Expression analysis of an α‐1, 3‐galactosyltransferase, an enzyme that creates xenotransplantation‐related α–Gal epitope, in pig preimplantation embryos
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 B4 (BS‐I‐B4). Staining with BS‐I‐B4 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.
Document Type: Research Article
Affiliations: 1: Laboratory of Animal Reproduction, Faculty of Agriculture 2: Section of Gene Expression Regulation, Frontier Science Research Center, Kagoshima University, Kagoshima, Japan
Publication date: January 1, 2012