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Biomedical subjects

J Guy

Publications and source records attributed to J Guy.

At least 109 records · Page 6Linked to original sources
PubMed1987

Specific binding sites for prolactin and growth hormone in the adult rabbit lung.

Specific prolactin (PRL) and growth hormone (GH) binding sites were identified and characterized in lung membranes from male and female adult rabbits. The binding of iodinated human GH ([125I]iodo-hGH) and iodinated ovine PRL ([125I]iodo-oPRL) was time, temperature and protein dependent and was found to conform to the requirements defining a physiological receptor, in terms of hormonal and immunological specificities as well as kinetic properties. [125I]Iodo-hGH was displaced from lung membranes by hGH, oPRL, ovine GH and rat GH, while [125I]iodo-oPRL was effectively displaced only by oPRL and hGH. Scatchard plots of the competition curves of [125I]iodo-hGH and [125I]iodo-oPRL were both linear, suggesting, in each case, a single class of binding sites with affinity constants (Ka) of 1.74 +/- 0.64 X 10(9) M-1 and 0.78 +/- 0.28 X 10(9) M-1 and binding capacities of 6.43 +/- 0.53 and 4.16 +/- 0.69 fmol/mg protein, respectively. Anti-PRL-receptor antiserum significantly inhibited the binding of the [125I]iodo-oPRL to rabbit lung membranes, while it was less potent in preventing the binding of [125I]iodo-hGH, which has both lactogenic and somatogenic activity. Removal of endogenous ligand by treating lung membranes with 4 M MgCl2 increased specific binding of hGH about 2.5-fold, exposing additional specific binding sites without significantly changing the binding affinity. The level of binding of hGH and oPRL to rabbit lung did not show a pronounced sex differentiation. In summary, PRL and GH binding sites have been demonstrated for the first time in adult rabbit lung membranes, and they support the possibility of a physiological role for PRL and GH in the lung.

Animals↗
PubMed1987

[Pro-opiomelanocortin neuronal systems].

Proopiomelanocortin (POMC) is a glycoprotein which serves as a multihormonal precursor for corticotropin (ACTH), lipotropins (beta and gamma-LPH), melanotropins (alpha, beta- and gamma-MSH) and endorphins (alpha-, beta- and gamma-endorphins). This precursor protein is primarily synthesized in corticotrophs of the anterior lobe and in melanotrophs of the intermediate lobe of the pituitary, as well as in other organs or tissues such as the genitourinary tract, the gastrointestinal tract and leukocytes. POMC is also present in the central nervous system (CNS) and numerous studies have been conducted to determine the localization, biosynthesis and functions of POMC-derived peptides. The identification of POMC-neuronal systems has been achieved by combining immuno histochemical studies, biochemical analysis, bioassays and radioimmunoassays. Three groups of perikarya containing various POMC-related peptides have been identified. One of these is located in the arcuate nucleus in the basal hypothalamus and projects towards the septum, thalamus and telencephalon. Some fibers originating from the arcuate nucleus terminate in the nucleus of the solitary tract in the brainstem where a second group of POMC-containing nerve cells are located. The latter innervates both the mesencephalon, the brainstem and the spinal cord. A third group of neurons, which contain alpha-MSH but not other POMC-derivates, has been identified in the zona incerta in the dorso-lateral hypothalamus. Processing of POMC in the cell bodies of the arcuate nucleus follows a similar pattern as in the pituitary intermediate lobe. Endopeptidases called "acid-thiol-arginyl-proteases" cleave the prohormone at paired basic amino acids. The basic residues remaining on the resulting peptides are subsequently eliminated by the joint action of the less specific B-type carboxypeptidases and B-type aminopeptidases. alpha-MSH and beta-endorphin are among the major end products. Enzymatic modifications including N-alpha-acetylation by opiomelanotropin-acetyltransferase (OMAT) and/or C-terminal amidation by peptidyl-glycine alpha-amidating monooxygenase (PAM) occur after proteolytic processing. However, the rate of acetylation observed in hypothalamic POMC neurons is much lower than in the melanotrophs of the pars intermedia. Acetylation of MSH and endorphin is crucial in determining the biological potency of these peptides. Desacetyl alpha-MSH is far less active than alpha-MSH (monoacetyl alpha-MSH), whereas acetylated beta-endorphin has no opiate activity. The mechanisms regulating the activity of POMC-containing neurons are still unknown.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1986

Neuropeptide Y in the intermediate lobe of the frog pituitary acts as an alpha-MSH-release inhibiting factor.

The presence of neuropeptide tyrosine (NPY) in the intermediate lobe of the frog pituitary was demonstrated using indirect immunofluorescence, the immunogold technique and a specific radioimmunoassay combined with high pressure liquid chromatography (HPLC). A high density of NPY-containing fibers, was found among the parenchymal cells of the intermediate lobe. These fibers originated from the ventral infundibular nucleus, travelled via the median eminence to the pars intermedia. At the electron microscopic level, NPY-like material was found exclusively in nerve fibers where the product of the immunoreaction was associated to dense-core vesicles. High concentrations of NPY-like peptide were found in neurointermediate lobe extracts. After Sephadex G-50 gel filtration the major peak of immunoreactive material appeared to co-elute with synthetic porcine NPY. Conversely, HPLC analysis revealed that the NPY-like peptide of the frog pituitary had a retention time shorter than the porcine NPY. The localization of NPY-like material in the pars intermedia suggested a possible role of NPY in the regulation of melanotropic cell secretion. In fact, graded concentrations of synthetic NPY induced a dose-dependent inhibition of alpha-melanotropin (alpha-MSH) release in vitro. The lack of effect of a dopaminergic antagonist on NPY-induced alpha-MSH release inhibition demonstrated that the local dopaminergic system could not account for the NPY action. These results indicate that NPY located in the hypothalamo-hypophyseal system of the frog may act as a melanotropin-release inhibiting factor.

Animals↗
PubMed1986

Localization and identification of alpha-melanocyte-stimulating hormone (alpha-MSH) in the frog brain.

The distribution of alpha-melanocyte-stimulating hormone (alpha-MSH) in the central nervous system of the frog Rana ridibunda was determined by immunofluorescence using a highly specific antiserum. alpha-MSH-like containing perikarya were localized in the infundibular region, mainly in the ventral hypothalamic nucleus. A rich plexus of immunoreactive fibers directed towards the ventral telencephalic region was detected. Reverse-phase high-performance liquid chromatography and radioimmunoassay were used to characterize alpha-MSH-like peptides in the frog brain. Chromatographic separation revealed that immunoreactive alpha-MSH coeluted with synthetic des-N alpha-acetyl alpha-MSH, authentic alpha-MSH and their sulfoxide derivatives. The heterogeneity of alpha-MSH-like material in the frog brain was in marked contrast with the figure observed in the intermediate lobe of the pituitary gland where only des-N alpha-acetyl alpha-MSH is present. These findings support the existence of discrete alpha-MSH immunoreactive neurons in the frog brain containing both desacetyl and authentic alpha-MSH.

Animals↗
PubMed1986

Hyperbaric oxygen in the treatment of radiation-induced optic neuropathy.

Four patients with radiation-induced optic neuropathies were treated with hyperbaric oxygen. They had received radiation therapy for treatment of pituitary tumors, reticulum cell sarcoma, and meningioma. Two presented with amaurosis fugax before the onset of unilateral visual loss and began hyperbaria within 72 hours after development of unilateral optic neuropathy. Both had return of visual function to baseline levels. The others initiated treatment two to six weeks after visual loss occurred in the second eye and had no significant improvement of vision. Treatment consisted of daily administration of 100% oxygen under 2.8 atmospheres of pressure for 14-28 days. There were no medical complications of hyperbaria. While hyperbaric oxygen is effective in the treatment of radiation-induced optic neuropathy, it must be instituted within several days of deterioration in vision for restoration of baseline function.

Adult↗
PubMed1986

Influence of antioxidant enzymes in reduction of optic disc edema in experimental optic neuritis.

Acute experimental allergic optic neuritis was induced in 18 adult strain-13 guinea pigs. The animals were divided into three groups. The first group of six animals received intraperitoneal injections of saline, the second group received superoxide dismutase (SOD), and the third group received catalase, given daily, starting 1 week after encephalogenic myelin sensitization. Morphometric analysis of longitudinal histopathologic sections of the optic nerve head revealed that the animals treated with the antioxidant enzyme catalase had a statistically significant reduction of optic disc edema when compared to the saline-treated group. Although animals treated with SOD had less optic disc swelling than the saline-treated controls, the differences were not statistically significant. These results indicate that detoxification of hydrogen peroxide, released by invading inflammatory cells, can reduce the severity of axonal swelling within the optic nerve head.

Animals↗
PubMed1986

[Alpha-melanotropin in the frog brain: regional distribution, quantification and subcellular distribution].

The distribution of alpha-MSH containing neurons was studied by immunofluorescence in the brain of the frog Rana ridibunda. Most immunoreactive cell bodies were found in the ventral hypothalamic area. A rich network of fluorescent fibers was observed in the ventral infundibular region, coursing towards the preoptic area and the ventral telencephalon. Some fibers, directed backwards, project into median eminence. By means of a specific radioimmunoassay, the concentrations of alpha-MSH immunoreactive material has been determined in 10 different regions of the brain. The highest concentrations were observed in the infundibular and the preoptic regions. Using the immunogold technique, electron microscopy showed that immunostaining was restricted to 70-100 nm dense core vesicles in positive cell bodies and fibers. These results suggest that, in addition to well known hormonal (melanotropic) activity, alpha-MSH could play the role of a neurotransmitter in the frog brain.

Animals↗
PubMed1986

[Localization and identification of neuropeptide Y (NPY) in the brain of an anuran amphibian].

A neuropeptide Y (NPY)-like peptide has been localized in the frog brain by the direct immunofluorescence method. Subcellular localization of NPY has been studied by PAP immunocytochemistry and the immunogold technique. Using a specific radioimmunoassay technique, NPY-like peptide has been quantified in various brain areas and the immunoreactive material was characterized by Sephadex G-50 gel filtration and high performance liquid chromatography. The results indicate that frog NPY is closely related to the porcine molecule and largely distributed in the frog brain. This peptide may play both neurotransmitter and neuroendocrine functions in amphibia.

Animals↗
PubMed1985

Immunocytochemical localization of somatostatin-28 in the rat hypothalamus.

Somatostatin-28 (SS28) is considered as a precursor of somatostatin-14 (SS14) and also of the remaining NH2 terminus of SS28, the dodecapeptide SS28 which has been recently characterized. In order to study the cellular and subcellular localization of SS28 in the rat hypothalamus, we have conducted a light and electron microscopic immunocytochemical study, using both the peroxidase-antiperoxidase and the immunogold technique. Several antisera which selectively recognize one or more of these 3 somatostatin-related peptides were used. In serial paraffin sections, it was observed that SS28 was contained in the same neuronal cell bodies which also contain SS28. These neurons were exclusively located in the periventricular nucleus. All the antisera used also produced a strong staining in the external zone of the median eminence. At the electron microscopic level, the 3 peptides were exclusively localized in all the dense core vesicles in cell bodies and axons in the periventricular nucleus and terminals in the median eminence. These results strongly suggest that SS28 and SS14 originate from the precursor SS28 and that processing of the precursor occurs in the cell body. They also support the hypothesis that the 3 peptides are released simultaneously under appropriate stimulation.

Animals↗
PubMed1985

Superior division paresis of the oculomotor nerve.

Five patients with isolated ptosis and a paresis of ocular elevation in abduction consistent with an isolated superior division III nerve palsy are reported. In all instances the III nerve appeared to be involved before its reported anatomic bifurcation into a superior and an inferior division, in the anterior cavernous sinus.

Adult↗
PubMed1985

Immunohistochemical identification of the uncoupling protein in rat brown adipose tissue.

Brown adipose tissue mitochondria are characterized by the presence of an uncoupling protein that gives them an exceptional capacity for substrate-controlled respiration and thermogenesis. The specific localization of this protein in rat brown adipocytes was demonstrated using an immunohistochemical technique, the peroxidase-antiperoxidase (PAP) method. Light microscopy observations showed that serum antibodies raised against the uncoupling protein selectively reacted with multilocular brown adipocytes. No labeling could be detected in either unilocular adipocytes, capillaries, or muscle fibers (striated and vascular smooth muscle). Staining was more intensive in certain adipocytes than in others, suggesting the presence of cellular heterogeneity. The specificity of the staining technique was demonstrated by showing that treatment of the preparations with antiserum saturated with an excess of uncoupling protein almost entirely inhibited brown adipocyte labeling. The specificity and selectivity of the PAP method allow the clear differentiation of uncoupling protein-containing adipocytes from other cellular types, suggesting that this immunohistochemical technique will represent an extremely useful tool for studying adipocyte function and differentiation.

Adipose Tissue, Brown↗
PubMed1985

[Intermediate lobe of the amphibian pituitary gland: an endocrine gland with multiple secretions and under multi-hormonal control].

The cells of the frog pars intermedia synthesize a 36 000 (36K) protein called proopiomelanocortin (POMC). After [3H]glucosamine incorporation, separation of newly synthesized products by SDS-polyacrylamide gel electrophoresis showed that this 36K protein was glycosylated. Tryptic mapping revealed only one site of glycosylation and showed that the carbohydrate side-chain was located in the N-terminal region of POMC. The 36K protein was not released by the melanotrophs, but it generated, through specific intracellular proteolytic cleavage, a number of smaller peptides which were subsequently released. These peptides were identified by various methods including selective amino-acid incorporation, HPLC purification, acid-urea gel electrophoresis, tryptic and chymotryptic mapping, assay of melanotropic activity, radioimmunoassays and immunoprecipitations. Some of the newly synthesized N-terminal (18K) fragment of the POMC was secreted intact while a portion of it was further processed, via an intermediate peptide, to give mature gamma-MSH. All three of these peptides were glycosylated. In addition, the mature peptide (gamma-MSH) exhibited a low but significant melanotropic activity. The C-terminal portion of the prohormone was very rapidly processed to give des N alpha-acetyl alpha-MSH, corticotropin-like-intermediate lobe peptide (CLIP) and beta-endorphin. Authentic alpha-MSH was always absent in cellular extracts: acetylation to give rise to alpha-MSH was a late enzymatic process strictly linked to hormonal release. Since acetylation of alpha-MSH is required for full biological activity of this peptide, it is possible to conceive that this later step could be under neuroendocrine control. Using the perifusion technique we have been able to show the complexity of the control mechanisms regulating amphibian melanotrophs. It is generally accepted that the aminergic innervation of the intermediate lobe of the pituitary is involved in the hypothalamic control of melanotropin release. We have demonstrated that, in amphibians, dopamine inhibits alpha-MSH secretion through D2-type dopaminergic receptors whereas norepinephrine and (or) epinephrine stimulate alpha-MSH secretion via beta-adrenergic receptors. The existence of peptidergic fibers within parenchymal cells of the pars intermedia has been demonstrated. Evidence for TRH-containing fibers has been obtained by immunohistochemistry. Using a specific radioimmunoassay for TRH, we have confirmed the presence of TRH in the neurointermediate lobe of the frog. We have shown that TRH is a powerful MSH-releasing factor in these animals.(ABSTRACT TRUNCATED AT 400 WORDS)

Amphibians↗
PubMed1984

Acute and chronic experimental optic neuritis. Alteration in the blood-optic nerve barrier.

Horseradish peroxidase (HRP) tracer studies in experimental allergic optic neuritis showed alterations in the blood-optic nerve barrier. Light and electron microscopic examination of the optic disc disclosed intense leakage of HRP at the lamina scleralis, with extracellular staining of both axonal and glial columns, despite the lack of inflammation at this region. The intraorbital optic nerve showed leakage of HRP at the sites of both acute and chronic demyelination. Ultrastructurally, leakage of HRP was confined to the perivascular and extracellular spaces. The mechanism of leakage seems to be by diffusion through the endothelial cell junctions or vesicular transport across the endothelial cells. The high predilection for involvement of the optic nerve in chronic demyelinating disorders may be due to the inherent weakness of the blood-brain barrier at the optic nerve head.

Animals↗
PubMed1984

Electron microscope immunocytochemical localization of neuropeptide Y (NPY) in the rat brain.

In order to identify the neuronal structures as well as the intracellular organelles which contain the newly discovered brain peptide, neuropeptide Y (NPY), we have localized this peptide by immunoelectron microscopy in brain areas containing high levels of NPY. Ultrastructural studies using both the pre- and post-embedding immunoperoxidase techniques have shown that NPY-like immunoreactivity could be observed in neuronal cell bodies, dendrites and axonal processes. In terminals, the reaction product was associated with the large dense core vesicles. Only a small percentage of the positive terminals were seen making synaptic contacts. These results support the hypothesis that NPY can act as a neurotransmitter or neuromodulator.

Animals↗
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↗