A simplified one-step nuclear transfer procedure alters the gene expression patterns and developmental potential of cloned porcine embryos.
- Author:
Sang Kyu PARK
1
;
Sangho ROH
;
Jong Im PARK
Author Information
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords: CDX2; EOMES; HSP70; porcine; somatic cell nuclear transfer
- MeSH: Animals; Biological Markers/metabolism; Cloning, Organism; Embryo, Mammalian/metabolism; Female; *Gene Expression Regulation, Developmental; Nuclear Transfer Techniques/instrumentation/*veterinary; Oocytes/metabolism; Pregnancy; Real-Time Polymerase Chain Reaction; Swine/*embryology/*genetics
- From:Journal of Veterinary Science 2014;15(1):73-80
- CountryRepublic of Korea
- Language:English
- Abstract: Various somatic cell nuclear transfer (SCNT) techniques for mammalian species have been developed to adjust species-specific procedures to oocyte-associated differences among species. Species-specific SCNT protocols may result in different expression levels of developmentally important genes that may affect embryonic development and pregnancy. In the present study, porcine oocytes were treated with demecolcine that facilitated enucleation with protruding genetic material. Enucleation and donor cell injection were performed either simultaneously with a single pipette (simplified one-step SCNT; SONT) or separately with different pipettes (conventional two-step SCNT; CTNT) as the control procedure. After blastocysts from both groups were cultured in vitro, the expression levels of developmentally important genes (OCT4, NANOG, EOMES, CDX2, GLUT-1, PolyA, and HSP70) were analyzed by real-time quantitative polymerase chain reaction. Both the developmental rate according to blastocyst stage as well as the expression levels CDX2, EOMES, and HSP70 were elevated with SONT compared to CTNT. The genes with elevated expression are known to influence trophectoderm formation and heat stress-induced arrest. These results showed that our SONT technique improved the development of SCNT porcine embryos, and increased the expression of genes that are important for placental formation and stress-induced arrest.
