KMID : 0894520190230010043
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Development & Reproduction 2019 Volume.23 No. 1 p.43 ~ p.54
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Effects of Endoplasmic Reticulum Stress Inhibitor Treatment during the Micromanipulation of Somatic Cell Nuclear Transfer in Porcine Oocytes
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Park Yeo-Reum
Park Hye-Bin Kim Mi-Jeong Jung Bae-Dong Lee Seung-Hyung Park Choon-Keun Cheong Hee-Tae
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Abstract
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We examined the effects of endoplasmic reticulum (ER) stress inhibitor treatment during the micromanipula-tion of porcine somatic cell nuclear transfer (SCNT) on the in vitro development of SCNT embryos. ER stress inhibitors such as salubrinal (200 nM) and tauroursodeoxycholic acid (TUDCA; 100 ¥ìM) were added to the micromanipulation medium and holding medium. The expression of X-box binding protein 1 (Xbp1), ER-stress-associated genes, and apoptotic genes in SCNT embryos was confirmed at the one-cell and blastocyst stages. Levels of Xbp1 splicing and expression of ER-stress-associated genes in SCNT embryos at the one-cell stage decreased significantly with TUDCA treatment (p<0.05). The expression of ER-stress-associated genes also decreased slightly with the addition of both salubrinal and TUDCA (Sal+TUD). The expression levels of caspase-3 and Bcl2-associated X protein (Bax) mRNA were also significantly lower in the TUDCA and Sal+TUD treatments (p<0.05). At the blastocyst stage, there were no differences in levels of Xbp1 splicing, and transcription of ER-stress-associated genes and apoptosis genes between control and treatment groups. However, the blastocyst formation rate (20.2%) and mean blastocyst cell number (63.0¡¾7.2) were significantly higher (p<0.05) for embryos in the TUDCA treatment compared with those for control (12.6% and 41.7¡¾3.1, respectively). These results indicate that the addition of ER-stress inhibitors, especially TUDCA, during micromanipulation can inhibit cellular damage and enhance in vitro development of SCNT embryos by reducing stress levels in the ER.
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KEYWORD
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Somatic cell nuclear transfer, Endoplasmic reticulum (ER) stress inhibitor, Micromanipulation, Apoptosis, Porcine
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