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促黄体生成素在排卵前激增,随后牛卵泡中 CYP19A1、HSD3B1 和 CYP17A1 的表达下调,相应启动子的染色质发生凝聚。

The pre-ovulatory luteinizing hormone surge is followed by down-regulation of CYP19A1, HSD3B1, and CYP17A1 and chromatin condensation of the corresponding promoters in bovine follicles.

机构信息

Leibniz Institute for Farm Animal Biology (FBN), Dummerstorf, Germany.

出版信息

Mol Reprod Dev. 2010 Dec;77(12):1040-8. doi: 10.1002/mrd.21257.

Abstract

The pre-ovulatory luteinizing hormone (LH) surge induces an extensive molecular, physiological, and morphological reorganization of the bovine follicle. This study was designed to elucidate if chromatin modulation is involved in the LH-induced gene regulation. Granulosa and theca of well-characterized large bovine follicles were isolated before and after the LH surge. CYP19A1, HSD3B1, and CYP17A1 transcripts, which encode key enzymes of steroid hormone biosynthesis, were quantified by real-time PCR (qPCR) and the degree of chromatin condensation was determined by DNase I protection assays. After LH, granulosa-specific CYP19A1 and theca-specific CYP17A1 transcripts were almost completely down-regulated. Also, the abundance of HSD3B1 transcripts was reduced. The promoter chromatin of HSD3B1 and particularly of CYP19A1 was significantly less accessible to DNAse I in both cell types after LH, whereas the chromatin accessibility of the CYP17A1 promoter changed only in the theca. Correlation analysis revealed partly, highly significant negative correlations between transcript abundance and protection from DNase I digestion of the corresponding chromatin. The data strongly suggest that LH induces cell type- and gene-specific chromatin condensation in the pre-ovulatory bovine follicle. This epigenetic mechanism might be involved in the pre-ovulatory down-regulation of genes.

摘要

促黄体激素(LH)排卵前激增会引起牛卵泡的广泛的分子、生理和形态重组。本研究旨在阐明染色质调节是否参与 LH 诱导的基因调控。在 LH 激增前后,从特征明确的大卵泡中分离出颗粒细胞和膜细胞。通过实时 PCR(qPCR)定量测定编码类固醇激素生物合成关键酶的 CYP19A1、HSD3B1 和 CYP17A1 转录本,并用 DNase I 保护试验测定染色质浓缩程度。LH 后,颗粒细胞特异性的 CYP19A1 和膜细胞特异性的 CYP17A1 转录本几乎完全下调。HSD3B1 转录本的丰度也降低了。LH 后,两种细胞类型中 HSD3B1 特别是 CYP19A1 的启动子染色质对 DNAse I 的可及性显著降低,而 CYP17A1 启动子的染色质可及性仅在膜细胞中发生变化。相关性分析显示,相应染色质的转录本丰度与 DNase I 消化保护之间存在部分显著负相关。这些数据强烈表明,LH 在牛卵泡排卵前诱导细胞类型和基因特异性染色质浓缩。这种表观遗传机制可能参与排卵前基因的下调。

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