The aim of this study is to investigate whether endoplasmic reticulum (ER) stress attenuation could improve porcine somatic cell nuclear transfer (SCNT) embryo developmental competence. We treated porcine SCNT embryos with TUDCA (tauroursodeoxycholic acid, an inhibitor of ER stress) and/or TM (tunicamycin, an ER stress inducer), and examined embryonic developmental potential, embryo quality, the levels of ER stress markers (XBP1 protein and mRNA) and apoptosis-related-genes (BAX and BCL2 mRNA). Immunostaining detected X-box-binding protein (XBP1), a key gene regulator during ER stress, at all stages of SCNT embryo development. Embryo development analysis revealed that TUDCA treatment markedly increased (p<0.05) blastocyst formation rate, total cell number and inner cell mass (ICM) cell number compared to untreated control group. The TUDCA and TM groups showed significant alterations in XBP1 protein and XBP1-s mRNA levels compared to controls (lower and higher, respectively; p<0.05). Also, TUDCA treatment reduced oxidative stress by up-regulation of the antioxidant, GSH. TUNEL assay showed that TUDCA treatment significantly reduced apoptosis in porcine SCNT blastocysts confirmed by decreased pro-apoptotic BAX and increased anti-apoptotic BCL2 mRNA levels. Collectively, our results indicated that TUDCA can enhance the developmental potential of porcine SCNT embryos by attenuating ER-stress and reducing apoptosis.
Keywords: Apoptosis; Endoplasmic reticulum stress; Porcine SCNT embryos; TUDCA; XBP1.
Copyright © 2016 Society for Biology of Reproduction & the Institute of Animal Reproduction and Food Research of Polish Academy of Sciences in Olsztyn. Published by Elsevier Urban & Partner Sp. z o.o. All rights reserved.