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Biomedical subjects

G Morel

Publications and source records attributed to G Morel.

At least 109 records · Page 6Linked to original sources
PubMed1989

Ultrastructural distribution of growth hormone (GH) mRNA and GH intron I sequences in rat pituitary gland: effects of GH releasing factor and somatostatin.

We have measured the distribution of growth hormone (GH) mRNA or intron I sequences by in situ hybridization on ultrathin frozen sections of pituitaries removed from rats injected with saline, GH-releasing factor (GRF) or somatostatin. A 4-fold increase in labeling of the anterior lobe was observed after GRF, no changes with somatostatin. After ultrastructural in situ hybridization, labeling with the GH cDNA probe was specific to somatotrophs. Two populations of cells containing few or many secretory granules were labeled mainly in the cytoplasm or in both cytoplasm and nucleus. Some cells showed labeling at the perinuclear membrane. Hybridization with the GH intron I probe showed the same cell specificity with silver grains mainly located in the nucleus. After GRF, sequences hybridizing to growth hormone cDNA were increased mainly in the nucleus of somatotrophs when compared to mock-injected rats, indirectly suggesting an increase in the transcriptional activity of the growth hormone gene. After somatostatin, the density of labeling in the nucleus was increased suggesting that somatostatin may prevent the export of growth hormone mRNA molecules from the nucleus to the cytoplasm.

Animals↗
PubMed1989

Effect of corticotropin-releasing factor on the release and synthesis of prolactin.

Corticotropin-releasing factor (CRF) has been characterized on the basis of its intrinsic activity to release corticotropin from cultured rat anterior pituitary cells. Injected in intact rats, CRF increases adrenocorticotropic hormone (ACTH) release. Endogenous CRF-like immunoreactivity was detected in the cytoplasm and nucleus of corticotrophs. Using an antirat CRF serum, a similar location of CRF-like immunoreactivity was observed in lactotrophs: cytoplasmic matrix, secretory granules, nucleus and, to a lesser degree, the plasma membrane level were stained. One injection of CRF increased the plasma ACTH concentration 4-fold after 15 min, while plasma prolactin (PRL) increased 2.7-fold 5 min after injection. In vitro, incubation of female pituitary cells with rat CRF (10(-10)-10(-8) M) had no significant effect on PRL secretion. In contrast, after 4 days of in vitro pretreatment with 17 beta-estradiol (10(-9) M), rat CRF stimulated PRL secretion by 42%. In situ hybridization of whole pituitary slices showed that rat CRF injection significantly increased the labeling of corticotrophs using an ACTH-cDNA probe, but had no significant effect on the labeling of lactotrophs using a PRL riboprobe. These results indicate that CRF is a factor which can modulate PRL release but not the synthesis of PRL.

Adrenocorticotropic Hormone↗
PubMed1989

Is atrial natriuretic peptide synthesized and internalized by gonadotrophs?

The present study was designed to determine whether atrial natriuretic peptide (ANP) could be both synthesized and internalized by rat anterior pituitary gonadotrophs. ANP synthesis was assessed by in situ hybridization of ultrathin frozen sections of anterior pituitary to a biotinylated 30-base oligonucleotide to rat ANP mRNA. As revealed by the immunogold technique, only gonadotrophs were labeled by the probe. At the subcellular level, ANP mRNA was observed at both the nuclear and cytoplasmic levels in gonadotrophs, and labeling of the latter compartment was quantitatively more intense. Internalization of ANP was investigated by an in vivo ultrastructural autoradiographic approach. Intravenous injection of [125I]ANP resulted in rapid labeling within 1 min of the plasma membrane, cytoplasmic matrix, secretory vesicle, and mitochondrial compartments and the Golgi apparatus; these compartments were labeled throughout the remainder of the time course studied (1-30 min). Peak labeling of the plasma membrane compartment was at 1 min and diminished from that point; labeling in the Golgi apparatus peaked 5 min postinjection, while in the other compartments labeling was fairly uniform over the time course. The lysosomal compartment was also radiolabeled; however, only 2 and 5 min after injection of [125I]ANP. The findings demonstrate that gonadotrophs can both synthesize and internalize extracellular ANP. These observations can be extended to suggest that ANP has both autocrine and paracrine actions in the anterior pituitary gland. Since the peptide neither stimulates nor antagonizes the release of any anterior pituitary hormone, these actions are probably unrelated to the adenohypophyseal secretory function.

Animals↗
PubMed1989

Evidence for direct action of calcitonin in the rat pituitary gland.

Classic concepts of calcitonin (CT) function have focused on the effects of CT on calcium homeostasis. More recently CT actions on brain and pituitary have been investigated. In order to evaluate the effects of CT on the anterior pituitary gland we studied the action(s) of CT in vitro and visualized endogenous CT in adult male rat pituitary gland by immunocytochemistry on ultrathin sections obtained by cryoultramicomy. In vitro study using dispersed anterior pituitary cells indicated that CT stimulated the secretion of PRL, whereas the secretion of GH, TSH and LH was not affected. CT-like immunoreactivity was observed in lactotropes only. The other pituitary cell types were not immunoreactive. In lactotropes, immunostaining was observed in the cytoplasm and in the nucleus. In the cytoplasm, CT-like immunoreactivity was visuzalized in the cytoplasmic matrix and in the secretory granules. In the nucleus, immunostaining was distributed primarily in the euchromatin, in the vincinity of heterochromatin region. CT-like immunoreactivity was also observed at the plasma membrane but was only scarce. No reaction product was found when anti-CT serum pre-incubated with CT was used. In conclusion, these results bring evidence for a direct action of CT on lactotrope regulation in vitro as well as in intact animals.

Animals↗
PubMed1989

Internalization of atrial natriuretic peptide by adrenal glomerulosa cells.

Internalization of 125I-labelled atrial natriuretic peptide ([ 125I]ANP) by rat adrenal glomerulosa cells in vivo was investigated by means of an ultrastructural autoradiographic approach. One to 30 min after IV injection of [125I]ANP, silver grains were found, at the light microscope level, over all glomerulosa cells; coinjection of 20 micrograms of unlabelled ANP inhibited this binding by 64%. At the electron microscope level, the time-course study indicated maximal silver grain densities in plasma membranes 1 min after IV injection; grains were detected in mitochondria (external membranes and matrix) 2 min after injection, with maximal labelling at 15 min. The cytoplasmic matrix was labelled only 30 min after injection. During the time-course, labelling of nuclei, Golgi apparatus, and lysosomes was minimal. The data suggest that after binding to plasma membranes ANP is rapidly internalized and distributed within glomerulosa cells. The association of radioactivity with mitochondria suggests that ANP may have intracellular sites of action complementary to those on plasma membranes.

Adrenal Cortex↗
PubMed1988

Atrial natriuretic peptide receptors are present in brown adipose tissue.

Binding activities and metabolic effects of atrial natriuretic peptide (ANP) were examined in rat brown fat. ANP binding sites were identified in thin sections of brown adipose. Binding of [125I]-ANP to brown fat membranes was specific, saturable and time-dependent. A single class of high affinity sites (Kd, 1.7 nM; Bmax, 226 fmol/mg protein) was present. ANP increased cGMP synthesis in isolated cells (EC50, 2.5 x 10(-9) M), but did not alter epinephrine-stimulated glycerol production. The results demonstrate the presence of specific ANP receptors in brown fat, activation of which increases cGMP formation; however, since neither ANP nor cGMP affect lipolysis, the biological importance of these receptors remains unknown.

Adipose Tissue, Brown↗
PubMed1988

Somatostatin-28- and somatostatin-14-like immunoreactivities in the rat pituitary gland.

Endogenous SS14- as well as SS28-like immunoreactive materials were detected in both male and female rats by radioimmunoassay and by immunocytochemistry on ultrathin frozen sections. The content of somatostatin-like immunoreactivity was 0.39 +/- 0.08 pg per mg adenohypophysis. Immunoreactivity was localized by immunocytochemistry in three pituitary cell types: somatotrophs, lactotrophs and thyrotrophs, but not in corticotrophs and gonadotrophs. In these three pituitary cell types the SS28- and the SS14-like immunoreactive materials were localized in the cytoplasm and in the nucleus. In the cytoplasm the immunoreactivity was seen in the cytoplasmic matrix and in the secretory granules. In the nucleus it was present mainly in the euchromatin close to the heterochromatin. In somatotrophs and lactotrophs, SS14- and SS28-like immunoreactive materials have been detected at the plasma membrane level. These results suggest that (1) endogenous SS14 and SS28 are present in adenohypophysis in somatotrophs, lactotrophs and thyrotrophs, and (2) the two peptides act on both the cytoplasmic components and the nucleus.

Animals↗
PubMed1988

Synthesis and internalization of atrial natriuretic factor in anterior pituitary cells.

Atrial natriuretic factors (ANF) are a family of peptides originally identified in atrial cardiocytes and having natriuretic, diuretic and vasorelaxatory properties. ANF was recently reported to be synthesized in anterior pituitary cells. In the current study, the cell-specific sites of binding and internalization of ANF in the adenohypophysis, a site of ANF action, were investigated. By in vitro autoradiographic techniques, [125I]ANF was found to bind specifically to both anterior and posterior lobes of the pituitary, but not to the intermediate lobe. 5 min following intravenous injection of [125I]ANF, radiolabel was detected by ultrastructural autoradiography on gonadotrophs, corticotrophs and lactotrophs, but not thyrotrophs or somatotrophs. Label was found at both the plasma membrane level and cytoplasmic matrix, but not in the nuclei, of all three cell types. The cellular and subcellular localization of endogenous ANF-like immunoreactivity was also investigated. Specific immunoreactivity was also detected in gonadotrophs, corticotrophs and some lactotrophs. At the subcellular level in all three cell types, immunoreactivity was localized in the cytoplasmic matrix, on secretory granule membranes, and in mitochondria. Less dense staining was observed in nuclear euchromatin; endoplasmic reticulum and Golgi apparatus were not labelled in any of the cells. The data suggest that notwithstanding their reported ability to synthesize ANF, exogenous ANF can also bind and be internalized by adenohypophyseal gonadotrophs; the binding and presence of endogenous ANF-like immunoreactivity in corticotrophs and lactotrophs suggest that these cells may be sites of action for the atrial peptide.

Animals↗
PubMed1988

ANF and exocrine pancreas: ultrastructural autoradiographic localization in acinar cells.

Atrial natriuretic factor (ANF) binding sites have been recently demonstrated to be present in exocrine pancreas by an in vitro autoradiographic approach. An autoradiographic study was carried out to identify the exocrine cells containing ANF binding sites and to monitor the fate of 125I-labeled ANF in acinar cells after removal of pancreas at specific time intervals (1-30 min) after intravenous administration. At the light microscopic level, silver grains were found over acinar and centroacinar cells. Concomitant injection of an excess of unlabeled ANF inhibited the binding of labeled peptide by approximately 60%. At the electron microscopic level, the time-course study in acinar cells has revealed that of the cell compartments examined, plasma membrane, Golgi apparatus, mitochondria, and zymogen granules, the nucleus had distinct labeling patterns. Plasma membrane was maximally labeled 1 and 2 min after injection with 125I-ANF. Golgi apparatus was significantly labeled from 2 to 30 min after injection, mitochondria from 1 to 30 min after injection, zymogen granules at 1 and 15 min, and the nucleus only at 30 min. The lysosomal compartment was not labeled during the 30-min observation period. These results suggest that after binding to the plasma membrane, ANF is rapidly internalized and distributed to the intracellular organelles as a function of time. Labeling of the zymogen granules suggests that they may bind ANF and that the atrial peptide may be secreted by acinar cells. The significance of association of radioactivity with mitochondria and nuclei remains to be elucidated but may represent intracellular sites of action of ANF complementary to those on plasma membranes.

Animals↗
PubMed1988

Evidence for a direct action of neuropeptide Y in the rat pituitary gland.

Neuropeptide Y (NPY) has recently been localized in the rat hypothalamus. In order to evaluate the putative effects of NPY on pituitary function, its action was examined on anterior pituitary cells in culture. Also, an immunocytochemical method was used with the aim of localizing endogenous NPY-like material at the cellular and subcellular levels of the pituitary gland. In vitro studies using dispersed anterior pituitary cells indicated that NPY (10(-6) to 10(-9)M) increased the secretion of luteinizing hormone, growth hormone and prolactin, whereas beta-lipotropin hormone and thyrotropin secretions were not affected. The presence of endogenous NPY was demonstrated in gonadotrophs, somatotrophs, corticotrophs and some lactotrophs, but not in thyrotrophs. In immunoreactive cells, NPY-like material was detected in the cytoplasmic matrix, in the secretory granules and in the nucleus distributed primarily in the euchromatin, in the vicinity of the heterochromatin. NPY-like immunoreactivity was also observed at the plasma membrane but only scarcely. These biochemical and immunocytochemical results indicate that NPY may play a direct regulatory role in adenohypophyseal secretion.

Animals↗
PubMed1988

Synthesis, internalization, and localization of atrial natriuretic peptide in rat adrenal medulla.

Some, though not all studies, have indicated that atrial natriuretic peptide (ANP) can bind to adrenal medullary cells. ANP-like immunoreactivity (ANP-LI) has also been identified in catecholamine-secreting cells. Together, these findings suggest that ANP may be taken up and/or synthesized in the adrenal medulla. The present study was designed to ascertain, by in situ hybridization, whether adrenal chromaffin cells could synthesize ANP, to define by an in vivo ultrastructural autoradiographic approach, whether ANP could, in fact, bind to rat adrenal medulla cells, to determine whether there was a cellular [noradrenaline (NA) vs. adrenaline (A)] selectivity in the binding process, and to establish whether extracellular [125I]ANP could be internalized by these cells. The cellular and subcellular distribution of endogenous ANP-LI was also investigated in both cell types by cryoultramicrotomy and immunocytochemical approaches. The in situ hybridization studies indicate the presence of mRNA to ANP in about 15% of adrenal medullary cells. Intravenous injection of [125I]ANP resulted in a 3-fold, preferential and specific radiolabeling of A-as compared to NA-containing cells. In A-containing cells, plasma membranes were significantly labeled 2 and 5 min post injection; cytoplasmic matrix, mitochondria, and secretory granules throughout the time course studied (1-30 min post injection). Lysosomes, rough endoplasmic reticulum, Golgi apparatus, and nuclei were not labeled. ANP-LI was identified in both NA- and A-containing cells; in the former, it was almost exclusively localized in secretory vesicles, in the latter it was detected in plasma membranes, cytoplasmic matrix, nuclear euchromatin, some mitochondria and relatively fewer granules than in NA-containing cells. The findings suggest that ANP may be synthesized primarily in NA-containing cells and that A-containing cells primarily bind and internalize the extracellular (endogenous or exogenous) atrial peptide. The data suggest that ANP secreted by adrenal medullary chromaffin cells may have distal paracrine actions or interactions with coreleased catecholamines and neuropeptides. Binding and internalization may reflect an action of ANP on the secretory function of A-containing cells.

Adrenal Medulla↗
PubMed1988

Ultrastructural evidence for oxytocin in the rat anterior pituitary gland.

OT is synthetized in the hypothalamus. These neurons project to the posterior lobe of the pituitary and the external zone of the median eminence. In order to localize OT in the male rat anterior pituitary we have used immunocytochemistry on ultra-thin sections in target cell(s) obtained by cryoultramicrotomy. OT-like immunoreactivity was observed in lactotropes only. No immunoreactivity was observed in gonadotropes, somatotropes, corticotropes or thyrotropes. In lactotropes, immunoreactivity was localized at the plasma membrane level, in the cytoplasmic matrix and around the secretory granules, but not in the other organelles, and in the nucleus. No reaction was observed by using either non-immune serum or anti-OT serum incubated with OT. No modification of OT-like immunoreactivity was observed by using anti-OT serum incubated with heterologous peptides. These results 1) provide immunocytological evidence for the presence of OT in the anterior pituitary gland; 2) indicate the presence of this peptide in one particular cell type, and 3) support the hypothesis that OT could have a direct participation in the regulation of the PRL release.

Animals↗
PubMed1987

In situ hybridization to rat brain and pituitary gland of growth hormone cDNA.

Using in situ hybridization, we have investigated the presence of mRNA coding for growth hormone (GH) in the rat brain. Using 32P-labeled GH cDNA as a probe, the pituitary gland showed hybridization in unfixed sections. Using 3H-labeled GH cDNA hybridized to fixed sections, only cells in the anterior pituitary were labeled in good agreement with the localization of somatotropes. In the brain, wide zones were labeled with 32P-GH cDNA: the caudate putamen, the striatum, the ventral thalamus, the formatio reticularis and the basal cortex. With the 3H GH-cDNA probe, more discrete regions of the brain showed hybridization: the basal cortex, the outside part of the hippocampus and the caudate putamen.

Animals↗
PubMed1987

Binding and internalization of native gonadoliberin (GnRH) by anterior pituitary gonadotrophs of the rat. A quantitative autoradiographic study after cryoultramicrotomy.

To identify anterior pituitary cell types containing GnRH binding sites and to study the internalization process of this peptide by target cells under physiological conditions, autoradiography was performed on rat anterior pituitaries removed at specific time intervals (2-60 min) after intravenous injection of mono-radioiodinated 125I-GnRH into intact males. At electron-microscopic level, gonadotrophs and lactotrophs appeared to contain silver grains. Concomitant administration of an excess of unlabeled GnRH with the radioiodinated hormone prevented this localization indicating the specificity of the reaction. The time-course study in gonadotrophs showed that 2 min after injection silver grains could be found over the plasma membrane, secretory granules and nuclear membrane. Similar results were observed 5 and 15 min after injection. Extensive label was observed over the nucleus and nuclear membrane 15 to 60 min after injection. The injection of a radioiodinated GnRH agonist [D-Trp6, Pro9 (Net), DesGly10]-GnRH produced comparable results. In contrast, the injection of 125I-[D-pGlu1, D-Phe2, Trp3,6]-GnRH, an antagonist of GnRH, produced positive labeling only at the plasma membrane without internalization. These results indicate that, after binding with receptors on the plasma membrane, GnRH is rapidly internalized, accumulating in secretory granules, and localizing over the nuclear membrane and later, in the nucleus. Association of radioactivity with secretory granules could be related to a specific action of GnRH at this level or to receptor recycling, and presence of label in the nucleus may be related to stimulation of neosynthesis of LH and GnRH receptors.

Animals↗
PubMed1987

A new technique for mapping epitope specificities of monoclonal antibodies using quasi-elastic light scattering spectroscopy.

Quasi-elastic light scattering (QLS) was used to determine relative epitope specificities of a group of anti-bovine serum albumin (BSA) monoclonal antibodies (MAb). QLS is a non-invasive technique which can determine the mean size and size distributions of macromolecular scatterers by analysis of the fluctuations in the intensity of laser light scattering. When two MAbs are mixed together with antigen, QLS detects the complex formation which results from the Ag-Ab reaction, and can easily distinguish between the large complexes formed by interaction of non-competitive MAbs and the smaller complexes formed by competitive MAbs. In this report, the competitive or non-competitive behavior of six anti-BSA MAbs were assessed by radioimmunoassay (RIA) and QLS analysis. The results obtained by QLS analysis confirmed the RIA findings indicating that the six MAbs examined can be categorized into three distinct, non-interacting groups. QLS analysis represents a simple, and extremely rapid technique for epitope mapping studies.

Antibodies, Monoclonal↗
PubMed1987

Atrial natriuretic factor and exocrine pancreas: effects on the secretory process.

Immunoreactive atrial natriuretic factor (ANF) has recently been identified in pancreatic acinar cells. The current study therefore, was, designed to ascertain whether the atrial peptide exercises any biological effects on the exocrine pancreas. When isolated rat pancreatic acinar cells were incubated with rat ANF (8-33), a concentration-dependent increase in cGMP synthesis was observed (EC50 about 5 X 10(-9) M) with the peak response occurring within 2.5 min of exposure of the cells to the peptide. ANF did not affect basal or secretagogue (carbachol +/- DbcAMP, CCK-OP, or forskolin)-induced amylase secretion from acinar cells, nor did it affect [3H]thymidine incorporation into DNA or [3H]leucine incorporation into trichloroacetic acid-precipitable protein. ANF was also infused intravenously (0.01-0.25 micrograms/min) in rabbits with cannulated pancreatic ducts, and the peptide stimulated a dose-dependent secretion of cGMP into pancreatic juice. ANF, by itself, did not affect protein or fluid secretion from rabbit pancreas; when co-infused with secretin (0.1 CU/min), which is not an acinar cell secretagogue in rabbits, ANF increased fluid secretion in one of three animals tested. The data suggest that acinar cells possess functional ANF receptors, whose activation results in both synthesis and secretion of cGMP. Intra-acinar cell cGMP is not involved in the enzyme secretory process. Secreted nucleotide is not co-released with digestive hydrolases, and does not, by itself, appear to have an intraluminal effect on fluid secretion. The mechanism by which acinar cells secrete cGMP into pancreatic juice as well as the biological significance of intracellular (acinar), and extracellular (intraluminal) cGMP remain to be elucidated.

Amylases↗