo-infundibular corticoliberin system.

In human fetus, newborn, infant and adult hypothalami, antibodies to ovine corticoliberin-41 stain a paraventriculo-infundibular neuroglandular pathway. The perikarya are located in the paraventricular nucleus, they mainly project to the ventral and lateral areas of the median eminence. Eminential corticoliberin-positive fibres appear during the 16th week of fetal life, and increase in number during the following weeks. Perikarya were first revealed in the 19th week. In some areas of the median eminence, corticoliberin-, vasopressin- or [Met]enkephalin-immunoreactive terminals are similarly distributed. Sequential stainings or staining comparison of contiguous semi-thin sections failed to prove the coexpression of corticoliberin and [Met]enkephalin immunoreactivities in fibres, but indicated that corticoliberin and vasopressin immunoreactivities may be coexpressed in a few fibres. Those methods enabled us to observe, in the paraventricular nucleus, perikarya revealed by corticoliberin and vasopressin antisera. Our results suggest a possible release of corticoliberin in portal vessels of the median eminence beginning in the 16th week of fetal life, i.e. 8 weeks later than appearance of the corticotrophs in the pituitary. Establishment of a corticoliberin hypothalamic control of pituitary corticotrophs at mid gestation agrees with previous physiological and teratological studies. Abundance, as well as immunostaining intensity of the corticoliberin processes, in the infant and adult median eminence attest to the physiological importance of this system. Close vicinity of corticoliberin, vasopressin and [Met]enkephalin fibres, in some eminential areas and coexpression of corticoliberin and vasopressin immunoreactivities in some neurons, are morphological correlates of functional relations which were reported.

Adult↗

[Coexpression of immunoreactivity with an anti-corticoliberin immune serum and immunoreactivity with an anti-vasopressin immune serum in human hypothalamic neurons].

Confrontations of immuno-stainings as well as sequential stainings with corticoliberin and vasopressin antisera were performed on contiguous sections of human median eminence (ME) or hypothalamic paraventricular nucleus (PVN). Results show that a double immunoreactivity to both IS is expressed in some parvicellular PVN perikarya, and suggest that such a double immunoreactivity can be possible in some ME fibers. Similar results had been previously obtained in animals only under experimental conditions.

Corticotropin-Releasing Hormone↗

Corticoliberin neurons: cytophysiology, phylogeny and ontogeny.

In the rat hypothalamus, antibodies to ovine CRF41 stain neurons of a paraventriculo-infundibular neuroglandular pathway. CRF like immunoreactivity (CLI)-containing perikarya are mostly packed in the parvocellular division of the paraventricular nucleus. Their morphology and topography differ from that of other peptidergic neurons. However a few CLI perikarya are also stained with vasopressin antibodies. CLI neurons project massively to the external layer of the median eminence (ELME). Adrenalectomy induced a total depletion of ELME CLI 12 to 24 h after surgery, followed by a secondary accumulation already conspicuous 5 days later. This biphasic evolution, identical to that of ELME vasopressin, is totally prevented by a replacement therapy with dexamethasone. Reserpine also induces an acute depletion of ELME CLI and vasopressin, that can be prevented by a monoamine oxidase inhibitor pretreatment. These results indicate the involvement of CLI neurons in the corticotropic axis, suggesting that they are indeed corticoliberin neurons. Among the extrahypothalamic locations of CLI neurons their abundance in the amygdala central nucleus is of interest since it is involved in the corticotropic axis. A similar pattern of CLI was noticed in several mammalian brains and also in lower vertebrates (birds, reptiles, amphibians, fishes). Species adaptations of CLI neurons were observed: CLI neurons are of the cerebrospinal fluid contacting type in the turtle. CLI fibres terminate close to corticotrophs in the fish pituitary. This suggests a direct excitosecretory role of CRF on these cells and concurs with a CRF function of CLI peptide even in fishes. CLI processes and terminals appear in the human fetal ELME at the 16th week of development and increase in number during the following weeks. Perikarya are seen at 19 weeks. In the rat CLI fibers and perikarya were detected as early as the 18th day of fetal development. Thus, paraventriculo-infundibular CLI system develops later than corticotrophs. This chronology perfectly concurs with the results of previous physiological and experimental studies.

Adrenalectomy↗

The CRF neuron: immunocytochemical study.

In the central nervous systems of several species belonging to different vertebrate classes, immunocytochemical stainings with an antiserum to ovine CRF 41 show multiple location of CRF perikarya and various areas containing CRF fibres and terminals. These stainings reflect species and interspecies functional adaptations of the CRF neurones which constitute a prominent hypothalamo-infundibular system involved in pituitary gland control, and also interneurone systems as attested to by extrahypothalamic perikarya and by CRF perisomatal endings in several brain areas. Perikarya of the hypothalamo-infundibular system are mainly packed in the paraventricular nucleus (mammals, birds) or in homologous areas, e.g.: paraventricular organ (turtle) where they are CSF-contacting neurones and preoptic nucleus (amphibians, fishes). In all species but fish, CRF fibres end in the median eminence (ME) against portal vessels. In fish, CRF processes terminate in the peripheral areas of proadenohypophyseal neurodigitations, close to corticotrophs. In all species these stainings are abolished by preabsorption of the serum by CRF. In fishes, reptiles and amphibians they are also suppressed by urotensin I, which is thought to be the teleost's CRF. Adrenalectomy experiments in the rat provided evidence for a corticosteroid regulation of ME CRF: short term (12-24 hr) adrenalectomy induces a complete depletion of CRF immunoreactivity followed by a secondary accumulation (5-20 days). This biphasic evolution is prevented by a dexamethasone replacement therapy. Inhibitory role of catecholamines on CRF release was indicated by: disappearance of ME CRF, induced by a single reserpine injection and suppression of this effect by monoamineoxidase inhibitor (pargyline or tranylcypromine) pretreatment. CRF fibres were first observed in the ME at the 16th week of fetal development in the human, and at the 18th day in the rat fetus. Thus, immunoreactive CRF system develops later than pituitary corticotrophs.

Adrenalectomy↗

Ontogeny of the neuroglandular system revealed with HPGRF 44 antibodies in human hypothalamus.

Using the indirect immunofluorescence technique, an antiserum to synthetic hpGRF 44 (the 44 residue peptide with high growth hormone releasing activity isolated from a human pancreatic tumor), antibodies of which interact with 40-44 C terminal amino acid sequence, stains a novel neuron system in the adult human hypothalamus. HpGRF-like immunoreactivity (GLI) containing neurons are different from previously known peptidergic systems. GLI perikarya are few in number and located mainly in the infundibular nucleus. In the median eminence, abundant GLI fibers terminate against portal capillaries in the same areas as somatostatin fibers which are morphologically different. No GLI staining was observed in hypothalami before 18 weeks of fetal development. GLI perikarya and intraeminential nerve terminals were simultaneously detected in 18-week-old fetuses attesting to a possible release of this peptide in the portal vessels as early as this stage. GLI neuron system was steadily revealed in 19- to 30-week-old fetuses. It was not detectable in 6 out of the 7 studied hypothalami from newborn infants. In most infant and adult hypothalami, it is well developed. Results presented here support the hypothesis (1) that hpGRF antiserum stains the neuron system producing the somatocrinin and (2) that this neuron system may begin in controlling pituitary somatotrophs at the 18th week of fetal development, which is in agreement with previous hypotheses.

Adult↗

Reserpine-induced depletion of corticoliberin (CRF)-like immunoreactivity in the zona externa of the rat median eminence.

An acute reserpine treatment has the same selective and marked depleting effect on corticoliberin-like immunoreactivity as on vasopressin-like immunoreactivity in the rat zona externa of the median eminence. Somatostatin and gonadoliberin immunoreactivities appear unmodified. Reserpine effect is blocked by pretreatment with monoamine oxidase inhibitors (pargyline or tranylcypromine). Present results support the notion of an inhibitory role of monoamines, particularly catecholamines, on the release of corticoliberin.

Animals↗

Immunocytochemical demonstration of a novel peptidergic neurone system in the cat brain with an anti-growth hormone-releasing factor serum.

In 15-day-, 1-month- and 1.5-month- old cats an antiserum against hpGRF-44 stained a novel hypothalamo-infundibular neuroglandular system. Its intra-eminential processes present an important development between 15 and 30 days of age. Perikarya and nerve fibres, which are abundant in several brain areas as early as the 15th day, suggest that the peptide revealed by hpGRF-44 antibodies also acts as a neuromodulator.

Animals↗