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G Pelletier

Publications and source records attributed to G Pelletier.

At least 217 records · Page 12Linked to original sources
PubMed1994

Ogura cytoplasmic male-sterility (CMS)-associated orf138 is translated into a mitochondrial membrane polypeptide in male-sterile Brassica cybrids.

Transcription of a putative mitochondrial gene (orf138) has previously been correlated with Ogura cytoplasmic male-sterility (CMS) in rapeseed cybrids. In this paper, studies performed on a Brassica cybrid with a different organization of the orf138 locus confirm this association. We also show that mitochondria isolated from male-sterile rapeseed plants synthesize a polypeptide of 19 kDa, which is absent in fertile revertants. Antibodies against a glutathione S-transferase-ORF138 fusion protein were raised to establish that this 19 kDa polypeptide is the product of orf138. The anti-ORF138 serum was used to demonstrate that the orf138 translation product occurs only in sterile cybrids and co-purifies with the mitochondrial membrane fraction.

Blotting, Western↗
PubMed1994

Three-dimensional type I collagen gel system for the study of osteoblastic metastases produced by metastatic prostate cancer.

We developed a three-dimensional type I collagen gel cell culture system that allows coculturing of human MG-63 osteoblast-like cells and various human cancer cells. Inoculation of human PC-3 metastatic prostate cancer cells into this type I collagen gel containing human MG-63 osteoblast-like cells produced an osteoblastic-like reaction that presented as an increased number of MG-63 cells and increased density of type I collagen around MG-63 cells adjacent to inoculated PC-3 cells by microscope analysis. Under identical experimental conditions, inoculation of cell-free medium, human KLE endometrial adenocarcinoma cells, and Calu-1 lung cancer cells did not produce this blastic-like reaction. In situ hybridization documented the uniform expression of insulin-like growth factor I (IGF-I) and of urokinase-type plasminogen activator (uPA) mRNA in MG-63 and PC-3 cells separately cultured in this substrata. The uniform expression of uPA was also documented by immunocytochemistry using a monoclonal and a polyclonal antihuman uPA antibody. The relative expression of uPA was higher in PC-3 cells than in MG-63, KLE, and Calu-1 cancer cells. We conclude that this novel cell culture system may become a useful model to study the pathophysiology of the osteoblastic reaction in vitro.

Adenocarcinoma↗
PubMed1994

Localization of 3 beta-hydroxysteroid dehydrogenase in rat brain as studied by in situ hybridization.

Recent evidence suggests that brain cells can synthesize steroids de novo. The steroidogenic enzyme 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) gene family of enzymes is involved in the biosynthesis of all classes of active steroids. In the present study, we have used in situ hybridization performed with a 35S-labeled cDNA-encoding rat type I 3 beta-HSD to localize the expression of member(s) of this gene family in the adult rat brain. A specific hybridization signal could only be detected in a restricted area of the medulla ventrally and laterally bordering the fourth ventricle. This area contains the nucleus propositus hypoglossi as well as some vestibular nuclei: the nucleus vestibularis medialis, the nucleus vestibularis lateralis, and the nucleus vestibularis spinalis. At high resolution, the silver grains representing a hybridization signal were detected exclusively in neurons. The present data strongly suggest that a restricted population of neurons might be involved in steroid biosynthesis in the adult rat brain. The role of the member(s) of the rat 3 beta-HSD family in the nuclei of two cranial nerves remains to be investigated.

3-Hydroxysteroid Dehydrogenases↗
PubMed1994

Immunocytochemical localization of 5 alpha-reductase in rat brain.

The presence of steroid 5 alpha-reductase in brain tissue is well-documented. In order to determine with precision the brain areas and nerve cell types containing this enzyme in adult rat, we have studied its immunocytochemical localization using antibodies to the sequence peptide comprising amino acids 227 to 240 of the enzyme. At the light microscopic level, 5 alpha-reductase immunoreactive material appeared to be present only in glial and ependymal cells, including tanycytes. Immunolabeled cells were found in high concentration in the hypothalamus, thalamus, hippocampus, cerebral cortex, cerebral cortex, and circumventricular organs. Immunoelectron microscopic studies demonstrated that immunoreactive material was diffusely distributed throughout the cytoplasm of glial and ependymal cells, thus confirming the association of 5 alpha-reductase immunoreactivity with nonneuronal cells. These data suggest that glial cells play an important role in the conversion of testosterone to dihydrotestosterone or progesterone to dihydroprogesterone under in vivo conditions in adult rat.

Animals↗
PubMed1994

Characterization of some major identity elements in plant alanine and phenylalanine transfer RNAs.

Alanine and phenylalanine tRNA sequences were amplified by PCR from Arabidopsis thaliana nuclear DNA using degenerate oligonucleotides which introduced specific mutations into the acceptor stem. The aminoacylation of T7 RNA polymerase transcripts of these sequences was investigated in vitro using partially purified bean alanyl- or phenylalanyl-tRNA synthetase. In parallel, the in vivo activity of amber suppressor derivatives of these tRNAs was investigated in transient expression assays in tobacco protoplasts using a beta-glucuronidase (GUS) reporter gene containing a premature amber stop codon. The results show that mutation of the G3:U70 base pair to G3:C70 blocks aminoacylation of plant alanine tRNA, whilst conversion of the G3:C70 pair normally found in plant tRNA(Phe) to G3:U70 enables the mutated tRNA(Phe) to be a good substrate for alanyl-tRNA synthetase and impairs its aminoacylation with phenylalanine. In addition, the amber suppressor derivative of wild-type tRNA(Phe) showed very little suppressor activity in vivo, and was poorly aminoacylated with phenylalanine in vitro, suggesting that the anticodon is a major identity determinant for tRNA(Phe) in plant cells.

Alanine-tRNA Ligase↗
PubMed1994

Localization of diazepam-binding inhibitor and peripheral type benzodiazepine binding sites in the rat ovary.

Diazepam-binding inhibitor (DBI) is the precursor of a family of peptides, including an octadecaneuropeptide (ODN), that share with DBI the ability to specifically displace benzodiazepines (BZD) from their receptors. An association of ODN with the peripheral type BZD receptors (PBR) has been reported in the brain and a few peripheral tissues. In order to investigate whether DBI and PBR are present in ovarian tissue, we have localized DBI by means of immunocytochemistry, in situ hybridization and autoradiography of PBR in the rat ovary. Immunocytochemical localization was achieved by means of rabbit antibodies developed against rat ODN. Immunostaining was located in the cytoplasm of the theca interna, corpus luteum and interstitial gland cells, but not in the granulosa cells. Hybridization signal obtained following in situ hybridization with a [35S]-labelled single-stranded RNA probe complementary to DBI mRNA was observed in all the steroid-secreting cells, including granulosa cells of developing and mature follicles. Autoradiographic localization of PBR obtained by incubating ovary sections with [3H] PK11195, a ligand selective for PBR, revealed the presence of specific labelling in all the steroid-secreting cells. These results, which demonstrate for the first time that the ovarian steroid-secreting cells contain both PBR and its endogenous ligand, suggest that the BZD receptor might be involved in the regulation of ovarian function.

Animals↗
PubMed1994

Occurrence of 5c,8c,11c,15t-eicosatetraenoic acid and other unusual polyunsaturated fatty acids in rats fed partially hydrogenated canola oil.

Uncommon cis and trans fatty acids can be desaturated and elongated to produce unusual C18 and C20 polyunsaturated fatty acids in animal tissues. In the present study we examined the formation of such metabolites derived from cis and trans isomers of oleic and linoleic acids of partially hydrogenated vegetable oil origin in rats. For two months, adult male rats were fed a partially hydrogenated canola oil diet containing moderately high levels of trans fatty acids (9.6 energy%) and an adequate level of linoleic acid (1.46 energy%). Analysis of the phospholipid (PL) fatty acids of liver, heart, serum and brain showed no new C18 polyunsaturated fatty acids, except for those uncommon 18:2 isomers originating from the diet. However, minor levels (each < 0.3% PL fatty acids) of six unusual C20 polyunsaturated fatty acids were detected in the tissues examined, except in brain PL. Identification of their structures indicated that the dietary 9c,13t-18:2 isomer, which is the major trans polyunsaturated fatty acid in partially hydrogenated vegetable oils, was desaturated and elongated to 5c,8c,11c,15t-20:4, possibly by the same pathway that is operative for linoleic acid. Furthermore, dietary 12c-18:1 was converted to 8c,14c-20:2 and 5c,8c,14c-20:3; dietary 9c,12t-18:2 metabolized to 11c,14t-20:2 and 5c,8c,11c14t-20:4, and dietary 9t,12c to 11t,14c-20:2. These results suggested that of all the possible isomers of oleic and linoleic acids in partially hydrogenated vegetable oils, 12c-18:1, 9c,13t-18:2, 9c,12t-18:2 and 9t,12c-18:2 are the preferred substrates for desaturation and elongation in rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Role of neuropeptide Y in the regulation of gonadotropin-releasing hormone gene expression in the rat preoptic area.

It is well documented that neuropeptide Y (NPY) is involved in the regulation of the hypothalamo-pituitary gonadal axis. In order to determine the influence of NPY on the biosynthesis of GnRH, we have studied the effects of NPY and some NPY analogs on GnRH gene expression in neurons in the male rat preoptic area (POA). The following peptides NPY, peptide YY (PYY), [Leu31,Pro34]NPY (a Y1 receptor agonist) and NPY13-36 (a Y2 receptor agonist) were injected into the left lateral ventricle of adult castrated male rats. In another series of experiments, the same peptides were administered intravenously (IV). All the animals were perfused with 4% paraformaldehyde 4 hours after injection. Cryostat sections through the POA were processed for quantitative in situ hybridization. The intracerebroventricular injection of PYY, NPY and [Leu31,Pro34]NPY induced a marked increase in the number of grains overlying the labelled neurons (20 to 45% over control). On the other hand, the Y2 receptor agonist NPY13-36 did not influence mRNA levels. Similar results were obtained following the i.v. administration although the magnitude of the stimulating effect was less important (12 to 20% over control). These data then strongly suggest that NPY positively regulates the genetic expression of GnRH in neurons via the Y1 NPY receptor subtype.

Animals↗
PubMed1994

Negative regulation of proopiomelanocortin gene expression by GABAA receptor activation in the rat arcuate nucleus.

Activation of the GABAA-benzodiazepine receptor complex has previously been shown to inhibit the release proopiomelanocortin (POMC)-related peptides from the hypothalamus and to decrease mRNA levels in POMC neurons in the arcuate nucleus. To learn more about the precise role of the GABAergic system in POMC neuron regulation, we studied the effects of the administration (2 days) of the GABAA receptor agonist muscimol and the central-type barbiturate receptor agonist pentobarbital on POMC mRNA levels measured by in situ hybridization. Treatment with pentobarbital produced a 12% decrease in the hybridization signal. Similarly, muscimol treatment decreased the signal by 20%. The concomitant administration of the two GABAA receptor agonists resulted in a decrease (28%) of mRNA levels that was significantly more marked than that induced by pentobarbital or muscimol alone. The present results, together with previous data from our laboratory, indicate that different activators of the GABAA receptor complex, including barbiturates, can negatively regulate POMC neuronal activity in the rat arcuate nucleus.

Animals↗
PubMed1994

Role of serotonin in the regulation of prolactin gene expression in the male rat as evaluated by in situ hybridization.

It is well documented that serotonin (5-HT) is involved in the regulation of prolactin (PRL) release as a stimulator. To evaluate the role of 5-HT on PRL gene expression, we have investigated the effects of repeated administration (during 2 days) of 5-HT, the 5-HT1 + 2 receptor antagonist methysergide, the 5-HT2 receptor antagonist ketanserin, and a combination of 5-HT and methysergide on PRL mRNA levels as measured by in situ hybridization in the adult male rat. The treatment with 5-HT induced a 14% increase in the hybridization signal, although the administration of methysergide produced a small reduction (5%) in PRL mRNA levels. On the other hand, ketanserin had no effect on the hybridization signal. The stimulatory effect of 5-HT was completely prevented by the concomitant administration of methysergide. The present results, together with previous data, indicate that 5-HT exerts a positive tonic influence not only on the release of PRL but on the biosynthesis of the hormone as evaluated by mRNA level measurements. They also strongly suggest that the role of 5-HT in the regulation of PRL secretion is mediated via activation of 5-HT1 receptors, although a dopaminergic effect of methysergide on PRL secretion cannot be totally excluded.

Animals↗
PubMed1994

Structure, regulation and role of 3 beta-hydroxysteroid dehydrogenase, 17 beta-hydroxysteroid dehydrogenase and aromatase enzymes in the formation of sex steroids in classical and peripheral intracrine tissues.

In addition to the classical steroidogenic tissues, namely the ovaries, testes, adrenals and placenta, a large series of human peripheral tissues possess all the enzymatic systems required for the formation of active androgens and oestrogens from a relatively large supply of precursor steroids provided by the adrenals. This chapter describes the structure, function, tissue-specific expression and regulation of the 3 beta-HSD and 17 beta-HSD gene families as well as some information about the aromatase gene. While, so far, most therapeutic approaches have been aimed and limited at controlling steroid formation by the classical steroidogenic tissues, it is clear that major efforts should now be turned towards intracrinology in order to understand better the physiological mechanisms controlling local steroid formation in peripheral target tissues and thus be in a position to develop novel therapeutic approaches that take into account the high proportion of steroids that are made locally and are responsible for the growth and function of normal as well as cancerous tissue.

17-Hydroxysteroid Dehydrogenases↗
PubMed1994

Management of clinically relevant bleeding following endoscopic sphincterotomy.

We report here on our experience of clinically relevant bleeding after endoscopic sphincterotomy (ES). Relevant bleeding was defined by the occurrence of (a) hematemesis or melena and (b) at least a two-point drop in hemoglobin, with no other bleeding source on endoscopy. These two criteria were met in 16 patients between 1983 and 1992. They represented 0.65% of all ES procedures performed during this period. Bleeding occurred immediately after ES in five cases, and was delayed in 11 cases from one to eight days (mean two days). Patients were retrospectively classified into three groups according to the severity of bleeding and subsequent clinical management. In six cases (group 1), bleeding developed slowly without shock and stopped spontaneously. In five cases (group 2), bleeding developed rapidly with melena and a drop in hemoglobin, but without shock. These patients were successfully managed with sclerotherapy without any further complications. The five patients in Group 3 had brisk bleeding with hematemesis and shock. Endoscopic hemostasis could not be performed; emergency arteriography disclosed active bleeding in four patients, and embolization of the gastroduodenal artery was performed. Bleeding stopped in all patients. Billroth II anastomosis appeared to be the only factor associated with an increased risk of clinically relevant bleeding. It was possible to control bleeding following ES using endoscopic or angiographic hemostasis, surgery being avoided in all cases.

Adult↗
PubMed1994

Transport of mannose by an inducible phosphoenolpyruvate:fructose phosphotransferase system in Streptococcus salivarius.

Streptococcus salivarius transports mannose by a phosphoenolpyruvate:sugar phosphotransferase system (PTS) which consists of a membrane Enzyme II and two forms of Enzyme III (IIIMan) with molecular masses of 38.9 kDa (IIIManH) and 35.2 kDa (IIIManL) respectively. Using a pseudorevertant (strain 57P) isolated from a IIIManL-deficient spontaneous mutant unable to grow on mannose, we demonstrated that S. salivarius could also transport mannose by an inducible fructose PTS. This PTS phosphorylated fructose at the C-1 position with a high affinity (10 microM) and mannose at the C-6 position with a low affinity (200 microM). Derepression of this system in some IIIManL-deficient mutants would explain their ability to grow on mannose.

Binding, Competitive↗
PubMed1994

Pituitary adenylate cyclase-activating polypeptide stimulates both adrenocortical cells and chromaffin cells in the frog adrenal gland.

In a previous report, we have shown that frog pituitary adenylate cyclase-activating polypeptide (fPACAP38) is a potent stimulator of corticosteroid secretion by frog adrenal slices in vitro. The aim of the present study was to determine the mode of action of PACAP on the frog adrenal gland. Immunoelectron microscopic labeling revealed that PACAP-like immunoreactivity is present in electron-dense vesicles within nerve endings located in the vicinity of both adrenocortical and chromaffin cells. Exposure of dispersed adrenal cells to fPACAP38 caused stimulation of corticosteroid secretion. Labeling of cultured adrenal cells with [125I]PACAP27 revealed the existence of PACAP-binding sites on both adrenocortical and chromaffin cells. Saturation and competition experiments showed the occurrence of high affinity and selective receptors for fPACAP38 on cultured adrenal cells. fPACAP38 (10(-8)-10(-5) M) provoked a dose-dependent stimulation of cAMP production by frog adrenal slices. Microflurimetric studies demonstrated that fPACAP38 induced a substantial elevation of the intracellular calcium concentration in both adrenocortical and chromaffin cells. The present results indicate that in the frog adrenal gland, PACAP fibers innervate both adrenocortical and chromaffin cells. The data show the presence of PACAP receptors on the two cell types. PACAP exerts a direct stimulatory effect on corticosteroid-producing cells. This effect is probably mediated through stimulation of adenylyl cyclase activity and/or augmentation of intracellular Ca2+. PACAP also increases intracellular Ca2+ in chromaffin cells. These data suggest that PACAP, released locally in the adrenal gland, acts as a neuroendocrine factor, regulating the activity of adrenocortical and chromaffin cells.

Adrenal Cortex↗
PubMed1994

Long-term effects of human growth hormone-releasing hormone and photoperiod on hormone release and puberty in dairy heifers.

Forty-eight Holstein dairy heifers (98.9 kg BW; 3 mo old) were subjected for 246 d to twice-daily s.c. injections of saline (CTL) or human growth hormone-releasing hormone (GRH; 5 micrograms/kg BW) and to photoperiods of 8 h of light (L): 16 h of dark (D) or 16L:8D according to a 2 x 2 factorial arrangement of treatments. Jugular blood samples were collected from 16 heifers at 3, 4, 8, and 11 mo of age to monitor prolactin, growth hormone, and estradiol-17 beta. Plasma progesterone concentrations were monitored weekly in all heifers as an index of puberty (> 1 ng/mL). Growth hormone release was induced by GRH (P < .001) throughout the trial; area under the GH curve (AUC) averaged 1,582 vs 3,643 ng.min-1.mL-1 in CTL vs GRH heifers. However, GRH-induced GH response was less (P < .05) after the second daily injection. There was also an interaction (P = .08) between GRH, photoperiod, and days of treatment on GRH-induced GH response; AUC was greater in GRH-16L:8D than in GRH-8L:16D heifers at 3 mo but less at 8 mo of age. The PRL concentrations were similar for both photoperiods at 3 mo (36.4 vs 41.7 ng/mL) and 8 mo (16.2 vs 12.8 ng/mL) of age but were greater in 16L:8D vs 8L:16D heifers at 4 mo (18.4 vs 39.3 ng/mL) and 11 mo (26.3 vs 44.1 ng/mL) of age (photoperiod x day interaction, P < .001). Photoperiod of 16L:8D vs 8L:16D reduced (P < .01) weight at puberty in CTL heifers (251 vs 303 kg BW) and to a lesser extent in GRH-treated heifers (271 vs 284 kg BW; GRH x photoperiod interaction, P = .10). In conclusion, GH response is maintained throughout 8 mo of GRH treatment, and a 16L:8D photoperiod will reduce age and weight at puberty in heifers. Furthermore, refractoriness to photoperiod-induced PRL changes was detected.

Aging↗