813 FUNCTIONAL ANALYSIS OF RHO SIGNALING CASCADE CONCERNED WITH CELL ADHESION IN THE PROCESS OF TESTICULAR DEVELOPMENT

Abstract

You have accessJournal of UrologySexual Function/Dysfunction/Andrology: Basic Research (I)1 Apr 2013813 FUNCTIONAL ANALYSIS OF RHO SIGNALING CASCADE CONCERNED WITH CELL ADHESION IN THE PROCESS OF TESTICULAR DEVELOPMENT Kentaro Mizuno, Yutaro Hayashi, Hideyuki Kamisawa, Yoshinobu Moritoki, Hidenori Nishio, Makoto Imura, Yasuhiro Shibata, Satoshi Kurokawa, Akihiro Nakane, Toshiki Kato, Tetsuji Maruyama, Yoshiyuki Kojima, and Kenjiro Kohri Kentaro MizunoKentaro Mizuno Nagoya, Japan More articles by this author , Yutaro HayashiYutaro Hayashi Nagoya, Japan More articles by this author , Hideyuki KamisawaHideyuki Kamisawa Nagoya, Japan More articles by this author , Yoshinobu MoritokiYoshinobu Moritoki Nagoya, Japan More articles by this author , Hidenori NishioHidenori Nishio Nagoya, Japan More articles by this author , Makoto ImuraMakoto Imura Nagoya, Japan More articles by this author , Yasuhiro ShibataYasuhiro Shibata Nagoya, Japan More articles by this author , Satoshi KurokawaSatoshi Kurokawa Nagoya, Japan More articles by this author , Akihiro NakaneAkihiro Nakane Nagoya, Japan More articles by this author , Toshiki KatoToshiki Kato Nagoya, Japan More articles by this author , Tetsuji MaruyamaTetsuji Maruyama Nagoya, Japan More articles by this author , Yoshiyuki KojimaYoshiyuki Kojima Fukushima, Japan More articles by this author , and Kenjiro KohriKenjiro Kohri Nagoya, Japan More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2013.02.379AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES During early stage of testis development, formation of seminiferous tubule is preceded with adhesion between germ cells and somatic cells. However, the relationship between the mechanisms of testis development and cell adhesion remains unclear. Previously, we detected Rock1 gene as testis development related gene, and Rock1 gene is belong to Rho signaling cascade which regulate cell adhesion. In this study, to clarify the role of cell adhesion in the process of testis development comprehensively, we performed functional analysis of factors concerned with Rho signaling cascade in testis. METHODS From testicular tissue samples of patients with 46,XX testicular DSD (XX testis) and age-matched patients with hydrocele testis (XY testis), mRNA was extracted and cDNA libraries were obtained, respectively. We explored the genes associated with differential expression between XY and XX testis using by PCR-subtractive hybridization methods. Subsequently, we detected that ROCK1 gene is up-regulated in XX testis without SRY gene. We investigated distribution of not only Rock1 protein but also β1-integrin and Rho proteins which are upstream of Rho signaling cascade in testicular tissues of human and experimental animals. Furthermore, to analyze the function of Rock1 gene, Y-27632, selective inhibitor of Rock1, was added into cultured Sertoli cells and expression changes of each gene including Rho signaling cascade were examined by RT-PCR. This study was approved by the institutional review board in our hospital. RESULTS ROCK1 protein was detected in spermatogonia and Sertoli cells in immature testis of human. While, in adult mice, Rock1 protein was detected in spermatocytes, spermatids and Leydig cells in seminiferous tubule stage-specific manner. β1-integrin protein was detected in germ cells as well as Rock1 protein. In addition of Y-27632 into cultured Sertoli cells, expressions of Sox9 gene were significantly decreased, but expression changes of RhoA and RhoB gene, which is upstream of Rock1 gene, were not significant. CONCLUSIONS Rock1 protein was expressed in both germ cells and somatic cells, suggesting that Rock1 is associated with cell adhesion in each other. Furthermore, we elucidated that Sox9 gene is up-regulated by Rock1 gene and regulated by several signaling pathway. Since it is reported that niche surrounding spermatogonial stem cells is impaired in knockout mice of Sox9 gene, it is suggested that Rho signaling cascade upregulates Sox9 gene and is concerned with testis development through adhesion between germ cells and somatic cells. © 2013 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 189Issue 4SApril 2013Page: e334 Advertisement Copyright & Permissions© 2013 by American Urological Association Education and Research, Inc.MetricsAuthor Information Kentaro Mizuno Nagoya, Japan More articles by this author Yutaro Hayashi Nagoya, Japan More articles by this author Hideyuki Kamisawa Nagoya, Japan More articles by this author Yoshinobu Moritoki Nagoya, Japan More articles by this author Hidenori Nishio Nagoya, Japan More articles by this author Makoto Imura Nagoya, Japan More articles by this author Yasuhiro Shibata Nagoya, Japan More articles by this author Satoshi Kurokawa Nagoya, Japan More articles by this author Akihiro Nakane Nagoya, Japan More articles by this author Toshiki Kato Nagoya, Japan More articles by this author Tetsuji Maruyama Nagoya, Japan More articles by this author Yoshiyuki Kojima Fukushima, Japan More articles by this author Kenjiro Kohri Nagoya, Japan More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...