title: Phospholipase D1 regulates autophagic flux and clearance of α-synuclein aggregates.
abstract: Many neurodegenerative diseases, such as Alzheimer's disease and Parkinson's disease, are characterized by abnormal accumulations of aggregated proteins. Brains in these diseases also show accumulation of autophagic vesicles in the neuronal cytoplasm, suggesting impairment of the autophagic process. As autophagy involves de novo membrane production and vesicle fusion, extensive changes in lipid molecules are necessary. However, the involvement of signaling lipid-modifying enzymes in autophagy and their roles in neurodegenerative diseases are not clear. Using specific inhibitor, we show that loss of phospholipase D1 (PLD1) activity resulted in an accumulation of microtubule-associated protein light chain 3 (LC3), p62, and polyubiquitinated proteins, signs representing malfunction in autophagic flux. Fluorescence and electron microscopic analyses demonstrated impaired fusion of autophagosomes with lysosomes, resulting in accumulation of autophagosomes. Within the cells with impaired autophagic flux, α-synuclein aggregates accumulated in autophagosomes. Knockdown of PLD1 expression using small interfering RNA also resulted in impaired autophagic flux and accumulation of α-synuclein aggregates in autophagosomes. Neuronal toxicity caused by α-synuclein accumulation was rescued by overexpression of PLD1; however, expression of activity-deficient mutant, PLD1-KRM, showed reduced rescue effects. Finally, we demonstrated that both PLD activity and expression levels were reduced in brain tissues of dementia with Lewy bodies (DLB) patients, whereas the amounts of α-synuclein and p62 were increased in the same tissue samples. Collectively, these results suggest that insufficient PLD activity, and therefore, the changes in phospholipid compositions within membranes, might be an important contributor to impaired autophagic process and protein accumulation in Lewy body diseases.
authors.affiliation: 1] Department of Biomedical Science and Technology, Konkuk University, Seoul, Korea [2] IBST, Konkuk University, Seoul, Korea.
authors.forename: E-J
authors.initials: EJ
authors.lastname: Bae
authors.affiliation: 1] IBST, Konkuk University, Seoul, Korea [2] Department of Anatomy, School of Medicine, Konkuk University, Seoul, Korea.
authors.forename: H-J
authors.initials: HJ
authors.lastname: Lee
authors.affiliation: Department of Molecular Biology, Pusan National University, Pusan, Korea.
authors.forename: Y-H
authors.initials: YH
authors.lastname: Jang
authors.affiliation: Department of Neurosciences and Pathology, School of Medicine, University of California San Diego, La Jolla, CA, USA.
authors.forename: S
authors.initials: S
authors.lastname: Michael
authors.affiliation: Department of Neurosciences and Pathology, School of Medicine, University of California San Diego, La Jolla, CA, USA.
authors.forename: E
authors.initials: E
authors.lastname: Masliah
authors.affiliation: Department of Molecular Biology, Pusan National University, Pusan, Korea.
authors.forename: D S
authors.initials: DS
authors.lastname: Min
authors.affiliation: 1] Department of Biomedical Science and Technology, Konkuk University, Seoul, Korea [2] IBST, Konkuk University, Seoul, Korea.
authors.forename: S-J
authors.initials: SJ
authors.lastname: Lee
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D001343:Autophagy; D045744:Cell Line, Tumor; D005260:Female; D006801:Humans; D016631:Lewy Bodies / Q000201:enzymology; D020961:Lewy Body Disease / Q000201:enzymology; D008297:Male; D010300:Parkinson Disease / Q000201:enzymology; D010588:Phagosomes / Q000201:enzymology; D010739:Phospholipase D / Q000502:physiology*; D066329:Protein Aggregates; D051844:alpha-Synuclein / Q000378:metabolism*
chemical_list: D066329:Protein Aggregates; D051844:alpha-Synuclein; D010739:Phospholipase D; C106801:phospholipase D1