Calder M D, Madan P, Watson A J
Department of Physiology and Pharmacology, University of Western Ontario, Children's Health Research Institute - LHRI, 5th Floor Victoria Research Laboratories, 800 Commissioners Road, London, Ontario, Canada N6A 4G5.
Zygote. 2008 May;16(2):161-8. doi: 10.1017/S096719940700456X.
RNA-binding proteins (RBP) influence RNA editing, localization, stability and translation and may contribute to oocyte developmental competence by regulating the stability and turnover of oogenetic mRNAs. The expression of Staufen 1 and 2 and ELAVL1, ELAVL2 RNA-binding proteins during cow early development was characterized. Cumulus-oocyte complexes were collected from slaughterhouse ovaries, matured, inseminated and subjected to embryo culture in vitro. Oocyte or preimplantation embryo pools were processed for RT-PCR and whole-mount immunofluorescence analysis of mRNA expression and protein distribution. STAU1 and STAU2 and ELAVL1 mRNAs and proteins were detected throughout cow preimplantation development from the germinal vesicle (GV) oocyte to the blastocyst stage. ELAVL2 mRNAs were detectable from the GV to the morula stage, whereas ELAVL2 protein was in all stages examined and localized to both cytoplasm and nuclei. The findings provide a foundation for investigating the role of RBPs during mammalian oocyte maturation and early embryogenesis.
RNA结合蛋白(RBP)影响RNA编辑、定位、稳定性和翻译,并可能通过调节卵母细胞mRNA的稳定性和周转来促进卵母细胞的发育能力。对牛早期发育过程中Staufen 1和2以及ELAVL1、ELAVL2 RNA结合蛋白的表达进行了表征。从屠宰场的卵巢中收集卵丘-卵母细胞复合体,使其成熟、受精并进行体外胚胎培养。对卵母细胞或植入前胚胎池进行处理,用于mRNA表达和蛋白质分布的RT-PCR及全胚胎免疫荧光分析。在牛从生发泡(GV)卵母细胞到囊胚阶段的植入前发育过程中,均检测到了STAU1、STAU2和ELAVL1的mRNA和蛋白质。ELAVL2的mRNA在从GV期到桑葚胚期均可检测到,而ELAVL2蛋白在所有检测阶段均有表达,并定位于细胞质和细胞核。这些发现为研究RBP在哺乳动物卵母细胞成熟和早期胚胎发生过程中的作用奠定了基础。