IRE1α通过UPR影响非洲爪蟾胚胎发育

摘要

Objective: To explore whether IREloi can mediate unfolded protein response (UPR) during embryo development of xenopus laevis and rescue the developmental defects induced with tunicamycin. Methods; IRE la mRNA or specific antisense oligonucleotids morpholino were microinjected to the 4 blastomeres of 4-cell stage embyos to overexpress or knockdown gene expression; the splicing of XBP1 was tested with RT-PCR to reflect the UPR;Tunicamycin was used to treat the embryos to induce the endo-plasmic reticulum stress (ERS). Results; The splicing of XBP1 was increased when IRE la was overexpressed while the splicing was decreased when IRE la was knockdwon. Embryos developmental defects and the increase of XBP1 splicing were induced with tunicamycin while knockdown of XBP-1 partially rescued the defects and knockdown of IRE la rescued the defects significantly and the increased XBP1 splicing. Conclusion; IRE la can mediate UPR during xenopus development. Knockdown of IRE la rescued the developmental defects induced with TM and partially by the IREla/XBPl pathway.%目的:探讨IRE1α在非洲爪蟾胚胎发育中是否介导未折叠蛋白反应(UPR),并在衣霉素(tunicamycin,TM)诱导的胚胎发育畸形中发挥挽救作用.方法:利用TM处理胚胎激发内质网应激；应用显微注射mRNA方法实现基因过表达,通过注射特异的反义寡核苷酸morpholino(MO)实现基因封闭;RT-PCR检测XBP1剪切情况.结果:XBP1剪切随IRE1α过表达及封闭而增加或减少:TM处理导致胚胎发育畸形,xBP1剪切增加,封闭XBPI可部分挽救发育畸形；封闭IRE1a可明显挽救发育畸形,XBP1剪切恢复.结论:IRE1α在非洲爪蟾胚胎发育中介导UPR,封闭IRE1α可逆转TM诱导的胚胎发育畸形,部分通过XBP1途径发挥作用.