Developmental Characteristics of SCNT Pig Embryos Knocked-out of Alpha-1,3-Galactosyltransferase Gene

서지정보

ㆍ발행기관 : 한국동물번식학회 ㆍ수록지정보 : Reproductive & developmental biology / 33권 / 3호
ㆍ저자명 : Joo-Hyun Shim1, Mi-Rung Park, Byoung-Chul Yang, Yeoung-Gyu Ko, Keon Bong Oh, Jeong-Woong Lee, Jae-Seok Woo, Eung-Woo Park, Soo-Bong Park, Seongsoo Hwang

목차

ABSTRACT INTRODUCTION MATERIALS AND METHODS Chemicals In Vitro Maturation of Oocytes Nuclear Transfer Procedure Collection of In Vivo Blastocysts TUNEL Assay for Apoptotic Cell Death Apoptosis Related Gene Expression Statistical Analysis RESULTS In Vitro Maturation of Porcine Oocytes In Vitro Development of SCNT Embryos Apoptosis Rate in mNT and GalT KO NT Blastocysts Analysis of Apoptosis Gene Expression in Blastocysts DISCUSSION REFERENCES

영어 초록

This study was performed to comprehend the developmental characteristics of cloned embryos knocked out (KO) of α-1,3-galactosyltransferase (GalT) gene. Immature oocytes were collected and cultured for 40 hrs (1-step) or 20hrs (with hormone) + 20hrs (without hormone) (2-step). The embryos transferred with miniature pig ear fibroblast cell were used as control. The reconstructed embryos were cultured in PZM-3 with 5% CO2 in air at 38.5℃ for 6 days. To determine the quality of the blstocysts, TUNEL and quantitative realtime RT-PCR were performed. The embryos were transferred to a surrogate (Landrace) at an earlier stage of the estrus cycle. The maturation rate was significantly higher in 2-step method than that of 1-step (p<0.05). The blastocyst development of GalT KO embryos was significantly lower than that of normal cloned embryos (p<0.05). The total and apoptotic cell number of GalT KO blastocysts was not different statistically from control. The relative abundance of Bax-α/Bcl-xl ratio was significantly higher in both cloned blastocysts than that of in vivo blastocysts (p<0.05). Taken together, it can be postulated that the lower developmental potential and higher expression of apoptosis related genes in GalT KO SCNT embryos might be a cause of a low efficiency of GalT KO cloned miniature pig production.

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