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F Demay

Publications and source records attributed to F Demay.

9 recordsLinked to original sources
PubMed2001

Steroid-independent activation of ER by GnRH in gonadotrope pituitary cells.

In the rat pituitary gland the mechanism responsible for ERalpha regulation has not been fully elucidated. Using transient transfection assays in alphaT3-1 cells, a cell line of gonadotrope origin, we show that GnRH stimulates estrogen response element-containing promoters in an estrogen-independent manner. This effect was strictly ER and GnRH receptor dependent, as no activation of the reporter gene was observed in presence of the anti-estrogen ICI 182,780 or a GnRH antagonist. These data suggest that the GnRH-triggered signaling pathway results in 17beta-estradiol-independent trans-activation of the ERalpha in alphaT3-1 cells. Furthermore, an additive activation was achieved when cells were treated with both GnRH and 17beta-estradiol. In primary pituitary cells, GnRH alone (100 nM) did not cause a significant stimulation of reporter gene activity, presumingly due to the low amount of gonadotropes. Interestingly, the combination of 17beta-estradiol and GnRH resulted in a significant increase in ERalpha trans-activation compared with that in cells treated with 17beta-estradiol alone. This enhancement was prevented by ICI 182,780, showing an ERalpha requirement. Moreover, we show that the effects of GnRH on ERalpha transcriptional activity in gonadotrope cell lines are mediated by the PKC/MAPK pathway. In conclusion, our data demonstrate that GnRH is an important signal in the regulation of ERalpha trans-activation in gonadotrope cells.

Animals↗
PubMed1998

In vitro induction of specific cytotoxic T lymphocytes using recombinant single-chain MHC class I/peptide complexes.

We have previously described the production and purification of a murine single-chain, soluble recombinant major histocompatibility complex (MHC) class I molecule (SC-Kd). A similar strategy was devised to produce a recombinant HLA-A2.1 (SC-A2) molecule. The latter was composed of the first three domains of the HLA-A2.1 heavy chain connected to human beta 2-microglobulin through a spacer of 15 amino acids. Immunoaffinity-purified SC-A2 molecules-were correctly folded and biologically functional. They specifically bound HLA-A2-restricted peptides and induced a peptide-specific cytotoxic T lymphocyte (CTL) clone to proliferate and secrete interleukin-2. The ability of murine and human SC-MHC molecules to elicit primary CTLs in vitro was next investigated. When coated in high density onto beads, complexes of antigenic peptide and SC-Kd or SC-A2 molecules efficiently induced a specific primary CTL response in vitro. Furthermore, the structural features of these CTLs were characterized by T cell receptor-beta chain analysis, which revealed rearrangements very similar, if not identical, to those found in CTLs generated by in vivo immunization. Such single-chain, soluble recombinant MHC class I molecules should provide a useful tool in particular for peptide binding assays and for in vitro primary CTL induction to identify immunogenic peptides such as those derived from known tumor-associated antigens.

Animals↗
PubMed1996

Cell-specific mechanisms of estrogen receptor in the pituitary gland.

Analysis of the mechanism of action of estrogen receptor shows protein and mRNA polymorphism within distinct pituitary receptor-positive cells. The lactotropes exhibit unique properties in these mechanisms that distinguish them from gonadotropes. Therefore, this cell type constitutes an especially interesting model in the male as well as in the female for estrogen receptor studies.

Animals↗
PubMed1996

Effect of gonadotropin-releasing hormone on estrogen receptor messenger ribonucleic acid level in perifused pituitary cells.

1. We examined the potential effect of GnRH pulses on pituitary estrogen receptor mRNA level. 2. The treatment of perifused pituitary cell aggregates with four hourly pulses of GnRH (10nM/1 min/h) resulted in a marked increase in the steady-state level of ER mRNA (25% vs unstimulated control, n = 3). 3. No changes were observed for the LH beta mRNA. Data suggest, for the first time, that a cross-talk between the GnRH and nuclear ER may occur in the gonadotrope cells.

Animals↗
PubMed1996

Sex- and cell-specific expression of an estrogen receptor isoform in the pituitary gland.

The presence of multiple monomeric forms has been described for the estrogen receptor (ER) in the pituitary gland. We analyzed ER mRNA forms in male and female rat pituitary. A single 6.2-kb ER mRNA species was detected in the male rat pituitary, whereas the female rat pituitary exhibited two ER mRNA forms of 6.2 and 5.5 kb, respectively. The 6.2-kb mRNA was present throughout the different stages of the estrous cycle, while the 5.5-kb mRNA appeared to be restricted to proestrus, suggesting an acute regulation of ER transcription at this stage. The 5.5-kb ER mRNA could be rapidly induced either by 17 beta-estradiol replacement in ovariectomized adult female rats or by priming immature rats with pregnant-mare serum gonadotropin. Using enriched cell populations, an inverse and strong correlation was established between the presence of the 5.5-kb ER mRNA form and the number of gonadotropes. Conversely, the localization of the 5.5-kb mRNA form was demonstrated in lactotrope populations. In order to elucidate the structural modifications in the transiently expressed ER mRNA, a series of reverse-transcriptase polymerase chain reaction amplifications was carried out using several pairs of primers corresponding to the entire ER-coding region. The data showed that no alternative splicing was occurring in the ER-coding region involving a potential role of either 3'- or 5'-untranslated regions. Thus, ER presents a 17 beta-estradiol-dependent transcriptional mechanism triggered on proestrous day and specific to the female lactotropes.

Animals↗
PubMed1980

The effect of piracetam on volunteers in a low-pressure tank.

Twelve volunteers were subjected to a double-blind crossover test to evaluate their concentration in conditions of hypoxemia after treatment with piracetam or with a placebo. Piracetam was administered in doses of 4.8 g per day for 4 days and 7.2 g on the 5th day. The speed with which the test was carried out was not influenced to a statistically significant degree but the proportion of errors was. This effect of piracetam was more pronounced for the longer-lasting periods of hyposemia.

Adult↗
PubMedDate not supplied

Quantitative PCR with colorimetric detection.

We describe a quantitative polymerase chain reaction (PCR) technique with a colorimetric read-out, which enables quantitation of various sequences on a single microplate within one day. Using the quantitation of human IL10 and beta-actin transcripts, we show that this technique provides better reproducibility, as results are derived from a series of PCRs made with various amounts of standard. This also enables the control of the consistency of the data and elimination of some artifactual results.

Actins↗