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Mol. Hum. Reprod. Advance Access published online on March 23, 2006
Molecular Human Reproduction, doi:10.1093/molehr/gah247
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© The Author 2006. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org
Received August 25, 2005
Accepted November 11, 2005

Article

Transcriptome analysis of FSH and FSH variant stimulation in granulosa cells from IVM patients reveals novel regulated genes

S. Perlman 1, T. Bouquin 2 *, B.van den Hazel 3, T.H. Jensen 4, H.T. Schambye 5, S. Knudsen 4, and J.S. Okkels 2

1 Maxygen, Hørsholm; Department of Gynecology and Obstetrics, Rigshospitalet, Blegdamsvej, Copenhagen
2 Maxygen, Hørsholm
3 Maxygen, Hørsholm; Present address: Høiberg A/S, Store Kongensgade 59A, DK-1264 Copenhagen K, Denmark
4 The Center for Biological Sequence Analysis, Technical University of Denmark, Lyngby, Denmark
5 Maxygen, Hørsholm; Present address: Gastrotech Pharma A/S - Nyhavn 43B, DK-1051 Copenhagen K, Denmark

* To whom correspondence should be addressed.
T. Bouquin, E-mail: TBo{at}maxygen.dk


  Abstract

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.

Keywords: FSH/gene regulation/granulosa cells/IVM patients/microarray.
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