Biomedical subjects

Florian Guillou

Publications and source records attributed to Florian Guillou.

23 records · Page 2Linked to original sources

PubMed2002

Sertoli and granulosa cell-specific Cre recombinase activity in transgenic mice.

We have established transgenic mice expressing the Cre recombinase under the control of the anti-Müllerian hormone (AMH) gene promoter. Cre activity and specificity were evaluated by different means. In AMH-Cre mice, expression of the Cre recombinase mRNA was confined to the testis and ovary. AMH-Cre mice were crossed with reporter transgenic lines and the offspring exhibited Cre-mediated recombination only in the testis and the ovary. In male, histochemical analysis indicated that recombination occurred in every Sertoli cells. In female, Cre-mediated recombination was restricted to granulosa cells, but the protein was not evenly active in every cells. From these results, we conclude that potentially, this transgenic line possessing AMH promoter-driven expression of the Cre recombinase is a powerful tool to delete genes in Sertoli cells only, in order to study Sertoli cell gene function during mammalian spermatogenesis.

Animals↗

PubMed2002

Alternative splicing in the brain of mice and rats generates transferrin transcripts lacking, as in humans, the signal peptide sequence.

Transferrin (Tf), the iron-transport protein of vertebrate serum, is mainly synthesized in hepatocytes but is also found in other cell-types including oligodendrocytes. Our laboratory has characterized in a human oligodendrial cell line the presence of a new Tf transcript containing an alternative exon 1b replacing the classical exon 1 and conducting to the elimination of the signal peptide sequence. In this manuscript, we show by RT-PCR and 5'-RACE experiments that alternative transcripts also exist in mouse and rat and are found in brain mRNA preparations. Mouse alternative first exon is homologous to human exon 1b while rat Tf gene was found to use a new first exon named 1c. In all species, the alternative transcript does not contain the signal peptide sequence and possibly encode for a Tf protein devoid of signal peptide showing that this phenomenon is not restricted to human gene. We also present genomic sequence data from the previously unknown 5' genomic rat region, which allowed the alignment of the alternative exons 1 in the three species.

Alternative Splicing↗

PubMed2002

G protein-coupled receptor kinases and beta arrestins are relocalized and attenuate cyclic 3',5'-adenosine monophosphate response to follicle-stimulating hormone in rat primary Sertoli cells.

The FSH receptor (FSH-R) is a member of the rhodopsin-like subfamily of G protein-coupled receptors that undergoes homologous desensitization upon agonist stimulation. In immortalized cell lines overexpressing the FSH-R, G protein-coupled receptor kinases (GRKs) and beta-arrestins are involved in the phosphorylation, uncoupling, and internalization of this receptor. In an effort to appreciate the physiological relevance of GRK/beta-arrestin actions in natural FSH-R-bearing cells, we used primary rat Sertoli cells as a model. GRK2, -3, -5, -6a, and -6b and beta-arrestins 1 and 2 were expressed in primary rat Sertoli cells. Overexpression of these different GRKs and beta-arrestins in primary rat Sertoli cells significantly attenuated the FSH-induced cAMP response, and FSH rapidly triggered a relocalization of endogenously expressed GRK2, -3, -5, and -6 and beta-arrestins 1 and 2 from the cytosol to the membranes. These results highlight the relationship existing between the GRK/beta-arrestin regulatory system and the FSH-R signaling machinery in a physiological model.

Animals↗

PubMed2002

Regulation of the tissue-specific expression of transferrin gene.

Transferrin (Tf), the plasma protein involved in iron transport, seems to play complex physiological roles related to cell function, differentiation and proliferation. The protein is essentially synthesized in hepatocytes, but also in Sertoli cells, in the epithelial cells of the choroid plexus in rodents and in oligodendrocytes in all species analyzed. In this manuscript, we review the results obtained on Tf gene expression in the different cellular systems in which the protein is synthesized. In vitro and ex vivo experiments indicate that different combinations of transcription factors are necessary in different subsets of cells to achieve Tf tissue-specific expression. Several lines of transgenic mice were generated in which the expression of reporter genes is under the control of different Tf regulatory regions. More recently, transgenic mice were obtained using the complete human Tf gene and its 5' and 3' flanking sequences. These mice constitute the first model in which the physiological consequences of a specific Tf over expression in oligodendrocytes can be studied. Although much information is now available, further work is still necessary for a full understanding of the in vivo mechanisms responsible for the regulation of Tf gene expression.

Animals↗

PubMedDate not supplied

Extracellular signal-regulated kinases (ERK) 1, 2 are required for luteinizing hormone (LH)-induced steroidogenesis in primary Leydig cells and control steroidogenic acute regulatory (StAR) expression.

The luteinizing hormone (LH) plays a critical role in steroidogenesis, by stimulating cAMP-dependent protein kinase A (PKA) and phospholipase A2 activity, and by mobilizing calcium and chloride ions. In contrast, whether the ERK 1, 2 mitogen-activated protein (MAP) kinases are involved in LH-induced steroidogenesis is less obvious. Here, we sought to clarify this point in rat primary Leydig cells, naturally bearing the LH receptor (LH-R) in male, and in the mouse tumoral Leydig cell line (MLTC 1). Pre-incubation of both cell types with the mitogen-activated protein kinase kinase (MEK) inhibitors U0126 and PD98059 reduced LH-induced steroidogenesis, and tonically enhanced the expression of the StAR protein. Furthermore, ERK1, 2 were inducibly phosphorylated following LH exposure of MLTC 1 cells. Altogether, our results indicate that in primary as well as in tumoral Leydig cells, inhibiting MEK dampened LH-induced steroidogenesis but enhanced basal as well as LH-induced StAR expression, suggesting that ERK1,2 could be involved in these responses.

Animals↗