Immune-labeling of cytochrome P450scc and its associate electron transferring enzymes in mouse ovary tissue-sections

Background Steroid biosynthesis is mediated by cytochrome P450scc enzyme, a rate-limiting step involved in the conversion of cholesterol to pregnenolone in gonads. The cytochrome P450scc enzyme is accompanied by electron transferring enzyme adrenodoxin and adrenodoxin reductase, all present on mitochondrial integral membrane. This research article basically reveals the sites of expression of cytochrome P450scc, adrenodoxin, and adrenodoxin reductase in mouse ovarian system using high-resolution microscopy. The study has covered thorough expression of these mitochondrial systems in different phases of follicle formation and surrounding interstitial stromal cells. Methods We stained mouse ovary sections at various stages of their folliculogenesis using antisera generated against cytochrome P450scc, Adrenodoxin, and Adrenodoxin reductase enzymes. We also stained ovary sections with phalloidin conjugates 488 and WGA-Lectin conjugates 555 to identify different stages of folliculogenesis/ovulation cycle. Results The current study showed staining for lectin binding moieties and phalloidin on the zona pellucida region of early-preantral and antral follicles. We identified the expression of P450scc, adrenodoxin, and adrenodoxin reductase enzymes in thecal cells of primordial follicles, primary follicles, secondary follicles, and interstitial stromal cells. Conclusion The information generated here might be useful in identifying specific role of cytochrome P450scc expressing cells in Addison's disease and autoimmune primary ovarian insufficiency.