Rabbit Anti-Human DNM1 (Phospho-Ser778) polyclonal antibody

The purpose of this study was to investigate the changes in mitochondria in porcine MII-stage oocytes after open pulled straw (UPS) vitrification and to determine their roles in apoptosis and in vitro developmental ability. The mitochondrial membrane potential (Delta Psi m), reactive oxygen species (ROS) level, adenosine-5'-triphosphate (ATP) concentration, mitochondrial distribution, mitochondrial ultrastructure, early-stage apoptosis with Annexin V-FITC staining, survival rate, parthenogenetic developmental ability and related gene expression were measured in the present experiments. The results showed that: (1) the mitochondrial Delta Psi m of vitrified-thawed oocytes (1.05) was lower than that of fresh oocytes 1.24 (P < 0.05). (2) ROS level in the UPS vitrification group was much higher than that of the fresh group, while the ATP concentration was much lower than that of fresh group (P < 0.05). (3) Early-stage apoptosis rate from the UPS vitrification group (57.6%) was much higher than that of fresh group (8.53%) (P < 0.05), and the survival rate and parthenogenetic cleavage rate of UPS vitrified oocytes were much lower than those from fresh ones (P < 0.05). (4) Vitrification not only disrupted the mitochondrial distribution of porcine MII-stage oocytes, but also damaged the mitochondrial ultrastructure. (5) After vitrification, the gene expression level of Dnm1 was up-regulated, and other four genes (SOD1, Mfn2, BAX and Bcl2) were down-regulated. The present study suggested that not only the morphology and function of mitochondria were damaged greatly during the vitrification process, but also early-stage apoptosis was observed after vitrification. Intrinsic mitochondrial pathway could be in involved in the occurrence of apoptosis in vitrified-thawed porcine oocytes. (C) 2015 Elsevier Inc. All rights reserved.

In yeast, C-tail-anchored mitochondrial outer membrane protein Fis1 recruits the mitochondrial-fission-regulating GTPase Dnm1 to mitochondrial fission sites. However, the function of its mammalian homologue remains enigmatic because it has been reported to be dispensable for the mitochondrial recruitment of Drp1, a mammalian homologue of Dnm1. We identified TBC1D15 as a Fis1-binding protein in HeLa cell extracts. Immunoprecipitation revealed that Fis1 efficiently interacts with TBC1D15 but not with Drp1. Bacterially expressed Fis1 and TBC1D15 formed a direct and stable complex. Exogenously expressed TBC1D15 localized mainly in cytoplasm in HeLa cells, but when coexpressed with Fis1 it localized to mitochondria. Knockdown of TBC1D15 induced highly developed mitochondrial network structures similar to the effect of Fis1 knockdown, suggesting that the TBC1D15 and Fis1 are associated with the regulation of mitochondrial morphology independently of Drp1. These data suggest that Fis1 acts as a mitochondrial receptor in the recruitment of mitochondrial morphology protein in mammalian cells.