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小鼠子宫各部分的基因表达分析。

Gene expression analysis in the compartments of the murine uterus.

作者信息

Rosario Gracy Xavier, Cheng Jr-Gang, Stewart Colin L

机构信息

Developmental and Regenerative Biology, Institute of Medical Biology, A(⁎)STAR, 8A Biomedical Grove, #06-06 Immunos, Singapore 138648, Singapore.

UNC Neuroscience Center, 115 Mason Farm Road Campus, Box 7250, Chapel Hill, NC 27599-7250, USA.

出版信息

Differentiation. 2016 Apr-Jun;91(4-5):42-9. doi: 10.1016/j.diff.2015.10.009. Epub 2015 Dec 2.

Abstract

Embryo implantation, a key critical feature of mammalian pregnancy, involves co-ordinate interplay between an incoming blastocyst and a receptive uterus. Aberrations in signaling cascades during this process result in pregnancy loss in mammals, including women. Analysis of the complete uterus at any given point either during preparation for implantation or during and after embryo attachment and invasion makes it difficult to assign specific signaling mechanism to the individual cellular compartments of the uterus. Here, we describe methods for the specific isolation of the luminal epithelium (LE) and subsequent analysis of gene expression/signaling pathways during embryo attachment. We further describe the analysis of RNA and proteins by specific techniques of quantitative PCR (qPCR), immunostaining and Western blotting of uterine tissues. These methods can be applied to the other cellular compartments of the uterus and embryo invasion and endometrial development. These techniques will be beneficial to investigators for delineating the mechanisms involved during embryo attachment and female reproduction as well as providing a means to studying highly dynamic changes in gene expression in tissues.

摘要

胚胎着床是哺乳动物妊娠的一个关键重要特征,涉及植入的囊胚与处于接受态的子宫之间的协调相互作用。在此过程中信号级联的异常会导致包括女性在内的哺乳动物妊娠失败。在植入准备期间或胚胎附着及侵入期间和之后的任何给定时间对整个子宫进行分析,使得难以将特定的信号传导机制归因于子宫的各个细胞区室。在这里,我们描述了特异性分离腔上皮(LE)以及随后在胚胎附着期间分析基因表达/信号通路的方法。我们还通过定量PCR(qPCR)、免疫染色和子宫组织的蛋白质印迹等特定技术描述了对RNA和蛋白质的分析。这些方法可应用于子宫的其他细胞区室以及胚胎侵入和子宫内膜发育。这些技术将有助于研究人员阐明胚胎附着和雌性生殖过程中涉及的机制,并提供一种研究组织中基因表达高度动态变化的手段。

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