학술논문
Expression of Bovine Ecat1 Gene in Immature and in vitro Matured Oocytes as Well as During Early Embryonic Development.
Document Type
Article
Source
Subject
*SLAUGHTERING
*GENE expression
*EMBRYOLOGY
*EMBRYONIC stem cells
*REVERSE transcriptase polymerase chain reaction
*BLASTOCYST
*IN vitro studies
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Language
ISSN
0936-6768
Abstract
Contents Ecat1 is a maternal effect gene that is exclusively expressed in oocytes and embryonic stem cells, and has an important role in pre-implantation development. This study was designed to investigate the expression of bovine Ecat1 gene in immature and in vitro matured oocytes as well as during early embryonic development, and also Ecat1 protein localization. Samples were obtained from slaughtered animals. RNA extractions were carried out from ovary, immature and in vitro matured oocytes and also different stages of embryonic development (2-, 4-, 8- to 16-cell stages and blastocysts). RT-PCR analysis revealed the expression of Ecat1 in ovary, oocytes and embryos. Analysis in FGENESH online tool predicted three exons and one transcription start site (TSS) in Ecat1 gene, and the 3′ RACE-PCR result showed that just one splice variant was amplified. By quantitative real-time PCR technique, we showed that Ecat1 transcript increased at 8- to 16-cell-stage embryos and decreased in blastocyst stage (p < 0.05). Immunofluorescence analysis showed cytoplasmic localization of Ecat1 protein in bovine oocytes. Results demonstrated bovine Ecat1 expression at protein level and also indicated that Ecat1 has a significant higher embryonic expression at 8- to 16-cell stage. This embryonic expression is probably required for further developmental stages. [ABSTRACT FROM AUTHOR]