Effect of Endoplasmic Reticulum (ER) Stress Inhibitor Treatment during Parthenogenetic Activation on the Apoptosis and In Vitro Development of Parthenogenetic Porcine Embryos
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We investigate the effect of endoplasmic reticulum (ER) stress inhibitor treatment during parthenogenetic ac-tivation of oocytes on the ER stress generation, apoptosis, and in vitro development of parthenogenetic porcine embryos. Por-cine in vitro matured oocytes were activated by 1) electric stimulus (E) or 2) E+10 μM Ca-ionophore (A23187) treatment (EC). Oocytes were then treated by ER stress inhibitors such as salubrinal (200 nM) and tauroursodeoxychloic acid (TUDCA, 100 μM) for 3 h prior to in vitro culture. Parthenogenetic embryos were sampled to analyze ER stress and apoptosis at the 1-cell and blastocyst stages. The x-box binding protein 1 (Xbp1) mRNA and ER stress-associated genes were analyzed by RT-PCR or RT-qPCR. Apoptotic gene expression was analyzed by RT-PCR. At the 1-cell stage, although no difference was ob-served in Xbp1 splicing among treatments, BiP transcription level in the E group was significantly reduced by salubrinal treatment, and GRP94 and ATF4 transcription levels in EC group were significantly reduced by all treatments (p<0.05) com-pared to control. In the EC group, both apoptotic genes were reduced by ER stress inhibitor treatments compared to control (p<0.05) except Caspase-3 gene by TUDCA treatment. These results suggest that the treatment of ER stress inhibitor during parthenogenetic activation can reduce ER stress, and thereby reduce apoptosis and promote in vitro development of porcine parthenogenetic embryos.
Abstract
We investigate the effect of endoplasmic reticulum (ER) stress inhibitor treatment during parthenogenetic ac-tivation of oocytes on the ER stress generation, apoptosis, and in vitro development of parthenogenetic porcine embryos. Por-cine in vitro matured oocytes were activated by 1) electric stimulus (E) or 2) E+10 μM Ca-ionophore (A23187) treatment (EC). Oocytes were then treated by ER stress inhibitors such as salubrinal (200 nM) and tauroursodeoxychloic acid (TUDCA, 100 μM) for 3 h prior to in vitro culture. Parthenogenetic embryos were sampled to analyze ER stress and apoptosis at the 1-cell and blastocyst stages. The x-box binding protein 1 (Xbp1) mRNA and ER stress-associated genes were analyzed by RT-PCR or RT-qPCR. Apoptotic gene expression was analyzed by RT-PCR. At the 1-cell stage, although no difference was ob-served in Xbp1 splicing among treatments, BiP transcription level in the E group was significantly reduced by salubrinal treatment, and GRP94 and ATF4 transcription levels in EC group were significantly reduced by all treatments (p<0.05) com-pared to control. In the EC group, both apoptotic genes were reduced by ER stress inhibitor treatments compared to control (p<0.05) except Caspase-3 gene by TUDCA treatment. These results suggest that the treatment of ER stress inhibitor during parthenogenetic activation can reduce ER stress, and thereby reduce apoptosis and promote in vitro development of porcine parthenogenetic embryos.
참고문헌 (18)
- Beuers U, "Effects of tauroursodeoxycholic acid on cytosolic Ca2+ signals in isolated rat hepatocytes", Gastroenterology, 104, pp.604~612 (1993)
- Beuers U, "Tauroursodeoxycholic acid stimulates hepatocellular exocytosis and mobilizes extracellular Ca2+ mechanisms defective in cholestasis", J Clin Invest, 92, pp.2984~2993 (1993)
- Boyce M, "A selective inhibitor of eIF2α dephosphorylationprotects cells from ER stress", Science, 307, pp.935~939 (2005)
- Choi JY, "Effect of activation time on the nuclear remodeling and in vitro development of nuclear transfer embryos derived from bovine somatic cells", Mol Reprod Dev, 69, pp.89~295 (2004)
- 황인선, "Mitochondrial and DNA damage in bovine somatic cell nuclear transfer embryos", Journal of Veterinary Science, 14(3), pp.235~240 (2013), 대한수의학회[DOI]
- 황인선, "Generation of Reactive Oxygen Species in Bovine Somatic Cell Nuclear Transfer Embryos during Micromanipulation Procedures", Reproductive & Developmental biology, 36(1), pp.49~53 (2012), 사단법인 한국동물생명공학회[DOI]
- Im GS, "Development and calcium level changes in pre-implantation porcine nuclear transfer embryos activated with 6-DMAP after fusion", Mol Reprod Dev, 4, pp.1158~1164 (2007)
- 이화연, "Analysis of Endoplasmic Reticulum (ER) Stress Induced during Somatic Cell Nuclear Transfer (SCNT) Process in Porcine SCNT Embryos", 발생과 생식, 22(1), pp.73~83 (2018), 한국발생생물학회
- Niemann H, "Epigenetic reprogramming in embryonic and foetal development upon somatic cell nuclear transfer cloning", Reproduction, 135, pp.151~163 (2008)
- Ozcan U, "Chemical chaperones reduce ER stress and restore glucose homeostasis in a mouse model of type 2 diabetes", Science, 313, pp.1137~1140 (2006)
- 박혜빈, "Endoplasmic Reticulum (ER) Stress and Apoptosis in Parthenogenetic Porcine Embryos following Different Combination of Activation Methods", 한국동물생명공학회지, 32(1), pp.25~31 (2017), 사단법인 한국동물생명공학회
- Park YR, , KSAR & KSET Symposium Proc, pp.29~ (2018)
- Sano R, "ER stress-induced cell death mechanisms", Biochim Biophys Acta-Mol Cell Res, 1833, pp.3460~3470 (2013)
- 송창화, "Endoplasmic Reticulum Stress Responses and Apoptosis", Journal of Bacteriology and Virology, 42(3), pp.196~202 (2012), 대한미생물학회
- Xie Q, "Effect of tauroursodeoxycholic acid on endoplasmic reticulum stress–induced caspase–12 activation", Hepatology, 36, pp.592~601 (2002)
- Yoshida H, "ER stress and diseases", FEBS J, 274, pp.630~658 (2007)
- Zhang JY, "Inhibition of endoplasmic reticulum stress improves mouse embryo development", PLoS ONE, 7, pp.e40433~ (2012)
- Zhang JY, "Effect of endoplasmic reticulum stress on porcine oocyte maturation and parthenogenetic embryonic development in vitro", Biol Reprod, 86, pp.128~ (2011)