Investigation of the mechanisms of progesterone protection following oxygen-glucose deprivation in organotypic hippocampal slice cultures

Abstract

This study aimed to test the hypothesis that progesterone is neuroprotective against oxygen-glucose deprivation (OGD) through its conversion to the active metabolite allopregnanolone (AlloP) and the potentiation of GABA(A) receptors. Organotypic hippocampal cultures were exposed to 2h of OGD and the resulting cell death was quantified 24h later using combined propidium iodide and Hoechst immunostaining. Initially, we confirmed, that both progesterone and AlloP were protective in terms of reducing cell death following OGD in hippocampal cultures and for both, the optimal level of protection was observed at a concentration of 0.1μM. However, the protective effect of progesterone was absent in the presence of finasteride (10μM) which inhibits the metabolism of progesterone to active metabolites, including AlloP. In addition, the concurrent application of picrotoxin (100μM), a potent GABA(A) receptor antagonist, prevented the protection previously seen by either progesterone or AlloP alone. These results indicate that progesterone protects hippocampal cultures from cell death following OGD largely due to its conversion to AlloP and that GABA(A) receptors are important mediators of the protective effects of both progesterone and AlloP.