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体外培养牛胚胎中的胚胎转录

Embryonic transcription in in vitro cultured bovine embryos.

作者信息

Barnes F L, First N L

机构信息

GenMark, Inc., Salt Lake City, Utah 84108.

出版信息

Mol Reprod Dev. 1991 Jun;29(2):117-23. doi: 10.1002/mrd.1080290205.

Abstract

The rate of cleavage and the onset of embryonic transcription of bovine embryos cultured in vitro (IVC) has been investigated. Embryos were derived from in vitro matured, in vitro fertilized oocytes (IVM/IVF) to improve developmental synchrony. The rate of cleavage was assessed by morphological evaluation between the one- and eight- to 16-cell stage. The rate of cleavage was found to be equivalent to that reported for in vivo recovered embryos. To assess the onset of embryonic transcription, embryos were cultured to the eight- to 16-cell stage in the presence of alpha-amanitin for various periods of time followed by two-dimensional polyacrylamide gel electrophoresis. Embryos readily cleaved to the eight- to 16-cell stage in the presence of inhibitor. alpha-Amanitin-sensitive protein synthesis was first detected at 36-48 h post-insemination (hpi) and continued up to 84 hpi. We conclude that bovine embryos produced by IVM/IVF/IVC are competent to initiate embryonic transcription at 36-48 h post-insemination and suggest that in vitro-induced cleavage arrest is not due to failure of the embryonic genome to initiate transcription.

摘要

对体外培养(IVC)的牛胚胎的卵裂率和胚胎转录起始情况进行了研究。胚胎来源于体外成熟、体外受精的卵母细胞(IVM/IVF),以提高发育同步性。通过在一细胞期和八至十六细胞期之间进行形态学评估来确定卵裂率。发现卵裂率与体内回收胚胎的报道相当。为了评估胚胎转录的起始情况,将胚胎在α-鹅膏蕈碱存在的情况下培养至八至十六细胞期不同时间,然后进行二维聚丙烯酰胺凝胶电泳。在抑制剂存在的情况下,胚胎很容易分裂至八至十六细胞期。在授精后36 - 48小时(hpi)首次检测到对α-鹅膏蕈碱敏感的蛋白质合成,并持续至84 hpi。我们得出结论,由IVM/IVF/IVC产生的牛胚胎在授精后36 - 48小时有能力启动胚胎转录,并表明体外诱导的卵裂停滞并非由于胚胎基因组启动转录失败所致。

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