PD35-12 KDM5A, A HISTONE H3K4 DEMETHYLASE, REGULATES SPERMATOGENESIS VIA ITS ACTION ON GERM CELL

You have accessJournal of UrologyStem Cell Research1 Apr 2016PD35-12 KDM5A, A HISTONE H3K4 DEMETHYLASE, REGULATES SPERMATOGENESIS VIA ITS ACTION ON GERM CELL Hidenori Nishio, Kentaro Mizuno, Moritoki Yoshinobu, Hideyuki Kamisawa, Akihiro Nakane, Satoshi Kurokawa, Tetsuji Maruyama, Yutaro Hayashi, and Takahiro Yasui Hidenori NishioHidenori Nishio More articles by this author , Kentaro MizunoKentaro Mizuno More articles by this author , Moritoki YoshinobuMoritoki Yoshinobu More articles by this author , Hideyuki KamisawaHideyuki Kamisawa More articles by this author , Akihiro NakaneAkihiro Nakane More articles by this author , Satoshi KurokawaSatoshi Kurokawa More articles by this author , Tetsuji MaruyamaTetsuji Maruyama More articles by this author , Yutaro HayashiYutaro Hayashi More articles by this author , and Takahiro YasuiTakahiro Yasui More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2016.02.1084AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES The attenuation of spermatogonial stem cell activity in cryptorchid testes alters spermatogenesis and affects fertility. We have shown that lysine (K)-specific demethylase 5A (Kdm5a) is more highly expressed in undescended testes than in normal testes of juvenile rats, suggesting that Kdm5a regulates spermatogenesis via histone H3K4 modification (J Urol., 2014). In the present study, we aimed to elucidate the function of Kdm5a, a key epigenetic regulator, in the testes in vitro. METHODS We prepared GC-1 spg, a mouse spermatogonial cell line; TM4, a mouse Sertoli cell line; and TM3, a mouse Leydig cell line. These cells were transfected with pEZ-M03 (GeneCopoeiaTM) as control, the Gfp expression vector, or the Kdm5a-Gfp expression vector. The cells were harvested 24 h after transfection for mRNA and protein extraction. We assessed the expression of Kdm5a and genes related to spermatogenesis (Esr1, Esr2, Gdnf, Gfra1, Kit, Klf4, Myc, Nanog, Pgr, Pou5f1, Ret, and Thy1) by using quantitative reverse transcription polymerase chain reaction (RT-PCR). Moreover, we assessed the H3K4 modification state by using western blot analysis and the expressions of KDM5A and the spermatogenesis-related genes by using RT-PCR. RESULTS The Kdm5a and H3K4me2/me3 expression levels were significantly higher in GC-1, TM4, and TM3 when transfected with the Kdm5a expression vector. Kdm5a overexpression in the GC-1 cells led to increased expression levels of Esr2, Neurog3, Pou5f1, Ret, and Thy1, although Kdm5a overexpression in the TM4 and TM3 cells did not change the expression levels of these genes. CONCLUSIONS We reported Kdm5a expressed in the germ cells of the testes of juvenile rats (J Urol., 2014). Although the testes consist of germ, Sertoli, and Leydig cells, GC-1 cells, not TM4 and TM3, changed the expression levels of the spermatogenesis-related genes through Kdm5a overexpression. This suggests that spermatogenesis is regulated by the action of Kdm5a on germ cells. © 2016FiguresReferencesRelatedDetails Volume 195Issue 4SApril 2016Page: e848 Advertisement Copyright & Permissions© 2016MetricsAuthor Information Hidenori Nishio More articles by this author Kentaro Mizuno More articles by this author Moritoki Yoshinobu More articles by this author Hideyuki Kamisawa More articles by this author Akihiro Nakane More articles by this author Satoshi Kurokawa More articles by this author Tetsuji Maruyama More articles by this author Yutaro Hayashi More articles by this author Takahiro Yasui More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...