title: Gene expression analysis in the compartments of the murine uterus.
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.
authors.affiliation: Developmental and Regenerative Biology, Institute of Medical Biology, A(⁎)STAR, 8A Biomedical Grove, #06-06 Immunos, Singapore 138648, Singapore. Electronic address: gracy.rosario@gmail.com.
authors.forename: Gracy Xavier
authors.initials: GX
authors.lastname: Rosario
authors.affiliation: UNC Neuroscience Center, 115 Mason Farm Road Campus, Box 7250, Chapel Hill, NC 27599-7250, USA.
authors.forename: Jr-Gang
authors.initials: JG
authors.lastname: Cheng
authors.affiliation: Developmental and Regenerative Biology, Institute of Medical Biology, A(⁎)STAR, 8A Biomedical Grove, #06-06 Immunos, Singapore 138648, Singapore.
authors.forename: Colin L
authors.initials: CL
authors.lastname: Stewart
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000378:metabolism; D010064:Embryo Implantation / Q000235:genetics*; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000235:genetics*; D051379:Mice; D011247:Pregnancy; D014176:Protein Biosynthesis / Q000235:genetics*; D012333:RNA, Messenger / Q000096:biosynthesis; D015398:Signal Transduction / Q000235:genetics; D014599:Uterus / Q000254:growth & development* / Q000378:metabolism
chemical_list: D012333:RNA, Messenger
keywords: Immunofluorescence; LIF; Luminal epithelium; Microarray; Stroma; Uterus