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Biomedical subjects

G Morel

Publications and source records attributed to G Morel.

At least 91 records · Page 5Linked to original sources

Rat granulosa cells express the gonadotrophin-releasing hormone gene: evidence from in-situ hybridization histochemistry.

There is still debate as to whether natural sequence gonadotrophin-releasing hormone (GnRH) is produced in the mammalian gonads and concerning its potential role as a paracrine modulator of gonadal function. To address this question, we have used in-situ hybridization histochemistry with an oligonucleotide probe complementary to the GnRH decapeptide coding sequence, to determine the cellular site(s) of expression of the GnRH gene in rodent ovaries. GnRH mRNA was detected in granulosa and thecal cells from ovarian follicles at all stages of development (primary-->Graafian), with no significant change in grain density during follicular development. The granulosa cell compartment always contained more mRNA than the thecal cell compartment. Corpora lutea expressed the GnRH gene to the same extent as thecal cells. These results indicate that preproGnRH mRNA is detectable under physiological conditions in the mammalian ovary, though whether this produces authentic GnRH decapeptide or an alternative protein product is not known. The physiological significance of these findings remains to be determined.

Animals↗

Light and electron microscopic immunocytochemical evidence that delta sleep-inducing peptide and gonadotropin-releasing hormone are coexpressed in the same nerve structures in the guinea pig median eminence.

The relationships between delta sleep-inducing peptide (DSIP) and GnRH immunoreactivity within the guinea pig median eminence are investigated by light and electron microscopic immunocytochemistry. Indirect immunofluorescence and elution-restaining experiments show that at the light microscopic level the distribution patterns of DSIP and GnRH immunoreactivity are indistinguishable. This suggested the possible coexistence of both immunoreactivities within the same fibers and neurosecretory endings. At the electron microscopic level, a preembedding double-immunolabeling technique using both indirect immunoperoxidase and immunogold methods, clearly indicate that DSIP and GnRH immunoreactivity are frequently colocalized within single secretory granules. In addition DSIP/GnRH immunoreactive nerve endings were also observed often in close proximity to tanycyte elements. Taken together, the present results provide for the first time ultrastructural evidence for the presence of DSIP immunoreactivity and demonstrate that DSIP and GnRH immunoreactivities may be coexpressed within the same neuronal elements in the median eminence.

Animals↗

Arginine-vasopressin in anterior pituitary cells: in situ hybridization of mRNA and ultrastructural localization of immunoreactivity.

The hypothalamic nonapeptide arginine-vasopressin (AVP) exerts several distinct receptor-mediated actions on pituitary cells. Although hypothalamic AVP reaches the anterior pituitary via well-defined pathways, there is now accumulating evidence that AVP may also be produced endogenously in anterior pituitary cells. Using in situ hybridization, we demonstrate here the presence of AVP mRNA in the anterior pituitary of the rat. The observed grain density over pituitary cells was, however, greater than 10-fold lower than the one observed over AVP producing neurons present in the supraoptic and paraventricular nuclei of the hypothalamus. Immunoelectron microscopic analysis using two different AVP-specific antibodies revealed that the distribution of AVP-like immunoreactivity (AVP-LI) in the anterior pituitary is cell-specific. AVP-LI is most abundant in corticotrophs, followed by lactotrophs, gonadotrophs and thyrotrophs. On the other hand, there is complete absence of AVP-LI from somatotrophs. Interestingly, all pituitary cells in which AVP-LI is detected also represent potential target sites for AVP action. A minor fraction of AVP-LI was found to be membrane-associated and may originate, at least in part, from extrapituitary sources. This fraction likely represents receptor-bound peptide. The bulk of AVP-LI, however, was present in the cellular cytoplasm, not associated with any specific ultracellular structure. Specifically in corticotrophs, AVP-LI was excluded from secretory granules. However, our finding of AVP mRNA in anterior pituitary cells indicates that intracellular AVP-LI includes endogenously produced peptide, suggesting a paracrine and/or autocrine action.

Animals↗

Uptake and ultrastructural localization of a [125I] growth hormone releasing factor agonist in male rat pituitary gland: evidence for internalization.

GRF was isolated from a human tumor of the pancreas and characterized. GRF stimulates the in vivo and in vitro secretion of GH. The present study was designed to find out whether human (h) GRF agonist could be internalized and to determine the subcellular localization of internalized peptide in somatotrophs. Autoradiography was performed on rat anterior pituitary glands removed at specific time intervals (2-60 min) after iv injection of monoradioiodinated [125I] (His1,Nle27) hGRF (1-32) NH2. Administration of an excess of unlabeled hGRF agonist along with the radioiodinated hormone prevented the uptake, indicating the specificity of the reaction. At the ultrastructural level only the somatotrophs appeared to contain silver grains. The main effect of hGRF agonist injection on the cytological aspect of the somatotrophs was a decrease in the area occupied by secretory granules, accompanied inversely, by an increase in that of the Golgi complex. The time course study in somatotrophs showed that five compartments (plasma membrane, secretory granules, cytoplasmic matrix, nuclear membrane, and lysosomes) have distinct marked labeling patterns. Plasma membrane, secretory granules, and nuclear membrane were labeled throughout the time course studied (2-60 min after injection). Cytoplasmic matrix was labeled 5 min post injection and lysosomes 15 and 30 min after injection. The Golgi complex, mitochondria, rough endoplasmic reticulum, and nucleus matrix were not labeled. The findings show the cellular specificity of GRF uptake by somatotrophs and the internalization process from the plasma membrane to the intracellular organelles (secretory granules, lysosomes, and nuclear membrane). Labeling of the secretory granule compartment suggests that granules may bind and protect internalized peptide from lysosomal degradation. The appearance of label on the nuclear membrane suggests that GRF may have effects on the translocation of messenger RNA from nucleus to cytoplasm in somatotrophs.

Animals↗

[A focus of multiple sclerosis in Bretagne: HLA markers and evaluation of consanguinity].

In a previous study, we established the overall prevalence of multiple sclerosis at 25 per 100,000 inhabitants in the French province of Brittany and found that the geographical distribution was uneven with four circumscribed high prevalence areas with more than 45 per 100,000. We conducted the present study to try to ascertain whether the existence of such clusters of MS could be explained by genetic factors, using two ways: the major histocompatibility markers and the frequency of intermarriage. Among the four areas of high prevalence, we examined the one with the highest prevalence, exempt from migratory movements over the last 100 years. We compared this studied population to a sample of 1005 healthy unrelated individuals coming from all over Brittany, designated as general Brittany population. A large sampling (about 25%) of the population accepted to take part in the study. Regarding HLA markers, we observed in the high prevalence area an increases percentage of B7, B8, B12, DR4 and a decreased percentage of B5, compared with the Brittany control population. The consanguinity coefficient in the population of high MS prevalence area was steadily much higher than the one in neighbouring areas with a low MS prevalence during a long period from 1896 to 1945. Under the reserve of non-randomized sampling of individuals for HLA marker study, our findings yielded a double argument in favor of genetic factors influencing the presence of clusters of MS cases: the particularities of HLA phenotype pattern and the increased consanguinity coefficient. The particularities of HLA polymorphism observed in the high prevalence area are likely to be the consequence of the intermarriage habits of this population.

Adult↗

[Hydroxylation of debrisoquin in Parkinson's disease].

The accumulation of environmental substances, due to deficiency of liver detoxication processes, is one of the hypotheses put forward in the search of the cause of Parkinson's disease. The debrisoquine test was used to evaluate the capacity for hydroxylation of twenty patients treated for Parkinson's disease. No difference was found with the results obtained in non Parkinsonians: other metabolic pathways, notably S-methylation, should be explored.

Adult↗

Ultrastructural distribution of somatostatin-14 and -28 in rat adrenal cells.

Previous studies have shown that somatostatin modulates angiotensin-induced aldosterone secretion by adrenal glomerulosa cells. This effect is mediated through specific receptors which do not show any preference for somatostatin-14 (S14) or the N-extended form somatostatin-28 (S28). The study of the distribution of 125I-Tyr [Tyr0, DTrp8] S14- and 125I-Tyr [Leu8, DTrp22, Tyr25] S28-binding in frozen sections of the rat adrenal by autoradiography indicated that both peptides bind to similar loci. High concentrations of binding sites were observed in the zona glomerulosa, and low concentrations were detected in the medulla. At the ultrastructural level, immunocytochemistry after cryoultramicrotomy revealed endogenous S14- and S28-like immunoreactive material in zona glomerulosa and in medulla. In glomerulosa cells, immunoreactive material was localized at the plasma membrane level, in the cytoplasmic matrix, in the mitochondria, and in the nucleus. S14- and S28-like materials were detected in both epinephrine and norepinephrine-storing cells of the adrenal medulla. In these cells, the distribution of either immunoreactive product was similar; it was observed in cytoplasmic matrix, secretory granules and nucleus, but not at the plasma membrane level. In situ hybridization does not reveal somatostatin mRNA in zona glomerulosa or medulla. These results demonstrate that S14 and S28 bind to, and are taken up by zona glomerulosa and adrenal medullary cells, but are not produced by these cells.

Adrenal Glands↗

Localization of endogenous osteocalcin in neonatal rat bone and its absence in articular cartilage: effect of warfarin treatment.

Immunocytochemistry after cryoultramicrotomy was used to localize endogenous osteocalcin in bone (calvaria, femoral diaphysis) and epiphyseal femoral cartilage from 8-day-old rats treated (or mot) for 7 days with warfarin. Ultrathin frozen sections were incubated with goat antiserum against rat osteocalcin at high dilutions (2 x 10(-4) to 2 x 10(-6). In calvaria and femur of untreated rats, endogenous osteocalcin was observed in osteoblasts (cytoplasm and nucleus) and in the collagenous matrix. Osteocalcin appeared progressively in osteoblasts and bone matrix in the mineralization front, then increased in the regions of extended calcification. Osteocalcin was also detected in osteocytes but was not as abundant as in osteoblasts. In bone samples of warfarin-treated rats, endogenous osteocalcin was only detected in bone matrix but not in osteoblasts. Furthermore, osteocalcin was only observed if antiserum was not very dilute (2 x 10(-2). In cartilage (hypertrophied and degenerative zones), osteocalcin was not observed in matrix and chondrocytes. However, it was found in the vicinity of matrix vesicles at the initial loci of calcification. Osteocalcin was never detected in the cartilage of warfarin-treated rats. Our results provide ultrastructural immunocytological evidence for the localization of endogenous osteocalcin in osteoblasts, the presence of osteocalcin in bone matrix and a direct gradient between the presence of osteocalcin and the calcification process. Osteocalcin is absent from cartilage, except possibly close to calcifying matrix vesicles. Warfarin inhibits the formation of osteocalcin.

Animals↗

Human immunodeficiency virus type 1 infection of U937 cells promotes cell differentiation and a new pathway of viral assembly.

The differentiation of U937 monoblastoid cells after human immunodeficiency virus type 1 (HIV-1) infection was studied using the following approaches: reverse transcriptase activity measurement, immunofluorescence labeling, and electron microscopy. For comparison, uninfected U937 cells were induced to differentiate from monocyte to macrophage by phorbol 12-myristate 13-acetate (PMA) or retinoic acid (RA) treatment. Both infected and drug-treated cells showed important and similar ultrastructural cell modifications, with a phenotype that decreased in monocyte specificity and increased in that of macrophages. When U937 cells were induced to differentiate upon HIV-1 infection, a very different pathway of viral production was observed. Production and accumulation of the virus in a vacuolar compartment of intracytoplasmic origin and escape to the antiviral lysosomal activity could explain virus persistence. This makes the cell system a good model with which to study the relationship between HIV-1 production and cell differentiation.

Antigens, CD↗

Assessment of urinary rat beta 2-microglobulin by enzyme immunoassay.

An enzyme immunoassay (EIA) was developed using unlabelled and peroxidase-labelled rabbit antibodies to assess urinary rat beta 2-microglobulin (beta 2-m) excretion. It consisted in the adsorption of rabbit anti-rat beta 2-m immunoglobulin to a polystyrene surface, the addition of beta 2-m samples or standard and the addition of peroxidase-labelled rabbit anti-rat beta 2-m immunoglobulin. After addition of hydrogen peroxide and o-phenylenediamine, the enzyme activity of the solid phase was measured photometrically at 490 nm. Analytical recovery of pure beta 2-m in urine was 102%. From determinations made on normal female and male rats, the ranges of 24-h urine beta 2-m individually excreted were found to be 0.84-4.77 and 3.7-17.3 micrograms, respectively. The means +/- SEM were 2.49 +/- 0.17 and 10.2 +/- 0.55 micrograms. EIA was of value in evidencing the tubule-damaging properties of sodium chromate and hexachloro-1,3-butadiene in rats.

Animals↗

Randomized double-blind trial of injectable heptaminol for controlling spontaneous or bromocriptine-induced orthostatic hypotension in parkinsonians.

Heptaminol is a molecule with experimental cardiovascular analeptic properties. In this double-blind vs placebo trial, the potency, so far unproven, of the injectable form of a 626 mg dose of heptaminol chlorhydrate on spontaneous or induced orthostatic hypotension (OH), was assessed. Nineteen patients were included in the study: 7 displayed spontaneous OH, and in the other 12 OH was induced by bromocriptine, as monitored 103 min/after an oral intake of 6.6 mg on average. Neither spontaneous nor induced OH were recorded in 32% of the Parkinsonian population registered, with no obvious distinctive characteristics. Potency tilt-trials, performed 15, 30 and 45 min after parenteral administration of heptaminol, revealed a significant and expressive potency of the molecule on the systolic blood pressure after 15 min (P less than 0.05). Clinical and biological tolerance was excellent. Low plasma renin activity and the absence of response to orthostatism indicated, in this population of Parkinsonian extrapyramidal syndromes, a loss in positive tonus likely to be of sympathetic origin. The anti-hypotensive action of heptaminol does not seem to be related to any renal or even sympathetic interaction.

Blood Pressure↗

[Treatment of the dislocated hip from walking age to 5 years].

Review of 93 previously not-treated congenital dislocations of the hip, followed at the Institute Calot between 1965 and 1985. Three dislocations were treated by immediate surgical reduction and for the remaining 90 cases, the basic treatment was orthopedic reduction followed by innominate osteotomy. Femoral derotation osteotomy was also performed in 13 cases. Out of them 6 with various effect. Surgical reposition was necessary in 9 cases. The main technical aspects and problems are described. The complications were: abnormal development of the superior femur: 4 cases. Recurrent luxation and subluxation: 5 cases. Early fusion of the triradiate cartilage: 13 cases. Mostly transitory stiffness: 17 cases. Superficial sepsis: 7 cases. Greenstick fractures: 5 cases. Forty five hips were reviewed at the end of the growth period. The sphericity of the femoral head was evaluated according to the criteria of Mose and the coverage of the head according to the criteria of Lequesne. The results were presented according to the classification of Séverin, modified by Robert and Seringe with additional criteria of Mose for evaluation of the morphology of the head. Out of 45 cases there were 36 normal hips (80%), with 12 perfect hips.

Child, Preschool↗

Localization of serotonin-like immunoreactivity in the Merkel cells of pig snout skin.

The presence of serotonin in the Merkel cells of pig snout epidermis was investigated by the peroxidase-antiperoxidase immunohistochemical technique. Serotonin-like immunoreactive Merkel cells were found in groups located at the base of epidermal rete pegs and in the external root sheath of sinus hair follicles (vibrissae). Immunoreactivity was stronger on the basal side of the Merkel cells, where dense-cored granules are most numerous. Neither the nerve terminal associated with the Merkel cell nor the neighbouring epidermal cells were immunostained. These results are the first evidence of serotonin-like immunoreactivity in mammalian Merkel cells. The fact that immunoreactivity is strongest in those parts of the Merkel cells with the highest granule density suggests that in these cells serotonin is probably localized in the dense-cored granules.

Animals↗

[Effects of Chiari osteotomy on acetabular development in the growing subject. Experimental study in puppies].

A Chiari osteotomy was performed on 10 dogs aged 2 months in order to assess the risks of mechanical or ischaemic loss of growth potential in the peri-acetabular cartilage. The development of the acetabular roof was studied by radiographs, microradiographs and histological assessment between 2 and 12 months. Apart from a few mechanical complications, the principal findings were the absence of any signs of necrosis, persistence of a strongly osteoblastic roof and readaptation of the trabecular pattern with articular remodelling, provided the mechanical conditions were satisfactory. The osteotomy must be sufficiently low, while remaining extra-articular, and adequate medial shift must be obtained. The interposed capsular tissue can then become dense and fibrous, providing a sliding surface.

Acetabulum↗

Immunocytochemical localization of ACTH-like immunoreactivity in rat adrenal glomerulosa and fasciculata cells.

The role of ACTH in the synthesis of the adrenocortical hormones has been largely described. In order to investigate the localization of this peptide at the subcellular level of the adrenal glomerulosa and fasciculata cells, an immunocytological method was used. Rat adrenals were fixed and frozen. Ultrathin sections obtained by cryoultramicrotomy, were incubated with anti-beta (1-24) ACTH or anti-alpha (17-39) ACTH sera. The antigen-antibody reaction was detected by PAP complexes (revealed by 4-chloro-1-naphthol) or with protein A-colloidal gold or IgG-colloidal gold. The results obtained were the same whatever the antisera of the technique employed. All the cells of the adrenal zona glomerulosa and zona fasciculata were labelled. ACTH-like immunoreactivity in zona glomerulosa and zona fasciculata cells was observed at the plasma membrane level, in cytoplasmic matrix, mitochondria and nucleus (in the euchromatin close to the heterochromatin aggregations and, occasionally, associated with the nucleolus). No immunoreactivity was observed when non-immune serum or anti-ACTH serum preincubated with ACTH were used, nor there was any modification of the immunocytochemical reaction when anti-ACTH serum incubated with heterologous antigens was employed. These data, demonstrate the presence of endogenous ACTH in both adrenal glomerulosa and fasciculata cells, and suggest that the peptide is internalized after binding to the plasma membrane.

Adrenocorticotropic Hormone↗

Presence of angiotensin II in the adult male rat anterior pituitary gland: immunocytochemical study after cryoultramicrotomy.

Angiotensin II (AII)-like immunoreactivity and binding sites have recently been demonstrated at the pituitary level. This peptide also exerts a stimulatory effect on anterior pituitary hormone release. Immunocytochemistry on ultrathin sections obtained by cryoultramicrotomy was used with the aim of localizing endogenous AII-like material at the cellular and subcellular levels of the anterior pituitary gland. AII-like immunostaining was observed only in gonadotrophs, lactotrophs, and corticotrophs. In gonadotrophs, AII-like immunoreactivity was restricted only to secretion granules. In the two other immunoreactive cells, lactotrophs and corticotrophs, immunostaining was observed in the cytoplasm and in the nucleus. In the cytoplasm, AII-like material was visualized in the cytoplasmic matrix and in the secretory granules. In the nucleus, immunostaining was distributed in the euchromatin in the vicinity of the heterochromatin. AII-like immunoreactivity was also seen at the plasma membrane, but only scarcely. No reaction product was found when anti-AII serum preincubated with AII was used. These immunocytochemical results (1) provide evidence that gonadotrophs are only a site of synthesis and/or storage of AII-like material, (2) indicate that lactotrophs and corticotrophs are cells for AII and (3) provide cytological evidence for a direct participation of AII in the regulation of the lactotropic and corticotropic function.

Angiotensin II↗

Immunocytochemical localization, binding, and effects of atrial natriuretic peptide in rat adipocytes.

The metabolic effects of atrial natriuretic peptide (ANP) have not been widely investigated. Since adipocyte cells represent a model system extensively used to examine the metabolic actions of many peptide hormones, we sought to establish whether ANP could bind to adipocyte membranes, alter cyclic nucleotide metabolism, and affect spontaneous or hormone-stimulated lipolysis. Using in vitro autoradiographic techniques, radiolabelled ANP was found to bind specifically to mammary gland fat cells. Additionally, endogenous ANP-like immunoreactivity could be localized in the plasma membrane compartment and cytoplasmic matrix of fat cells, but not in fat vacuoles. [125I]ANP bound to single high affinity sites (Kd = 0.72 nM) in fat cell membranes. The binding was rapid (equilibrium within 1 min at 25 degrees C) and specific. The atrial peptide was capable of stimulating a time- and concentration-dependent increase in cGMP accumulation in isolated adipocytes, but had no effect on spontaneous or stimulated [-)-isoproterenol, ACTH, forskolin) cAMP formation. ANP did not alter the increase in glycerol production stimulated by l-epinephrine in isolated fat cells. While i.v. infusion of ANP stimulated a marked increase in circulating levels of cGMP, the atrial peptide did not alter plasma triglyceride levels. These data demonstrate the presence of specific ANP binding sites on adipocyte membranes and internalization of ANP-associated immunoreactivity. These receptors are biochemically functional given the ability of ANP to augment cGMP formation. The peptide, however, does not exert an action on adipocyte lipolysis. Adipocytes, therefore, represent an ANP target tissue in which the physiological action of the peptide is yet to be defined.

Adipose Tissue↗