Reduced expression of peroxisome-proliferator activated receptor gamma coactivator-1alpha enhances alpha-synuclein oligomerization and down regulates AKT/GSK3beta signaling pathway in human neuronal cells that inducibly express alpha-synuclein.
Intracellular accumulation of filamentous alpha-synuclein (alpha-Syn) aggregates to form Lewy bodies is a pathologic hallmark of Parkinson's disease. To determine whether mitochondrial impairment plays a role in the accumulation of alpha-Syn oligomer, we used 3D5 cell culture model of human neuronal type whereby conditional overexpression of wild-type alpha-Syn via the tetracycline-off (TetOff) induction mechanism results in formation of inclusions that exhibit many characteristics of Lewy bodies. In the present study, we compromised mitochondrial function in 3D5 cells by using shRNA to knockdown peroxisome-proliferator activated receptor gamma coactivator-1alpha (PGC-1alpha), a key regulator of mitochondrial biogenesis and cellular energy metabolism and found that PGC-1alpha suppression at both protein and mRNA levels results in alpha-Syn accumulation (i.e. monomeric and oligomeric species in the TetOff-induced cells and monomeric only in the non-induced). These changes were accompanied with reduced mitochondrial potential as well as decreased levels of AKT, GSK3beta (total and Ser(9)-phosphorylated) and p53 that are important for cell survival. The extent to which these proteins decreased following PGC-1alpha knockdown, in contrast to what was demonstrable with the viability assay, is greater in the induced than the non-induced. Together these findings indicate that such knockdown increases the propensity to accumulate alpha-Syn oligomers, but the accumulation appears to have very little toxic impact to the neuronal cells.
Ebrahim
Abdul Shukkur
AS
Department of Neuroscience, Mayo Clinic, 4500 San Pablo Rd., Jacksonville, FL 32224, USA.
Ko
Li-Wen
LW
Yen
Shu-Hui
SH
D001704:Biopolymers; D002460:Cell Line; D015536:Down-Regulation; D055785:Gene Knockdown Techniques; D038362:Glycogen Synthase Kinase 3 / Q000096:biosynthesis*; D000071679:Glycogen Synthase Kinase 3 beta; D006360:Heat-Shock Proteins / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D016631:Lewy Bodies / Q000378:metabolism; D053078:Membrane Potential, Mitochondrial; D008928:Mitochondria / Q000502:physiology; D009474:Neurons / Q000378:metabolism*; D051058:Oncogene Protein v-akt / Q000096:biosynthesis*; D000071248:Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha; D010766:Phosphorylation; D015398:Signal Transduction; D014157:Transcription Factors / Q000096:biosynthesis* / Q000235:genetics; D051844:alpha-Synuclein / Q000378:metabolism*
D001704:Biopolymers; D006360:Heat-Shock Proteins; C486497:PPARGC1A protein, human; D000071248:Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha; D014157:Transcription Factors; D051844:alpha-Synuclein; C000605506:GSK3B protein, human; D000071679:Glycogen Synthase Kinase 3 beta; D051058:Oncogene Protein v-akt; D038362:Glycogen Synthase Kinase 3