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PINK1 is a serine/threonine kinase that is activated under mitochondrial damages. Parkin is phosphorylated at serine 65 located within the ubiquitin-like (UBL) domain by PINK1 and then its E3 ligase activity is activated ( 8, 9). Taken together, our results suggest that VDAC1 monoubiquitination plays important roles in the pathologies of PD by controlling apoptosis.Īmong the PD-associated genes mentioned above, PARK6 encodes PTEN-induced putative kinase1 (PINK1) and PARK2 encodes Parkin, which is a RING/HECT type E3 ubiquitin ligase. Interestingly, Drosophila Parkin T433N, corresponding to human Parkin T415N, fails to rescue the PD-related phenotypes of Parkin-null flies. To further confirm the relevance of our findings in PD, we identify a missense mutation of Parkin discovered in PD patients, T415N, which lacks the ability to induce VDAC1 monoubiquitination but still maintains polyubiquitination. The transgenic flies expressing Drosophila Porin K273R, corresponding to human VDAC1 K274R, show Parkinson disease (PD)-related phenotypes including locomotive dysfunction and degenerated dopaminergic neurons, which are relieved by suppressing MCU and mitochondrial calcium uptake. VDAC1 deficient with polyubiquitination (VDAC1 Poly-KR) hampers mitophagy, but VDAC1 deficient with monoubiquitination (VDAC1 K274R) promotes apoptosis by augmenting the mitochondrial calcium uptake through the mitochondrial calcium uniporter (MCU) channel. Here, we demonstrate that VDAC1 can be either mono- or polyubiquitinated by Parkin in a PINK1-dependent manner. © 2015 Society for Reproduction and Fertility.VDAC1 is a critical substrate of Parkin responsible for the regulation of mitophagy and apoptosis. Taken together, the finding presented here suggest that when the mitochondria are injured, mitochondrial biogenesis and degradation are induced, and that these processes may contribute to the recuperation of oocytes. The relative gene expression of TFAM was furthermore shown to be significantly higher in CCCP-treated oocytes than in untreated oocytes. CCCP treatment was found to increase the Mt number in the modified maturation medium in which mitochondrial degradation was inhibited by MG132, whereas CCCP treatment did not affect the Mt number in the maturation medium lacking MG132. To examine the effects of CCCP treatment of oocytes on the kinetics of mitochondrial DNA copy number (Mt number), COCs treated with 0 or 10 μM CCCP were cultured for 44 h, after which the Mt number was determined by RT-PCR. When the CCCP-treated COCs were cultured further for 44 h in maturation medium, the ATP levels were restored and the parthenogenetic developmental rate of oocytes to the blastocyst stage was comparable with that of untreated COCs. The CCCP treatment was found to significantly reduce ATP content, increase the amount of phosphorylated AMP-activated protein kinase and elevate reactive oxygen species levels in oocytes. Cumulus cell oocyte complexes (COCs) collected from gilt ovaries were treated with 10 μM carbonyl cyanide-m-chlorophenylhydrazone (CCCP a mitochondrial uncoupler) for 2 h. In this study, we investigated the mitochondrial quality control system in porcine oocytes during meiotic maturation.