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卵母细胞供体年龄和胚胎发育阶段对牛体外生产胚胎中前列腺素和孕酮合成途径相关酶编码基因转录的影响。

Effects of oocyte donor age and embryonic stage of development on transcription of genes coding for enzymes of the prostaglandins and progesterone synthesis pathways in bovine in vitro produced embryos.

作者信息

Torres A, Batista M, Diniz P, Silva E, Mateus L, Lopes-da-Costa L

机构信息

Reproduction and Obstetrics,CIISA,Faculty of Veterinary Medicine,University of Lisbon,Avenida da Universidade Técnica,Alto da Ajuda,1300-477 Lisboa,Portugal.

Reproduction and Obstetrics,CIISA,Faculty of Veterinary Medicine,Universidade de Lisboa,Avenida da Universidade Técnica,Alto da Ajuda,1300-477 Lisboa,Portugal.

出版信息

Zygote. 2015 Dec;23(6):802-12. doi: 10.1017/S0967199414000446. Epub 2014 Sep 26.

Abstract

The ability of early bovine embryos to produce prostaglandins (PGs) and progesterone (P4), and the role of these mediators in embryonic development and survival are poorly understood. In this study we tested the hypothesis that day 7 bovine embryos are able to transcribe genes coding for enzymes of the PGs (PTGS2, PGES, PGFS) and P4 (StAR, P450scc, 3β-HSD) synthesis pathways, and that transcription levels of these genes are associated with developmental progression and heifer age-related [pre-pubertal (PP) versus post-pubertal cyclic (C)] oocyte competence. Compared with C heifer oocytes, PP heifer oocytes showed a lower (P < 0.0001) in vitro blastocyst rate, but in embryos developing until day 7, heifer age had no effect on quality grade. Day 7 quality grade 1-2 embryos were selected for RNA extraction and gene transcription analysis by qRT-PCR, in a 2 × 2 factorial design [age (PP or C) × embryonic stage (compact morulae and early blastocysts, CM + EBL, or blastocysts and expanded blastocysts, BL + BEX); 15 embryos/group]. Transcription levels of PTGS2, PGES, PGFS, P450scc and 3β-HSD were not affected by heifer age but were higher (P < 0.01) in BL + BEX than in CM + EBL. In conclusion, the main limiting factor for embryo production from PP heifers is oocyte competence. Day 7 bovine embryos evidence transcription of genes coding for enzymes of PGs and P4 synthesis pathways, and transcription levels are associated with blastocyst differentiation. This prompts for an autocrine/paracrine action of PGs and P4 in early bovine embryonic development.

摘要

早期牛胚胎产生前列腺素(PGs)和孕酮(P4)的能力,以及这些介质在胚胎发育和存活中的作用,目前了解甚少。在本研究中,我们检验了以下假设:第7天的牛胚胎能够转录编码PGs(PTGS2、PGES、PGFS)和P4(StAR、P450scc、3β-HSD)合成途径中酶的基因,并且这些基因的转录水平与发育进程以及与小母牛年龄相关的[青春期前(PP)与青春期后周期性(C)]卵母细胞能力有关。与C组小母牛卵母细胞相比,PP组小母牛卵母细胞的体外囊胚率较低(P < 0.0001),但在发育至第7天的胚胎中,小母牛年龄对质量等级没有影响。选择第7天质量等级为1-2的胚胎进行RNA提取,并通过qRT-PCR进行基因转录分析,采用2×2析因设计[年龄(PP或C)×胚胎阶段(致密桑葚胚和早期囊胚,CM + EBL,或囊胚和扩张囊胚,BL + BEX);每组15个胚胎]。PTGS2、PGES、PGFS、P450scc和3β-HSD的转录水平不受小母牛年龄的影响,但在BL + BEX组中高于CM + EBL组(P < 0.01)。总之,PP小母牛胚胎生产的主要限制因素是卵母细胞能力。第7天的牛胚胎证明了编码PGs和P4合成途径中酶的基因的转录,并且转录水平与囊胚分化有关。这提示了PGs和P4在早期牛胚胎发育中的自分泌/旁分泌作用。

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