中文摘要：

体外培养的奶牛，绵羊和小鼠的附植前胚胎在热应激条件下能够合成热休克蛋白70以增强胚胎的耐热性保护其不受热损伤。而钙调蛋白（CaM）是细胞内最重要的Ca2＋受体蛋白并参与细胞活动中重要基因的转录活动。本实验旨在研究CaM参与小鼠（Mus muscttlus）胚胎热应激诱导型热休克蛋白70（HSP70）的表达，且明确CaM参与HSP70表达的具体途径。将发育至早期囊胚的胚胎分成空白对照（37℃）组、W7处理非热应激（370C＋W7）组、热应激（39℃）组和W7处理热应激（39℃＋w7）组。提取总RNA并分别检测HSP70、如M和热休克转录因子1（HSF1）mRNA表达；提取胞浆蛋白用Western blot技术检测HSP70、CaM和HSF1表达；用免疫共沉淀法检测HSP70-CaM和HSP70-HSF1复合物。结果发现，W7能显著抑制39℃热应激1h小鼠胚胎HSP70mRNA和蛋白的表达（P〈0．05）；W7对其他各组胚胎HSF1，mRNA表达的影响不显著（P〉0．05），但能显著降低39℃热应激1h小鼠胚胎HSF1蛋白表达（P〈0．05）；39℃热应激时，胚胎HSP70．CaM复合物增多，HSP70-HSF1复合物减少。本研究表明，小鼠胚胎受热应激时，CaM通过与HSP70竞争性结合，解离出HSF1，从而使HSP70大量表达。本研究揭示了CaM参与小鼠胚胎热激反应中HSP70表达的一种途径。可为研究胚胎耐热性以及热激信号转导提供基础资料。

英文摘要：

Heat shock protein（HSP）70 is synthesized in the preimplantation embryos of cow（Bos tarurs）, ewe （Ovis aries） and mice（Mus musculus） under the conditions of heat shock in vitro, which enhances theirthermotolerance to protect them from further heat damage. And Calmodulin（CaM）is the one of most important multi functional receptor of Ca2＋ and plays a key role in regulating the important gene expression in cellactivities. To clarity whether the CaM participating in the expression of HSP70 in the mice embryos with heat shock and verify its mechanism, mice embryos were cultured in vitro to the blastocyst stage and then wererandomly divided into control group（37 ℃）, group treated with W7 without heat shock（37 ℃ ＋W7）, heat shock group（39℃ ）and heat shock group treated with W7. The method of RT-PCR was used to detect the geneexpression of CaM, HSP70 and heat shock factorl（HSF1）, and the method of Westem blot was used to detect the expression of CaM, HSP70 and HSF1. The combinations of both HSP70-CaM and HSP70-HSF1 weredetected by using co-immunoprecipitation（Co-IP）. The results showed that the CaM and HSP70 mRNA expression significantly increased in the mice embryos treated with 39℃ for 1 h （P〈0.05）. However, the CaMand HSP70 mRNA expression significantly decreased in both groups of embryo cultured at 37℃ （P〈0.05） and 39℃ （P〈0.01） treated with W7; W7 had no effect on the HSF1 mRNA expression in any groups of embryo.The synthesis of CaM, HSP70 and HSF1 significantly increased in the embryos treated at 39℃ for 1 h （P〈 0.05）, and the CaM synthesis significantly decreased in the embryos cultured both at 37℃ and 39℃ andtreated with W7 （P〈0.05）. W7 had no effect on neither HSP70 nor HSF1 synthesis in the embryos cultured at 37℃ （P〉0.05） while it inhibited both HSP70 and HSF1 synthesis in the embryos cultured at 39℃ （P〈0.05）.The study also found that CaM combined with HSP70 and formed CaM-HSP70 complex in the mice embryos.