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钙在牛卵母细胞成熟和重编程中的作用:低钙模型的系统研究

The Role of Ca in Maturation and Reprogramming of Bovine Oocytes: A System Study of Low-Calcium Model.

作者信息

Meng Lin, Hu Hongmei, Liu Zhiqiang, Zhang Luyao, Zhuan Qingrui, Li Xue, Fu Xiangwei, Zhu Shien, Hou Yunpeng

机构信息

State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, China.

Hubei Key Laboratory of Natural Medicinal Chemistry and Resource Evaluation, School of Pharmacy, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.

出版信息

Front Cell Dev Biol. 2021 Oct 26;9:746237. doi: 10.3389/fcell.2021.746237. eCollection 2021.

Abstract

[Ca] is essential for mammalian oocyte maturation and early embryonic development, as those processes are Ca dependent. In the present study, we investigated the effect of [Ca] on maturation and reprogramming of oocytes in a lower calcium model of oocyte at metaphase II (MII) stage, which was established by adding cell-permeant Ca chelator BAPTA-AM to the maturation medium. Results showed that the extrusion of the first polar body (PB1) was delayed, and oocyte cytoplasmic maturation, including mitochondrial and endoplasmic reticulum distribution, was impaired in lower calcium model. The low-calcium-model oocytes presented a poor developmental phenotype of somatic cell nuclear transfer (SCNT) embryos at the beginning of activation of zygotic genome. At the same time, oxidative stress and apoptosis were observed in the low-calcium-model oocytes; subsequently, an RNA-seq analysis of the lower-calcium-model oocytes screened 24 genes responsible for the poor oocyte reprogramming, and six genes (ID1, SOX2, DPPA3, ASF1A, MSL3, and KDM6B) were identified by quantitative PCR. Analyzing the expression of these genes is helpful to elucidate the mechanisms of [Ca] regulating oocyte reprogramming. The most significant difference gene in this enriched item was ID1. Our results showed that the low calcium might give rise to oxidative stress and apoptosis, resulting in impaired maturation of bovine oocytes and possibly affecting subsequent reprogramming ability through the reduction of ID1.

摘要

钙离子([Ca])对于哺乳动物卵母细胞成熟和早期胚胎发育至关重要,因为这些过程依赖于钙离子。在本研究中,我们在卵母细胞处于中期II(MII)阶段的低钙模型中,研究了[Ca]对卵母细胞成熟和重编程的影响,该模型通过向成熟培养基中添加细胞渗透性钙螯合剂BAPTA-AM来建立。结果表明,在低钙模型中,第一极体(PB1)的排出延迟,并且卵母细胞的细胞质成熟,包括线粒体和内质网分布,均受到损害。在合子基因组激活开始时,低钙模型卵母细胞呈现出体细胞细胞核移植(SCNT)胚胎发育不良的表型。同时,在低钙模型卵母细胞中观察到氧化应激和细胞凋亡;随后,对低钙模型卵母细胞进行RNA测序分析,筛选出24个导致卵母细胞重编程不良的基因,并通过定量PCR鉴定出6个基因(ID1、SOX2、DPPA3、ASF1A、MSL3和KDM6B)。分析这些基因的表达有助于阐明[Ca]调节卵母细胞重编程的机制。该富集项中差异最显著的基因是ID1。我们的结果表明,低钙可能导致氧化应激和细胞凋亡,从而导致牛卵母细胞成熟受损,并可能通过降低ID1影响随后的重编程能力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b48e/8577575/844eaab4f5be/fcell-09-746237-g001.jpg

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