Comparison of POU۵F۱ Expression with Three DifferentAntibodies in Spermatogonia S tem Cell Populationsof Neonatal and Adult Mice

Objective: Spermatogenesis is a complex and elaborate differentiationprocess and is vital for male fertility. Currently, manygenes have been identified that affect the proper functioning ofdifferentiation during spermatogenesis. POU۵F۱ (POU class ۵homeobox ۱) is a crucial transcription factor for pluripotencyand self-renewal, thus playing a vital role in accurate spermatogenesis.Materials and Methods: In this experimental s tudy, we examinePOU۵F۱ protein localization by three various kinds of antibodiesin the neonatal and adult mice tes tis section and isolatedspermatogonial s tem cells (SSCs) by immunohis tochemis try(IHC) and immunocytochemis try (ICC). Also, we examine themRNA expression of POU۵F۱ in neonatal and adult mice SSCsby Fluidigm real-time polymerase chain reaction. The POU۵F۱protein-protein network was cons tructed and gene enrichmentanalysis was performed.Results: IHC and ICC data by all three antibodies showed thatPOU۵F۱ expression was much higher in neonate mice than inadult mice. Fluidigm analysis represented significantly higherexpression (P value < ۰.۰۵) of POU۵F۱ in the adult tes tis incomparison to the neonate and a significant expression (P value< ۰.۰۵) level of germ cells gene POU۵F۱ in neonate SSCs (۱-۲week) than ۱۶–۲۴-week SSCs. In silico data also show a s trongassociation of POU۵F۱ with other markers involved in spermatogenesis.Conclusion: These results indicate that POU۵F۱ is a necessarytranscription factor of tes ticular germ cells that can be supportivefor the analysis of germ cells development both in vitro andin vivo.

کلیدواژه ها:

Immunocytochemis try ، Immunohis tochemis try ، POU۵F۱ ، Spermatogenesis ، Spermatogonial S tem Cells (SSCs)