Identification of differential gene expression in in vitro FSH treated pig granulosa cells using suppression subtractive hybridization.
FSH, which binds to specific receptors on granulosa cells in mammals, plays a key role in folliculogenesis. Its biological activity involves stimulation of intercellular communication and upregulation of steroidogenesis, but the entire spectrum of the genes regulated by FSH has yet to be fully characterized. In order to find new regulated transcripts, however rare, we have used a Suppression Subtractive Hybridization approach (SSH) on pig granulosa cells in primary culture treated or not with FSH. Two SSH libraries were generated and 76 clones were sequenced after selection by differential screening. Sixty four different sequences were identified, including 3 novel sequences. Experiments demonstrated the presence of 25 regulated transcripts.A gene ontology analysis of these 25 genes revealed (1) catalytic; (2) transport; (3) signal transducer; (4) binding; (5) anti-oxidant and (6) structural activities. These findings may deepen our understanding of FSH's effects. Particularly, they suggest that FSH is involved in the modulation of peroxidase activity and remodelling of chromatin.
Bonnet
A
A
INRA laboratoire de Génétique cellulaire, BP52627 chemin de borde rouge, 31326 Castanet cedex, France. abonnet@toulouse.inra.fr
Frappart
P O
PO
Dehais
P
P
Tosser-Klopp
G
G
Hatey
F
F
D000818:Animals; D015152:Blotting, Northern; D002478:Cells, Cultured / Q000187:drug effects / Q000378:metabolism; D002874:Chromosome Mapping; D018076:DNA, Complementary / Q000235:genetics; D005260:Female; D005640:Follicle Stimulating Hormone / Q000494:pharmacology*; D020869:Gene Expression Profiling / Q000379:methods*; D005786:Gene Expression Regulation; D015723:Gene Library; D006107:Granulosa Cells / Q000187:drug effects* / Q000378:metabolism; D009693:Nucleic Acid Hybridization; D011962:Receptors, FSH / Q000187:drug effects / Q000502:physiology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D013382:Subtraction Technique; D034421:Sus scrofa; D014158:Transcription, Genetic
D018076:DNA, Complementary; D011962:Receptors, FSH; D005640:Follicle Stimulating Hormone