Optimization of pAkt Expression and Detection in Bovine Granulosa Cells

Sarah M. Piet, University of New Hampshire
David H. Townson PhD, University of Vermont
Paul C. Tsan PhD, University of New Hampshire

Abstract

The cellular mechanisms that regulate the fate of granulosa cells in the bovine follicle remain relatively unknown. One pathway believed to be involved in regulation is the protein kinase B (pAkt) pathway. In general, the pAkt proteins are intracellular kinases that are thought to have a role in cell survival, preventing apoptosis, and cell proliferation. However in the context of bovine granulosa cells (bGCs) the conditions that lead to upregulation of pAKt are not entirely clear, nor is the effect of pAkt. In the present study we tested the conditions and procedures for cell culture and immunodetection which would optimize the protein expression and detection of pAkt in bGCs. Future work will focus on determining if pAkt signaling influences granulosa cell resistance to immune-mediated apoptosis.