Global Profiling of Activin Target Genes in Mouse Granulosa Cells

In order to understand the mechanisms and pathways by which activin regulates follicle function, we performed a microarray study and identified activin regulated genes in mouse ovarian granulose cells. Primary cultured ovarian granulosa cells collected from day 21-23 old immature mice were treated with Activin A (A), Activin A plus its antagonist follistatin (AplusF), and PBS (P, vehicle) for 24 h. RNA from each treatment group was isolated and the experiments were repeated four times. Samples from experiment one were assigned as P1, A1 and A+F1, samples from experiment two as P2, A2 and A+F2, and so on. About 100 ng of each RNA sample was amplified using a modified Eberwine T7-based amplification protocol and hybridized to the Illumina Sentrix Mouse-6 Expression BeadChip (Illumina, Inc., San Diego, CA) that contains 48,000 sets of oligonucleotides.