earticle

영어

Mitochondria have vital functions that regulate Ca2+ homeostasis and ATP production in fertilized mammalian embryos. In addition, regulation of mitochondrial Ca2+ is related to the mitochondrial functions and mitochondria mediated apoptosis. However, early embryonic development according to the change of mitochondrial Ca2+ regulation during IVF of porcine oocyte has not been reported. In this study, we investigated the regulation of mitochondrial Ca2+ by ruthenium red (RR) during IVF affects blastocyst development in porcine embryos. Here, we investigated the changes of mitochondrial Ca2+ by using Rhod-2 staining in fertilized oocytes during IVF progression (0, 3 and 6 h). After IVF, Rhod-2 expression significantly increased in most of fertilized oocytes at 3 and 6 h. Therefore, we treated RR into IVF medium for reduction of mitochondrial Ca2+. As expected, expressions of rhod-2 and mito-sox significantly decreased in RR treated zygotes. To confirm the mitochondrial functions, we performed the JC-1 and ATP determination in porcine zygotes. As a result, MMP and ATP increased in fertilized oocytes after RR treatment. Interestingly, blastocyst development rate was significantly increased in 20 μM RR treated fertilizing oocytes compared to other groups (p <0.05; 20 μM RR: 33.2±5.0% vs. 10 μM RR: 22.4±5.0% and Con: 24.7±3.8%). In addition, we confirmed that apoptosis in porcine blastocysts was reduced according to RR treatment during IVF procedure by using a TUNEL assay. These results demonstrated that positive effects of RR for regulation of mitochondrial Ca2+ during IVF progression improve the mitochondrial functions and blastocyst developmental competence in porcine embryos.