<em>TNF</em>-<em>α</em>在牦牛卵母细胞及早期体外受精胚胎中的表达

doi:10.3969/j.issn.1674-7968.2019.08.010

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Abstract Tumor necrosis factor α (TNF-α) is a pro-inflammatory Th1 cytokine that is involved in the regulation of mammalian ovulation, embryonic development and implantation. The aim of this study was to investigate the role of TNF-α in the development of oocyte maturation and in vitro fertilization (IVF) embryos by detecting the expression of TNF-α in yak (Bos grunneins) oocyte maturation and early IVF embryo development. In this study, the cumulus-oocyte complex (COCs) were collected and matured in vitro and fertilized to obtain IVF embryos, and to evaluate the developmental ability of embryos at different stages of implantation. Methods of qRT-PCR, indirect immunofluorescence staining and Western blot were used to detect the expression and distribution of TNF-α in immature oocytes, mature oocytes and embryos at different stages. The results showed as follows: (1) The rates of embryos developed to stages of 2~4 cells, 5~8 cells, morulae were (61.59±0.31)%, (53.76±0.22)% and (26.88±0.32)%, respectively, and the blastocyst rate was (16.12±0.23)%. (2) qRT-PCR results showed that, TNF-α mRNA could be detected in immature COCs, mature COCs, morulae and blastocysts, the relative expression levels in the order of high to low was mature COCs, immature COCs, blastocysts and morulae. 2~4 cells and 5~8 cells embryos had no expression. (3) The result of indirect immunofluorescence and Western blot showed that the protein of TNF-α was mainly distributed in the cytoplasm of COCs and embryos, which was weaker in the nucleus. The protein of TNF-α could be detected in COCs, morula and blastocyst, the relative expression levels in the order of high to low was mature COCs, immature COCs, blastocysts and morulae, but 2~4 cells and 5~8 cells embryos had no expression. The results in this study indicated that TNF-α might play a role in the process of the maturation and senescence of the yak oocytes and might be involved in the process of blastocyst cell apoptosis and embryo implantation, which provides a theoretical basis for revealing the mechanism of TNF-α in yak reproduction.

Key words： Tumor necrosis factor alpha (TNF-α) Yak (Bos grunneins) In vitro Fertilization (IVF) Embryo development

Received: 24 January 2019

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Corresponding Authors: * , sjyu@163.com

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	XU Tao
	PAN Yang-Yang
	HE Hong-Hong
	LI Gu-Yue
	CHEN Ping
	WEN Ze-Xing
	CUI Yan
	YU Si-Jiu

Cite this article:

XU Tao,PAN Yang-Yang,HE Hong-Hong, et al. The Expression of TNF-α in Yak (Bos grunniens) Oocytes and Preimplantation Embryos Produced by In vitro Fertilization[J]. 农业生物技术学报, 2019, 27(8): 1423-1433.

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http://journal05.magtech.org.cn/Jwk_ny/EN/10.3969/j.issn.1674-7968.2019.08.010 OR http://journal05.magtech.org.cn/Jwk_ny/EN/Y2019/V27/I8/1423

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