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Biomedical subjects

G Pelletier

Publications and source records attributed to G Pelletier.

At least 415 records · Page 23Linked to original sources

Serotonin innervation of neuropeptide Y-containing neurons in the rat arcuate nucleus.

Structural non-synaptic appositions between serotonin (5-HT) nerve endings and neuropeptide Y (NPY)-containing neurons were demonstrated in the rat arcuate nucleus by means of a combined radioautographic and immunocytochemical detection of [3H]5-HT uptake sites and NPY-immunoreactivity. Such cellular relationships are proposed to constitute a morphological substrate for putative 5-HT/NPY interactions in neuroendocrine hypothalamus.

Animals↗

Immunocytochemical distribution of neurons containing a peptide derived from thyrotropin-releasing hormone precursor in the rat brain.

Using an antiserum to a 22-amino acid synthetic peptide corresponding to sequence 178-199 of the precursor of thyrotropin-releasing hormone (pro-TRH), we have studied by immunocytochemistry the distribution of immunoreactive pro-TRH in the rat brain. Immunostaining found in several regions of the brain had a distribution similar to that previously reported for TRH. Reaction product was found in both neuronal cell bodies and processes. These results indicate that peptide(s) derived from pro-TRH are transported along the axons with TRH itself. Moreover, the presence of immunoreactive nerve endings in different areas including the median eminence suggests that this material might have a neuromodulator/neurotransmitter role in the central nervous system and could also be released into the pituitary portal plexus.

Animals↗

Prognostic value of HBsAg/IgM complexes in hepatitis B patients: nature of the proteins involved.

Hepatitis B surface antigen (HBsAg)/IgM complexes were measured using a solid phase radioimmunoassay in sera of patients with acute or chronic hepatitis B. These complexes were found in 6 out of 18 patients four weeks after the onset of the disease and only one of them developed chronic hepatitis. HBsAg/IgM complexes correlated with HBsAg, hepatitis B e antigen (HBeAg), and pre-S2 concentration. The precipitation of HBsAg/IgM reactivity by polyethylene-glycol (PEG) and the binding of this activity to the surface of certain uncoated enzyme linked immunosorbent assay (ELISA) plates indicates that HBsAg/IgM positivity may reflect the presence of circulating complexes in serum. HBsAg and pre-S2 were found as components of the complexes but anti-HBs, anti-pre-S2, and polymerized human serum albumin (pHSA) were not. An immune binding between HBsAg and IgM is still questionable. Whatever the nature of the HBsAg/IgM complexes, their detection does not seem to be an earlier indicator of prognosis than HBeAg and/or pre-S2.

Adult↗

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↗

Immunohistological demonstration of a substance related to neuropeptide Y and FMRFamide in the cephalic and thoracic nervous systems of the locust Locusta migratoria.

A neuropeptide related to the mammalian neuropeptide Y (NPY) is present in various neurosecretory cells (NSC) of the cephalic and thoracic nervous systems of the insect Locusta migratoria. Immunoreactive perikarya are detected in the protocerebrum, tritocerebrum, optic lobes and the suboesophageal and thoracic ganglia. They give rise to many immunoreactive processes that ramify extensively throughout the neuropiles. In the brain, prominent axon bundles tightly surround the tractus I to the corpora cardiaca. This fiber pattern suggests that the NPY-like substance may have a neuromodulator and/or neurotransmitter function. This substance may also have a neurohormonal role, since some immunoreactive tracts penetrate into neurohaemal organs via the nervi corporis cardiaci II and the thoracic median nerves. NCS containing NPY-like neuropeptide also display an FMRFamide-like immunoreactivity (except for the abdominal part of the metathoracic ganglion). NPY or FMRFamide antisera are not inactivated after preabsorption with FMRFamide or NPY, respectively. It might therefore be inferred that in locust NSC these two antisera recognize two distinct antigenic sites belonging either to a large polypeptide, or to two distinct neuropeptides.

Amino Acid Sequence↗

Use of slide-mounted frozen sections to study the autoradiographic localization, characterization and quantification of luteinizing hormone-releasing hormone receptors in the rat pituitary.

Slide-mounted frozen sections were used for the autoradiographic localization, characterization and quantification of LHRH receptors. The radioligand was the iodinated LHRH agonist [D-Ser(TBU)6, des-Gly-NH2(10)]LHRH ethylamide (LHRH-A). Specificity studies demonstrated that specific labeling was concentrated in the anterior pituitary lobe and that the tracer could be displaced with either LHRH or LHRH-A. Scatchard analysis of saturation curves gave one-site interactions with a high affinity (Kd = 0.48 +/- 0.1 nM) for the iodinated LHRH-A. In order to evaluate the effects of different treatments on the LHRH receptor concentration, the effects of sex steroids on the density of radioautographic reaction were evaluated using an image analyzer. The results obtained were similar to those previously reported with classical binding assays. Thus, this simple technique appears suitable for the study of localization and characterization of LHRH receptors in different experimental conditions.

Animals↗

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↗

Anti-idiotypic antibodies as probes for the determination of idiotopes shared by human and monoclonal murine antibodies.

Murine monoclonal antibodies (MAbs) against rye grass Group I (rye I) allergen were previously produced (290A-167, 348A-6 and 539A-6) and are further characterized herein. By competitive binding to polystyrene-bound allergen, it was shown that the three MAbs are directed against three different non-overlapping epitopes on rye I. Human rye I-specific IgG isolated from the serum of one rye grass sensitive patient could recognize the same or closely related epitopes than those recognized by MAbs. Antisera were then raised in rabbits against purified F(ab')2 fragments of human rye I-specific IgG and F(ab')2 fragments of 539A-6 MAb. The antisera were rendered idiotype specific by adsorption with insolubilized non-specific human IgG from the same donor, insolubilized normal murine immunoglobulins and finally with rye I. As shown by competitive binding to polystyrene-bound allergen, rabbit anti-idiotypic antibodies (anti-ID antibodies) produced against F(ab')2 fragments of human rye I specific IgG could inhibit the reaction between two MAbs (290A-167 and 539A-6) and the relevant allergen. Furthermore, human rye I specific IgG could inhibit the binding of 125I-labelled 539A-6 MAb to its specific anti-ID antibody to a significant degree. Human autoanti-idiotypic antibodies were also shown to inhibit the reaction between the three anti-rye I MAbs and the antigen. These observations suggest that cross-reactivity exists between idiotypic determinants of human rye I-specific IgG and the three murine MAbs, which implies structural similarity in the V genes coding for the variable region of the antibody from these two different species.

Allergens↗

Studies on the physiological role and mechanism of action of neuropeptide Y in the regulation of luteinizing hormone secretion in the rat.

It has been recently shown that intraventricular or systemic injection of neuropeptide Y (NPY) can produce a decrease in plasma luteinizing hormone (LH) levels in castrated rats of both sexes. In order to evaluate the physiological role of NPY in the regulation of LH secretion in the female rat, we proceeded to immunoneutralization experiments using specific antibodies to NPY. Injection of 0.5 ml antiserum to NPY produce a 20-fold increase of LH plasma levels, whereas injection of preimmune serum did not modify the plasma concentrations of LH. To investigate the possibility that catecholamines or serotonin might be involved in the effect of NPY in LH secretion, castrated rats were treated with alpha-methylparatyrosine (alpha-MPT), an inhibitor of catecholamine biosynthesis, or received an i.c.v. injection of the neurotoxin 5-7-dihydroxytryptamine (5,7-DHT) prior to the intraventricular injections of NPY. The pretreatment with alpha-MPT could not prevent the decrease of plasma LH induced by NPY injection whereas the pretreatment with 5,7-DHT reversed the effect of NPY injection. The anatomical connection between LH-releasing hormone (LHRH) and NPY neuronal systems were also investigated using double immunostaining technique. It appeared that NPY endings are in apposition to LHRH cell bodies in the preoptic area in proximity to the organum vasculosum of the lamina terminalis (OVLT).(ABSTRACT TRUNCATED AT 250 WORDS)

5,7-Dihydroxytryptamine↗

Distribution of estrogen receptors in the rat pituitary as studied by in situ hybridization.

The localization of estrogen receptors in the rat pituitary gland was performed by in situ hybridization. The probe used was a synthetic oligonucleotide probe labelled with 35S complementary to the mRNA coding for a fragment (1-24) of estrogen receptor. Strong labelling was observed in cells of the anterior and intermediate lobes whereas the posterior lobe remains unlabelled. These results suggest that not only the anterior lobe but also the intermediate lobe is a target for estrogens.

Animals↗

Autoradiographic localization of beta-adrenergic receptors in rat oviduct.

Catecholamines are known to have an inhibitory effect on oviductal smooth musculature by changing the adrenergic receptors activity. In order to further investigate the role of epinephrine and/or norepinephrine in oviduct, the distribution of beta-adrenergic receptors has been studied in the rat oviduct, using in vitro autoradiography and [125I]cyanopindolol (CYP), as radioligand. The specificity of the labelling and the characterization of receptor subtypes in different cell types was achieved by displacement of radioligand with increasing concentrations of zinterol, a beta-adrenergic agonist with preferential affinity for the beta 2-adrenoreceptor subtype and practolol, a beta-adrenergic antagonist that binds preferentially to beta 1-subtype. Quantitative estimation of ligand binding was achieved by densitometry. It was shown that the vast majority of beta-receptors were of the beta 2-subtype and were found in smooth muscle layers as well as in the epithelium. The latter localization suggests a role for epinephrine and/or norepinephrine on the oviductal epithelium.

Animals↗

High resolution autoradiographic localization of LHRH receptors in the rat anterior pituitary using fixed sections.

The cellular and subcellular localization of luteinizing hormone-releasing hormone (LHRH) receptors in rat anterior pituitary has been studied at the light and electron microscopic levels using in vitro autoradiography performed on fixed pituitary sections. The iodinated LHRH agonist [D-Ser(TBU)6,des-Gly-NH10(2)]LHRH ethylamide was used as the ligand. In semithin sections of pituitaries from castrated rats, specific labeling was exclusively localized over the castration cells. At the ultrastructural level, the analysis of silver grain distribution performed in gonadotrophs revealed that the plasma membrane, rough endoplasmic reticulum, and Golgi apparatus were significantly labeled. These findings are consistent with previous reports indicating the presence of specific LHRH binding sites in the plasma membrane. This is also the first demonstration that LHRH receptors are associated with intracytoplasmic organelles. Moreover, this ultrastructural technique should be very useful to localize with precision LHRH receptors in other organs, especially the brain.

Animals↗

Serum growth hormone release during a 60-hour period in growing pigs.

Growth hormone (GH) profiles were measured during a 60-hour period in four castrated male and five female, 20-week-old Yorkshire pigs. During this period, GH release was not affected by feeding when measured at 2, 4 and 8 hr before and after feeding time. A photoperiod of 12 hr light and 12 hr darkness produced a decrease (P less than .05) in baseline mean GH levels from 4.0 to 3.5 ng/ml during periods of darkness. This effect was observed in both genders. Females and castrated males exhibited (P greater than .05) similar baseline GH levels and identical numbers of GH peaks during feeding and photoperiod studies. However, during these periods, the amplitude of the GH peaks and areas under the GH curves were greater (P less than .05) in females. These results indicate that: 1) feeding did not influence GH secretion; 2) darkness produced a decrease in the baseline GH levels in both sexes; and 3) females secreted more GH than castrated males of the same age.

Animals↗

Lymphocyte subpopulation monitoring in cyclosporine-treated patients following heart transplantation.

Circulating lymphocyte subpopulations were studied in 18 consecutive patients treated with cyclosporine-prednisone immunosuppression during the first month following heart transplantation. Eleven patients showed no evidence of graft rejection. There were eight episodes of acute rejection demonstrated at endomyocardial biopsy in 7 patients. Three patients were treated with corticosteroids, 3 were treated with rabbit antithymocyte globulin (RATG), and 1 died before treatment (early mortality: 5.6%). Using the monoclonal antibody technique, 150 determinations of lymphocyte subpopulations were performed and were correlated with 72 endomyocardial biopsy specimens. Cyclosporine immunosuppression caused a significant (p less than 0.05) decrease in total lymphocyte count (38%) and in the number of OKT3 (52%) and OKT4 cells (55%). During acute rejection, total lymphocytes and OKT3, OKT4, and OKT8 cells all increased significantly, but the T4 to T8 ratio did not change significantly. Treatment of rejection with corticosteroids resulted in a moderate but not significant decrease in all T-cell types, whereas RATG caused a marked but not selective decrease in all T-cell groups. In conclusion, T cells decrease with cyclosporine immunosuppression and with treatment of rejection and increase at onset of rejection, but the T4 to T8 ratio has no predictive value for the diagnosis and severity of rejection, and the sensitivity of the method does not permit its use to assess the degree of immunosuppression with cyclosporine following heart transplantation.

Adolescent↗

Mixed growth hormone and prolactin-secreting human pituitary adenomas: a pathologic, immunocytochemical, ultrastructural, and immunoelectron microscopic study.

A study of 30 adenomas from patients with signs of growth hormone (GH) and prolactin (PRL) hypersecretion revealed the presence of mammosomatotroph cells (MSC) containing both hormones in all cases. Although the number of immunostained cells varied from case to case, in 14 of 25 tumors, all stained cells were MSC. Nine tumors had the ultrastructural appearance of densely granulated growth hormone adenomas, while 11 cases resembled sparsely granulated growth hormone adenomas with frequent fibrous bodies. Exocytosis was present in six of these 11 cases, a feature unusual for pure growth hormone adenomas. Nine tumors consisted of a mixture of cells with the morphology of GH and PRL cells. In the four cases examined, immunoelectron microscopy using double immunolabeling with protein A-gold particles revealed the presence of secretory granules containing both hormones in some tumor cells recognized as mammosomatotroph cells.

Adenoma↗

The luteinizing hormone-releasing hormone (LHRH) agonist, [D-Trp6, des-Gly-NH2(10)]-LHRH ethylamide, has no direct effect on spermatogenesis in the rat.

Treatment of intact rats with luteinizing hormone-releasing hormone (LHRH) agonists has been shown to produce atrophy of a variable number of testicular seminiferous tubules. These findings raised the question of a possible direct versus indirect action of LHRH agonists on spermatogenesis. To answer this question, we treated hypophysectomized rats with the LHRH agonist [D-Trp6, des-Gly-NH2(10)]-LHRH ethylamide, dihydrotestosterone (DHT), or a combination of these two compounds for a period of 1 mo. Treatment of hypophysectomized animals with the LHRH agonist alone had no significant effect on the atrophy of seminiferous tubules found after hypophysectomy. DHT, however, maintained spermatogenesis at 80% of the level seen in intact animals. When DHT and the LHRH agonist were administered in combination, the stimulatory effects of DHT were observed with no significant interference caused by the LHRH agonist. This study shows that an LHRH agonist has no direct effect on the morphology of the seminiferous tubules in the absence of the pituitary gland and strongly suggests that the atrophy observed in the testis after LHRH agonist treatment in intact animals is mediated by the LHRH agonist-induced changes in luteinizing hormone secretion and/or direct action of the peptide on Leydig cells.

Animals↗