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寻找小鼠ped基因的牛同源物及其在牛植入前发育过程中信使核糖核酸表达的特征分析。

Search for the bovine homolog of the murine ped gene and characterization of its messenger RNA expression during bovine preimplantation development.

作者信息

Fair Trudee, Gutierrez-Adan Alfonso, Murphy Madeline, Rizos Dimitrios, Martin Finian, Boland Maurice P, Lonergan Patrick

机构信息

Department of Animal Science and Production, University College Dublin, Lyons Research Farm, Newcastle, County Dublin, Ireland.

出版信息

Biol Reprod. 2004 Feb;70(2):488-94. doi: 10.1095/biolreprod.103.022137. Epub 2003 Oct 20.

Abstract

In mice, a gene (Ped: preimplantation embryo development) that regulates preimplantation embryonic growth, including cleavage rate and embryo survivability, has been described. The objective of the current study was to identify the bovine homolog of the Ped gene and to characterize the mRNA expression pattern of this gene during bovine preimplantation embryo development. The NCBI GenBank/EBI expressed sequence tags (EST) databases were searched for bovine ESTs that were homologous to the murine Ped gene, and the resulting ESTs were aligned and assembled into a contiguous sequence. The homology of the sequence to the murine Ped gene was confirmed. Primers were designed for the sequence, and the mRNA expression pattern was characterized during bovine preimplantation embryo development in vivo and in vitro. In vitro-produced bovine zygotes were cultured either in vitro, in synthetic oviduct fluid, or in vivo in the ewe oviduct for 1-7 days and processed for quantitative real-time polymerase chain reaction (PCR). Transcript abundance increased at each stage of development. However, the expression levels were consistently higher in in vivo-cultured embryos at all stages, with in vivo-cultured embryos showing a 9-fold increase in relative transcript abundance during culture from the zygote to the blastocyst stage in contrast to just under a 4-fold increase during the same culture period in vitro. The mRNA expression pattern of the gene was investigated in early- and late-cleaving two-cell embryos collected at 25, 28, 32, and >or=36 h postinsemination (pi). Transcript relative abundance was highest in those embryos that had cleaved by 28 hpi and decreased almost 3-fold thereafter. In conclusion, we have identified a potential bovine homolog of the murine Ped gene. We have characterized the mRNA expression pattern of this gene during preimplantation embryo development in cattle and shown that a greater relative abundance of the gene transcript is associated with embryos of higher quality (in vivo cultured) and greater developmental potential (early cleaving).

摘要

在小鼠中,已发现一个调控着床前胚胎生长(包括卵裂率和胚胎存活率)的基因(Ped:着床前胚胎发育基因)。本研究的目的是鉴定牛的Ped基因同源物,并描述该基因在牛着床前胚胎发育过程中的mRNA表达模式。在NCBI GenBank/EBI表达序列标签(EST)数据库中搜索与小鼠Ped基因同源的牛EST,将得到的EST进行比对并组装成一个连续序列。确认了该序列与小鼠Ped基因的同源性。为该序列设计引物,并在体内和体外牛着床前胚胎发育过程中对mRNA表达模式进行了表征。体外生产的牛受精卵在体外、合成输卵管液中或在母羊输卵管内体内培养1至7天,然后进行定量实时聚合酶链反应(PCR)。转录本丰度在发育的每个阶段都增加。然而,在所有阶段,体内培养的胚胎中的表达水平始终较高,体内培养的胚胎从合子到囊胚阶段培养期间相对转录本丰度增加了9倍,而在相同培养期体外培养时仅增加了不到4倍。在授精后25、28、32和≥36小时收集的早期和晚期卵裂的二细胞胚胎中研究了该基因的mRNA表达模式。转录本相对丰度在28小时授精后卵裂的胚胎中最高,此后几乎下降了3倍。总之,我们鉴定出了小鼠Ped基因的一个潜在牛同源物。我们已经描述了该基因在牛着床前胚胎发育过程中的mRNA表达模式,并表明该基因转录本的相对丰度更高与更高质量(体内培养)和更大发育潜力(早期卵裂)的胚胎相关。

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