MP66-07 NEPHRIN-PODOCIN COMPLEX PLAY AN IMPORTANT ROLE IN SPERMATOGENESIS

Abstract

You have accessJournal of UrologyInfertility: Basic Research, Physiology & Pathophysiology1 Apr 2014MP66-07 NEPHRIN-PODOCIN COMPLEX PLAY AN IMPORTANT ROLE IN SPERMATOGENESIS Noritoshi Enatsu, Teruo Fukuda, Keisuke Okada, Kei Matsushita, Koji Chiba, Hideaki Miyake, and Masato Fujisawa Noritoshi EnatsuNoritoshi Enatsu More articles by this author , Teruo FukudaTeruo Fukuda More articles by this author , Keisuke OkadaKeisuke Okada More articles by this author , Kei MatsushitaKei Matsushita More articles by this author , Koji ChibaKoji Chiba More articles by this author , Hideaki MiyakeHideaki Miyake More articles by this author , and Masato FujisawaMasato Fujisawa More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2014.02.2054AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Nephrin and podcin are well known proteins which are connected each other and form slit diaphragm in the kidney podocyte. Recently, podocin is also detected in the testis, but extrarenel nephrin expression has not been confirmed. So we firstly aim to distinguish the expression of nephrin and analyze the relationship between nephrin and podocin in rat testis. We secondary investigate the expression patterns of nephrin and podocin in human testis with relation to Johnsen□fs socore. METHODS By using 16 weeks old Sprague-Dawley rat, the expression and the localization of nephrin was analyzed by quantitative RT-PCR, Western blot analysis and immunofluorescence. As the testicular injury group, cisplatin was administerd at a single injection of 4mg/kg and 7mg/kg intraperitoneally. We then analyzed the expression and localization of testicular nephrin and podocin using human samples. Human testicular tissues were collected from 28 patients who underwent testicular sperm extraction at Kobe University Hospital. Written informed consent to perform this study was obtained from each of the included patients, and the study design was approved by the Research Ethics Committee of our institution. Based on the pathological diagnosis, samples were divided into two groups; 8 samples of obstructive azoospermia (OA) and 20 samples of non-obstructive azoospermia (NOA). The expression levels of nephrin were determined by RT-PCR and the localization was analyzed by immunofluorescent staining. RESULTS We certainly detected the expression of nephrin gene and protein in rat testis. Immunofluorescence staining revealed the co-localization of nephrin and podocin near the basal membrane of seminiferous tubules and cell membranes of Sertoli cells. Protein complex immunoprecipitation revealed that nephrin and podocin bind to each other. After cisplatin administration, the amount of the gene and protein of nephrin significantly decreased. In human samples, expression and co-localization of nephrin and podocin were also confirmed. The expression level of nephrin was significantly higher in OA compared to NOA. The staining pattern of nephrin also sifted from linear to granular according to deteriorate the Johnsen score. CONCLUSIONS Nephrin and podocin are co-expressed near basal membrane of seminiferous tubules in testis. It suggests that nephrin-podocin complex may play an important role in spermatogenesis as a component of junction between Sertoli cells and germ cells. To support this fact, the expression of nephrin and podocin was low and irregular in patients of hypospermatogenesis and sertoli cell only syndrome. © 2014FiguresReferencesRelatedDetails Volume 191Issue 4SApril 2014Page: e743 Advertisement Copyright & Permissions© 2014MetricsAuthor Information Noritoshi Enatsu More articles by this author Teruo Fukuda More articles by this author Keisuke Okada More articles by this author Kei Matsushita More articles by this author Koji Chiba More articles by this author Hideaki Miyake More articles by this author Masato Fujisawa More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...