黄脸油葫芦精巢TeHsc70和TeHsp90敲低对20E相关基因表达的影响

Effects of Hsc70 and Hsp90-RNAi on expression of 20E-related genes in testis of Teleogryllus emma

为了研究Hsc70和Hsp90与ERR和20E下游初、次级应答基因的关联性,明确Hsc70和Hsp90如何参与20E信号通路进而调控雄性生殖,以雄性成体黄脸油葫芦(Teleogryllus emma Ohmachi&Matsuura)为实验对象,设实验组、阴性对照组和Control组。实验组注射ds TeHsc70和ds TeHsp90;阴性对照组注射EGFP-dsRNA,干扰48、72和96h后qPCR检测RNA干扰(RNA interference,RNAi)效率和20E相关基因的表达。结果显示,TeEcR、TeERR和TeUsp在空白组成体精巢中高表达,并且TeERR mRNA的表达显著高于TeEcR和TeUsp(P<0.05)。ds TeHsc70+ds TeHsp90干扰96h后,TeHsc70和TeHsp90 mRNA的表达显著下调(P<0.05),并且TeERR和20E初、次级应答基因TeEcR、TeUsp、TeE75B和TeHR3mRNA的表达也随之极显著下调(P<0.01)。据此推测,TeHsc70、TeHsp90和TeERR均参与20E信号通路,并且TeHsc70、TeHsp90和TeERR对于20E信号通路维持精巢的发育和形态建成方面必不可少。

To explore the relationship between HscTO and Hsp90 with ERR, 20E primary and sec- ondary response genes, and how do the ERR and 20E-related genes involved in the 20E signaling pathway and male fertility, adult male crickets Teleogryllus emma Ohmschi Matsumura were divided into the experimental group, negative group, and control group. Crickets of the experi- mental group were injected with dsTeHscTO and dsTeHsp90, and the negative group received EGFP-dsRNA, then the efficiency of TeHscTO- and TeHspgO-RNAi and expression of TeERR and 20E-related genes were detected in the experimental group after the crickets treated for 48, 72 and 96 h. Results showed that the TeEcR, TeERR, and TeUsp were highly expressed in free-treated adult testes, and TeERR expression was significantly higher than TeEcR and TeUsp （P〈 0.05）. In addition, expressions of the TeHsc70 and TeHsp90 mRNA is significantly down-regu- lated（P〈0.05） after the crickets treated for 96 h using 2 000 ng dsTeHsc7Oq-dsTeHsp90. Fur- thermore, expression of the TeERR and 20E primary and secondary response genes, TeEcR, TeUsp, TeE75B, and TeHR3 mRNA, could also be significantly down-regulated （P〈0.01）. All these results indicate that Hsc70 and Hsp90 are involved in the 20E signal transduction path- way and maintain the development and morphogenesis of testis in crickets T. emma.