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热应激对牛卵母细胞周围卵丘细胞在成熟过程中的影响:基质金属蛋白酶 9 和孕酮产生的改变。

Heat stress effects on the cumulus cells surrounding the bovine oocyte during maturation: altered matrix metallopeptidase 9 and progesterone production.

机构信息

Department of Animal Science, Institute of Agriculture, University of Tennessee, UT AgResearch, 251 Brehm Animal Science Building, 2506 River Drive, Knoxville, Tennessee 37996-4574, USA.

出版信息

Reproduction. 2013 Aug 1;146(2):193-207. doi: 10.1530/REP-12-0487. Print 2013 Aug.

Abstract

When the effects of heat stress are detrimental during maturation, cumulus cells are intimately associated with the oocyte. To determine the extent to which heat stress affects these cells, in this study, transcriptome profiles of the cumulus that surrounded control and heat-stressed oocytes (41 °C during the first 12 h only and then shifted back to 38.5 °C) during in vitro maturation (IVM) were compared using Affymetrix bovine microarrays. The comparison of cumulus-derived profiles revealed a number of transcripts whose levels were increased (n=11) or decreased (n=13) ≥ twofold after heat stress exposure (P<0.01), sufficient to reduce the development of blastocysts by 46.4%. In a separate study, quantitative PCR (qPCR) was used to confirm heat-induced differences in the relative abundances of the transcripts of five different genes (caveolin 1, matrix metallopeptidase 9, FSH receptor, Indian hedgehog homolog, and inducible nitric oxide synthase). Heat stress exposure resulted in >1.7-fold decrease in the protein levels of latent matrix metallopeptidase 9 (proMMP9). Heat-induced reductions in transcript levels were noted at 6 h IVM with reductions in proMMP9 protein levels at 18 h IVM (P=0.0002). Independent of temperature, proMMP9 levels at 24 h IVM were positively correlated with the development rate of blastocysts (R²=0.36; P=0.002). The production of progesterone increased during maturation; heat-induced increases were evident by 12 h IVM (P=0.002). Both MMP9 and progesterone are associated with the developmental competence of the oocyte; thus, it seems plausible for some of the negative consequences of heat stress on the cumulus-oocyte complex to be mediated through heat-induced perturbations occurring in the surrounding cumulus.

摘要

当热应激在成熟过程中产生不利影响时,卵丘细胞与卵母细胞密切相关。为了确定热应激对这些细胞的影响程度,本研究使用 Affymetrix 牛微阵列比较了体外成熟(IVM)期间受控制和受热应激(仅在最初 12 小时内 41°C,然后恢复到 38.5°C)的卵母细胞周围卵丘的转录组谱。对卵丘衍生谱的比较揭示了许多转录物的水平增加(n=11)或减少(n=13)≥两倍后热应激暴露(P<0.01),足以使囊胚的发育减少 46.4%。在另一项研究中,使用定量 PCR(qPCR)来确认五种不同基因(窖蛋白 1、基质金属蛋白酶 9、FSH 受体、印度刺猬同源物和诱导型一氧化氮合酶)的转录物相对丰度的热诱导差异。热应激暴露导致潜伏基质金属蛋白酶 9(proMMP9)的蛋白水平降低 1.7 倍以上。在 IVM 6 小时时观察到热诱导的转录水平降低,并且在 IVM 18 小时时 proMMP9 蛋白水平降低(P=0.0002)。与温度无关,IVM 24 小时时 proMMP9 水平与囊胚发育率呈正相关(R²=0.36;P=0.002)。孕酮的产生在成熟过程中增加;在 IVM 12 小时时可以明显观察到热诱导的增加(P=0.002)。MMP9 和孕酮都与卵母细胞的发育能力有关;因此,热应激对卵丘-卵母细胞复合体的一些负面影响似乎可以通过周围卵丘中发生的热诱导扰动来介导。

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