Epigenetic and transcriptional regulation of gonadotropin induced gene expression in ovarian granulosa cells

From the RNA seq analysis of ovarian granulosa cells (GCs) we got a clue that FSH-induced epigenetic and/or transcriptional regulation is essential for LH effects on the GC genes. We further investigated the mechanism of gonadotropin-induced gene repression or gene induction in GCs by methyl-sequencing of genomic DNA. GCs were isolated from the 4-wk-old wildtype rat ovaries before gonadotropin administration (Basal), 48h after PMSG administration (PMSG), and 4hr after hCG administration following PMSG-priming (4h hCG). Genomic DNA was extracted from the GCs and o approximately 200 ng of gemomic DNA was used for each methyl-seq library. We used four replicates for each group and each replicate is made of GCs from 3 rats. The genomic DNA was sheared and the library prepared using the NEBNext Methyl-seq kit from New England Biolabs, and the library was constructed by Novogene Coporation.