PMID- 29734780 OWN - NLM STAT- MEDLINE DCOM- 20180920 LR - 20220408 IS - 1422-0067 (Electronic) IS - 1422-0067 (Linking) VI - 19 IP - 5 DP - 2018 May 6 TI - cPKCgamma-Modulated Sequential Reactivation of mTOR Inhibited Autophagic Flux in Neurons Exposed to Oxygen Glucose Deprivation/Reperfusion. LID - 10.3390/ijms19051380 [doi] LID - 1380 AB - We have reported that conventional protein kinase Cγ (cPKCγ)-modulated neuron-specific autophagy improved the neurological outcome of mice following ischemic stroke through the Akt-mechanistic target of rapamycin (mTOR) pathway. However, its detailed molecular mechanism remains unclear. In this study, primary cortical neurons from postnatal one-day-old C57BL/6J cPKCγ wild-type (cPKCγ(+/+)) and knockout (cPKCγ(−/−)) mice suffering oxygen glucose deprivation/reperfusion (OGD/R) were used to simulate ischemia/reperfusion injury in vitro. A block of autophagic flux was observed in cPKCγ(+/+) neurons under OGD/R exposure, characterized by accumulation of p62. Immunofluorescent results showed a decrease in colocalization between LC3 and Atg14 or Stx17 in cPKCγ(+/+) neurons when compared with cPKCγ(−/−) neurons after OGD/R. However, the colocalization between LC3 and Lamp2 was barely decreased, indicating the presence of autolysosomes. The larger lysotracker-positive structures were also significantly increased. These results suggest that cPKCγ-induced inhibition of autophagy occurred at the stages of autophagosome formation, Stx17 anchoring, and the degradation of autolysosomes in particular. In addition, cPKCγ-modulated phosphorylation of mTOR at Ser 2481 was dependent on the site of Ser 2448, which may have blocked autophagic flux. cPKCγ-modulated sequential reactivation of mTOR inhibited autophagic flux in neurons exposed to OGD/R, which may provide endogenous interventional strategies for stroke, especially ischemia/reperfusion injury. FAU - Hua, Rongrong AU - Hua R AUID- ORCID: 0000-0001-5102-115X AD - Department of Neurobiology and Center of Stroke, Beijing Institute for Brain Disorders, Capital Medical University, Beijing 100069, China. rrhua@mail.ccmu.edu.cn. FAU - Han, Song AU - Han S AD - Department of Neurobiology and Center of Stroke, Beijing Institute for Brain Disorders, Capital Medical University, Beijing 100069, China. songhan@ccmu.edu.cn. FAU - Zhang, Nan AU - Zhang N AD - Department of Neurobiology and Center of Stroke, Beijing Institute for Brain Disorders, Capital Medical University, Beijing 100069, China. nanzhang@mail.ccmu.edu.cn. FAU - Dai, Qingqing AU - Dai Q AD - Department of Neurobiology and Center of Stroke, Beijing Institute for Brain Disorders, Capital Medical University, Beijing 100069, China. qqingdai@mail.ccmu.edu.cn. FAU - Liu, Ting AU - Liu T AD - Department of Neurobiology and Center of Stroke, Beijing Institute for Brain Disorders, Capital Medical University, Beijing 100069, China. tingl@mail.ccmu.edu.cn. FAU - Li, Junfa AU - Li J AUID- ORCID: 0000-0002-1930-9724 AD - Department of Neurobiology and Center of Stroke, Beijing Institute for Brain Disorders, Capital Medical University, Beijing 100069, China. junfali@ccmu.edu.cn. LA - eng PT - Journal Article DEP - 20180506 PL - Switzerland TA - Int J Mol Sci JT - International journal of molecular sciences JID - 101092791 RN - EC 2.7.1.- (protein kinase C gamma) RN - EC 2.7.1.1 (mTOR protein, mouse) RN - EC 2.7.11.1 (Oncogene Protein v-akt) RN - EC 2.7.11.1 (TOR Serine-Threonine Kinases) RN - EC 2.7.11.13 (Protein Kinase C) RN - IY9XDZ35W2 (Glucose) RN - S88TT14065 (Oxygen) SB - IM MH - Animals MH - Autophagy/drug effects MH - Cell Survival/genetics MH - Cerebral Cortex/metabolism/pathology MH - Glucose/metabolism MH - Mice MH - Neurons/*metabolism/pathology MH - Oncogene Protein v-akt/genetics MH - Oxygen/metabolism MH - Primary Cell Culture MH - Protein Kinase C/*genetics MH - Reperfusion Injury/*metabolism/pathology MH - Stroke/*genetics/metabolism/physiopathology MH - TOR Serine-Threonine Kinases/*genetics PMC - PMC5983661 OTO - NOTNLM OT - autophagy OT - conventional protein kinase Cgamma OT - oxygen glucose deprivation OT - reperfusion COIS- The authors declare no conflict of interest. EDAT- 2018/05/08 06:00 MHDA- 2018/09/21 06:00 PMCR- 2018/05/01 CRDT- 2018/05/09 06:00 PHST- 2018/03/26 00:00 [received] PHST- 2018/04/24 00:00 [revised] PHST- 2018/05/03 00:00 [accepted] PHST- 2018/05/09 06:00 [entrez] PHST- 2018/05/08 06:00 [pubmed] PHST- 2018/09/21 06:00 [medline] PHST- 2018/05/01 00:00 [pmc-release] AID - ijms19051380 [pii] AID - ijms-19-01380 [pii] AID - 10.3390/ijms19051380 [doi] PST - epublish SO - Int J Mol Sci. 2018 May 6;19(5):1380. doi: 10.3390/ijms19051380.