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Expression analysis of an α‐1, 3‐galactosyltransferase, an enzyme that creates xenotransplantation‐related α–Gal epitope, in pig preimplantation embryos
Author(s) -
CHI Haiying,
SATO Masahiro,
YOSHIDA Mitsutoshi,
MIYOSHI Kazuchika
Publication year - 2012
Publication title -
animal science journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.606
H-Index - 38
eISSN - 1740-0929
pISSN - 1344-3941
DOI - 10.1111/j.1740-0929.2011.00964.x
Subject(s) - xenotransplantation , epitope , blastocyst , embryo , microbiology and biotechnology , biology , messenger rna , transplantation , embryonic stem cell , andrology , embryogenesis , antigen , immunology , gene , biochemistry , medicine
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.