Mol. Hum. Reprod. Advance Access originally published online on March 23, 2006
Molecular Human Reproduction 2006 12(3):135-144; doi:10.1093/molehr/gah247
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Transcriptome analysis of FSH and FSH variant stimulation in granulosa cells from IVM patients reveals novel regulated genes
1Maxygen, Hørsholm, 2Department of Gynecology and Obstetrics, Rigshospitalet, Blegdamsvej, Copenhagen and 3The Center for Biological Sequence Analysis, Technical University of Denmark, Lyngby, Denmark
4 To whom correspondence should be addressed at: Maxygen, Agern Allé 1, DK-2970 Hørsholm, Denmark. E-mail: TBo{at}maxygen.dk
5 Present address: Høiberg A/S, Store Kongensgade 59A, DK-1264 Copenhagen K, Denmark
6 Present address: Gastrotech Pharma A/S - Nyhavn 43B, DK-1051 Copenhagen K, Denmark
FSH is crucial for oocyte maturation and fertility and is the main component in infertility treatment in assisted reproduction. The granulosa cells expressing the FSH receptor interact with the oocyte and provide nourishing substrates controlling the oocyte maturation. Thus, transcriptome analysis of granulosa cells stimulated by FSH is of major importance in understanding the communication between oocytes and granulosa cells. In this study, gene expression profiles were assessed in human granulosa cells from normal cycling in vitro maturation (IVM) patients using oligonucleotide gene chips. Granulosa cells were stimulated for 2 h with either FSH or a previously generated glycosylated FSH variant (FSH1208) that exhibited increased in vivo activity because of prolonged half-life. The analysis identified 74 significantly FSH/FSH1208 regulated genes. Amongst these were well known FSH regulated genes as well as genes not previously described to be important in the FSH signalling pathway. These novel FSH regulated genes include transcription factors [cAMP responsive element modulator (CREM)/inducible cAMP early repressors (ICER), GATA 6, ZFN 361, Bcl11a, CITED1 and TCF 8] and other regulatory proteins and enzymes (IGF-BP3, syntaxin and PCK1) possibly important for oocyte/granulosa cell interaction and function. Array data were validated for 13 genes by northern blots or RT–PCR. Furthermore, no significant differences in gene regulation were detected between the two FSH analogs. This work uncovers novel data important for understanding the folliculogenesis. Furthermore, the results suggest that FSH1208 has a gene expression profile like FSH and thus, in the light of known prolonged in vivo activity, might be a candidate for improved infertility treatment.
Key words: FSH/gene regulation/granulosa cells/IVM patients/microarray
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