earticle

영어

We examined the effect of endoplasmic reticulum (ER) stress inhibitor treatment during somatic cell nuclear transfer (SCNT) process on the reprogramming efficiency of porcine embryos. ER stress inhibitors such as Salubrinal (200 nM) and tauroursodeoxychloic acid (TUDCA, 100 μM) were added to the manipulation medium and holding medium. Porcine SCNT embryos were sampled at the 1-cell stage for mRNA extraction. The expression of x-box binding protein 1 (Xbp1), ER stress-associated genes and apoptotic genes were confirmed by RT-PCR or RT-qPCR. The levels of X-box binding protein 1 (Xbp1) splicing and the expression of ER stress-associated genes were significantly decreased by TUDCA treatment at the 1-cell stage of SCNT embryos (p<0.05). The expression of ER stress-associated genes was also slightly decreased by a combination treatment of salubrinal and TUDCA (Sal+TUD). The expression levels of Caspase-3 and Bcl2-associated X protein (Bax) mRNA were also significantly lower in TUDCA and Sal+TUD groups (p<0.05). The blastocyst formation rate (20.2%, 41/203) and the mean cells number in blastocysts (63.0±7.2, p<0.05) were significantly increased in the TUDCA treatment group compared with the control group (12.6% and 41.7±3.1, p<0.05). These results show that the treatment of ER stress inhibitor, especially TUDCA, during SCNT process can inhibit cellular damages and enhance in vitro development of SCNT embryos by reducing ER stress.