中文摘要：

本试验旨在研究冷应激对猪卵母细胞发育能力及冷诱导RNA结合蛋白（CIRP）基因表达的影响。常规体外成熟猪GV期卵母细胞,置于室温（25℃）下冷应激处理0.5、1、2、6、12和24 h,观察冷应激处理对其后卵母细胞成熟率和体外受精胚胎发育率的影响和微丝、微管等亚细胞结构的变化,Real-time PCR检测猪卵母细胞CIRP mRNA的表达。结果表明：短时间（〈2 h）冷应激处理后各组成熟率和体外受精卵裂率均与对照组无异（P〉0.05）;微丝、微管及染色体等亚细胞结构均无明显改变（P〉0.05）,但2 h组的囊胚发育率较对照组有明显下降（9.7%对12.3%,P〈0.05）。冷应激处理超过2 h后,各组成熟率和囊胚发育率与对照组相比均显著降低（P〈0.05）,微丝、微管正常率均较对照组显著下降（分别为76.8%和53.6%对90.2%和77.2%;P〈0.05）。在冷应激条件下,猪卵母细胞CIRP mRNA表达水平升高,并在一定范围内随冷应激处理时间的延长而增加,至冷应激处理4 h时达最高。猪卵母细胞在短时间冷应激条件下,其发育能力和亚细胞结构并未受到显著影响;适当的冷应激条件可有效激发猪卵母细胞CIRP mRNA的表达。

英文摘要：

The purpose of this study is to investigate the effects of cold stress on the development in vitro and CIRP（ cold inducible RNA binding protein） gene expressions in pig oocytes. Porcine GV oocytes routinely cultured in vitro were exposure to room temperature（ 25 ℃）for cold stress treatment of 0. 5,1,2,6,12 and 24 h,respectively. Subsequent maturation of oocytes,embryo development and cytoskeletal structure were observed. CIRP mRNA expressions were analyzed by real-time PCR. The results showed that no significant differences were obsereved in the maturation rate,cleavage rate,microfilament and microtubule configuration when the oocytes were preserved for a short time（ 2 h）（ P〈0. 05）; however,the blastocyst development rate for cold stress treatment of 2 h significantly decreased as compared to the control（ 9. 7% vs 12. 3%,P〈0. 05）. Cold stress treatment for more than 2 h could result in evidently decreases in the maturation rate of oocytes and blastocyst development rate and the normal rate of subcellular configuration（ 76. 8% and 53. 6% versus 90. 2% and 77. 2%） as compared to the control（ P〈0. 05）. CIRP mRNA expressions of oocytes under cold stress were significantly increased with the treatment time and reached the top level for 4 h treatment. In vitro developmental competence and subcellular structure of porcine oocytes could not be affected by a short term cold stress and CIRP expression of porcine oocytes could be stimulated by cold stress.