MP66-08 CLUSTERIN PRODUCED BY SERTOLI CELLS INHIBITS HEAT STRESS-INDUCED APOPTOSIS IN RAT TESTIS

Abstract

You have accessJournal of UrologyInfertility: Basic Research, Physiology & Pathophysiology1 Apr 2014MP66-08 CLUSTERIN PRODUCED BY SERTOLI CELLS INHIBITS HEAT STRESS-INDUCED APOPTOSIS IN RAT TESTIS Kei Matsushita, Hideaki Miyake, and Masato Fujisawa Kei MatsushitaKei Matsushita 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.2014.02.2055AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES The objectives of this study were to characterize the changes in expression of clusterin that is shown to have a chaperon-like activity and act as a powerful inhibitor against a wide variety of proapoptotic stimuli, in rat testis following heat stress, and to investigate whether the inhibition of clusterin expression in rat Sertoli cells enhances apoptosis induced by heat exposure. METHODS Adult male Sprague-Dawley rats were used in this study. The scrotum of rats were completely immersed in a water bath and exposed to either 43C or 37C as a control for 15 minutes. Rats were then sacrificed on days 1, 4, 7, 14 and 21 after heat treatment. Time-course changes in testicular weight and germ cell number were measured, expression levels of clusterin mRNA and protein were evaluated respectively, and immunohistochemical staining of clusterin and TUNEL assay were also performed. We then purified Sertoli cells from rat testes, and expression vector containing short hairpin RNA (shRNA) targeting rat clusterin gene or control vector alone was transiently transfected into Sertoli cells. Following exposure to heat stress at 41C for 12 hours, changes in clusterin mRNA expression and apoptotic index were analyzed by real-time RT-PCR and TUNEL assay, respectively. RESULTS Both testicular weight and germ cell number in rat testis after heat treatment were markedly reduced in time-dependent manners compared with those after control treatment. To the contrary, expression levels of clusterin mRNA and protein were significantly up-regulated following heat treatment and reached to the highest levels on day 21, while the apoptotic index was highly increased 1 day after heat treatment, and was maintained at higher levels than that after control treatment. In cultured rat Sertoli cells exposed to heat stress, clusterin mRNA was markedly up-regulated in a time-dependent manner after transfection with control vector; however, shRNA targeting clusterin resulted in the inhibition of clusterin mRNA expression by more than 70%. Furthermore, apoptotic index in these Sertoli cells after treatment with shRNA targeting clusterin was significantly higher than that after treatment with control vector, and this difference was most prominent within 24 hours after heat stress. CONCLUSIONS These findings suggest that clusterin secreted by Sertoli cells protect testis from injury induced by heat stress; accordingly, further up-regulation of clusterin expression in the testis following the exposure to heat stress could be a promising approach for the inhibition of heat-induced disruption of spermatogenesis. © 2014FiguresReferencesRelatedDetails Volume 191Issue 4SApril 2014Page: e743 Advertisement Copyright & Permissions© 2014MetricsAuthor Information Kei Matsushita 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 ...