title: Expression analysis of an α-1, 3-galactosyltransferase, an enzyme that creates xenotransplantation-related α-Gal epitope, in pig preimplantation embryos.
abstract: α-1,3-Galactosyltransferase (α-GalT), an enzyme creating Galα1-3Gal (α-Gal) epitope on the cell surface in some mammalian species such as pigs, is known to be a key factor that causes hyperacute rejection upon transplantation from pigs to humans. To establish the RNA interference-based suppression of endogenous α-GalT messenger RNA (mRNA) synthesis in porcine preimplantation embryos, we determined the suitable embryonic stage at which stage such approach is possible by using the semi-quantitative RT-PCR (qRT-PCR) and the cytochemical method using a fluorescence-labeled Bandeiraea simplicifolia Isolectin B(4)  (BS-I-B(4) ). Staining with BS-I-B(4)  demonstrated that α-Gal epitope expression was first recognized at the 8-cell stage, and increased up to the hatched blastocyst stage. Single embryo-based qRT-PCR also confirmed this pattern. These results indicate that creation of α-Gal epitope is proceeded by de novo synthesis of α-GalT mRNA in porcine preimplantation embryos with peaking at the blastocyst stage.
authors.affiliation: Laboratory of Animal Reproduction, Faculty of Agriculture Section of Gene Expression Regulation, Frontier Science Research Center, Kagoshima University, Kagoshima, Japan. chy75jp@yahoo.co.jp
authors.forename: Haiying
authors.initials: H
authors.lastname: Chi
authors.forename: Masahiro
authors.initials: M
authors.lastname: Sato
authors.forename: Mitsutoshi
authors.initials: M
authors.lastname: Yoshida
authors.forename: Kazuchika
authors.initials: K
authors.lastname: Miyoshi
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000201:enzymology* / Q000378:metabolism*; D002478:Cells, Cultured; D004187:Disaccharides / Q000378:metabolism*; D000939:Epitopes; D005260:Female; D005307:Fertilization in Vitro; D005700:Galactosyltransferases / Q000235:genetics / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D006651:Histocytochemistry / Q000379:methods; D006801:Humans; D066298:In Vitro Techniques; D009865:Oocytes; D016133:Polymerase Chain Reaction / Q000379:methods; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D013552:Swine
chemical_list: D004187:Disaccharides; D000939:Epitopes; D012333:RNA, Messenger; C055075:galactosyl-(1-3)galactose; D005700:Galactosyltransferases; C530267:alpha-1,3-galactosyltransferase 1, porcine