Effect of Trehalose on PC12 Cells Overexpressing Wild-Type or A53T Mutant α-synuclein

Abstract

Accumulation of α-synuclein (α-Syn) is a common pathology for both familiar and sporadic Parkinson's disease (PD), enhancing its clearance might be a promising strategy for treating PD. To assess the potential of trehalose in this regard, we investigated its effect on the PC12 cells overexpressing wild type (WT) or A53T mutant α-Syn and the implicated pathway it might mediated. We observed that trehalose promoted the clearance of A53T α-Syn but not WT α-Syn in PC12 cells, and confirmed the increased LC3 and Lysotracker RED positive autolysosomes by using lysotracker and LC3 staining, the enhanced expression of LC3-II in Western blot, and more autophagosomes under Transmission Electron Microscope in a dose dependent manner after the trehalose treatment. The activation of autophagy can be alleviated by applying macroautophagy inhibitor 3-methyladenine (3-MA). In addition, degradation of A53T and WT α-Syn was blocked after Ubiquitin Proteasome System (UPS) inhibitor (MG132) was applied in those PC12 cells overexpressing A53T or WT α-Syn, suggesting that A53T α-Syn could be degraded by both UPS and macroautophagy. But the effect of trehalose on A53T α-Syn is mainly mediated through the macroautophagy pathway, which is not a dominant way for WT α-Syn clearance. Further in vivo research will be needed to verify the effectiveness of trehalose in treating PD.

Treatment with Trehalose Prevents Behavioral and Neurochemical Deficits Produced in an AAV α-Synuclein Rat Model of Parkinson’s Disease

“…Our previous work reported that trehalose was effective in disaggregating mutant α-synuclein protofibrils and inhibiting the formation of mutant α-synuclein fibrils in the cell-free system of A53T α-synuclein incubation [30]. Further research confirmed that trehalose could enhance clearance of α-synuclein aggregates through activation of the autophagy pathway [31]. Recently, it has been argued that, given advances in animal Fig.…”

Involvement of mortalin/GRP75/mthsp70 in the mitochondrial impairments induced by A53T mutant α-synuclein

“…The human SH-SY5Y neuroblastoma cells possess many characteristics of DAergic neurons, and in the present study SH-SY5Y cells overexpressing A53T α-syn (SH-SY5Y-A53T) were generated for PD research as described previously (Lan et al, 2012). The cDNA for human mortalin (Yang et al, 2011b) was kindly provided by Professor Wen Liu (Department of Cellular and Genetic Medicine, Fudan University, Shanghai, China), subcloned into pLenti6 vector (Invitrogen, USA) and packaged into viral particles according to the manufacturer's instructions (Biowit Technologies, China).…”

“…After two washes, cells were incubated with secondary antibody (1:100, FITC goat antirabbit IgG, Kangcheng, China) (Qu et al, 2012). A laser scanning microscope (Olympus BX60) was then applied to capture images (Lan et al, 2012).…”

“…Although the mechanisms underlying PD remain elusive, α-synuclein (α-syn) accumulation and mitochondrial dysfunction have been recognized as major contributors (Devi et al, 2008;Lan et al, 2012). Numerous studies suggest a key role for the protein α-syn in the pathogenesis of PD.…”

Anhydrobiosis: An Unsolved Problem with Applications in Human Welfare

“…The cells showed depressed levels of tau aggregation and much less cytotoxicity. In a related study, Lan et al (2012) showed that production of synuclein (a protein related to Parkinson's disease) was suppressed by trehalose induced autophagy. The authors of both these studies suggested that trehalose might be developed as a therapeutic agent for these diseases, but it is, of course, unclear how such a therapy could be developed for clinical application.…”