Department of Obstetrics and Reproduction, Veterinary Faculty, University of Thessaly, Karditsa, Greece.
Department of Obstetrics and Reproduction, Veterinary Faculty, University of Thessaly, Karditsa, Greece; Department of Genetics, Comparative and Evolutionary Biology, Faculty of Biochemistry and Biotechnology, University of Thessaly, Larissa, Greece.
Theriogenology. 2020 Oct 15;156:36-45. doi: 10.1016/j.theriogenology.2020.06.039. Epub 2020 Jun 30.
Heat stress causes subfertility in cattle by inducing alterations in steroidogenic capacity, follicular function and ovulation defects, which eventually negatively affect oocyte quality and embryo survival. Here, the effects of short, moderate temperature elevation during IVM, on embryo yield, and on the expression of various genes was evaluated. In 8 replicates, cumulus oocyte complexes (COCs) were matured for 24 h at 39 °C (controls n = 605) or at 41 °C from hour 2 to hour 8 of IVM (treated, n = 912), fertilized, and presumptive zygotes were cultured for 9 days at 39 °C. Cleavage and embryo formation rates were evaluated 48 h post insemination and on days 7, 8, 9 respectively. Cumulus cells, oocytes and blastocysts from 5 replicates were snap frozen for the relative expression analysis of genes related to metabolism, thermal and oxidative stress response, apoptosis, and placentation. In treated group, cleavage and embryo formation rates were statistically significantly lower compared with the control (cleavage 86.7% vs 74.2%; blastocysts: day 7, 29.9% vs 19.7%, day 8, 34.2% vs 22.9% and day 9 35.9% vs 24.5%). Relative mRNA abundance of three genes in cumulus cells (HSP90AA1, CPT1B, G6PD) and three genes in blastocysts (DNMT3A, PLAC8, GPX1) indicated significantly different expression between groups (p < 0.05)., The expression of G6PD, SOD2, GXP1 in oocytes and PTGS2 in blastocysts tended to differ among groups (0.05<p < 0.08). Heat stress altered (p < 0.05) the correlation of expression between HSPs and other genes in oocytes (G6PD, GPX1, CCNB1), cumulus cells (LDH, CCNB1) and blastocysts (AKR1B1, PLAC8). These results imply that exposure of oocytes to elevated temperature, even for only 6 h, disrupts the developmental competence of the oocytes, suppresses blastocyst yield and significantly alters the coordinated pattern of gene expressions.
热应激通过诱导甾体生成能力、卵泡功能和排卵缺陷的改变,导致牛的繁殖力下降,最终对卵母细胞质量和胚胎存活产生负面影响。在这里,评估了 IVM 期间短时间、中度温度升高对胚胎产量以及各种基因表达的影响。在 8 个重复中,卵丘-卵母细胞复合物(COC)在 39°C(对照,n=605)或 IVM 第 2 小时至第 8 小时的 41°C 下成熟 24 小时(处理,n=912),受精,假定的受精卵在 39°C 下培养 9 天。在受精后 48 小时评估卵裂和胚胎形成率,并分别在第 7、8 和 9 天进行评估。来自 5 个重复的卵丘细胞、卵母细胞和囊胚被立即冷冻,用于与代谢、热应激和氧化应激反应、细胞凋亡和胎盘形成相关的基因的相对表达分析。在处理组中,与对照组相比,卵裂和胚胎形成率明显降低(卵裂率 86.7%对 74.2%;囊胚:第 7 天 29.9%对 19.7%,第 8 天 34.2%对 22.9%,第 9 天 35.9%对 24.5%)。卵丘细胞中 3 个基因(HSP90AA1、CPT1B、G6PD)和囊胚中 3 个基因(DNMT3A、PLAC8、GPX1)的相对 mRNA 丰度表明组间表达差异显著(p<0.05)。卵母细胞中的 G6PD、SOD2、GPX1 和囊胚中的 PTGS2 的表达在组间也存在差异(0.05<p<0.08)。热应激改变了(p<0.05)卵母细胞中 HSPs 与其他基因(G6PD、GPX1、CCNB1)、卵丘细胞(LDH、CCNB1)和囊胚(AKR1B1、PLAC8)之间表达的相关性。这些结果表明,即使仅将卵母细胞暴露于高温 6 小时,也会破坏卵母细胞的发育能力,抑制囊胚产量,并显著改变基因表达的协调模式。