Park et al. (2014). Neurobiology of Aging. doi: 10.1016/j.neurobiolaging.2014.01.028
…e incubated overnight at 4 C with primary antibody: mouse anti-tyrosine hydroxylase (TH, 1:2000 dilution for brain tissue, Pel-freez, St. Arkansas, AR, USA). Following this, the brain sections and the cells were rinsed 3 times in 0.5% BSA in 1 PBS (10 min/rinse) and incubated with the appropriate biotinylated secondary antibody and avidinebiotin complex (Elite Kit; Vector Laboratories, Burlin
Abstract
Autophagy is a major degradation pathway for abnormal aggregated proteins and organelles that cause various neurodegenerative diseases. Current evidence suggests a central role for autophagy in pathogenesis of Parkinson's disease, and that dysfunction in the autophagic system may lead to α-synuclein accumulation. In the present study, we investigated whether mesenchymal stem cells (MSCs) would enhance autophagy and thus exert a neuroprotective effect through the modulation of α-synuclein in parkinsonian models. In MPP(+)-treated neuronal cells, coculture with MSCs increased cellular viability, attenuated expression of α-synuclein, and enhanced the number of LC3-II-positive autophagosomes compared with cells treated with MPP(+) only. In an MPTP-treated animal model of Parkinson's disease, MSC administration significantly increased final maturation of late autophagic vacuoles, fusion with lysosomes. Moreover, MSC administration significantly reduced the level of α-synuclein in dopaminergic neurons, which was elevated in MPTP-treated mice. These results suggest that MSC treatment significantly enhances autophagolysosome formation and may modulate α-synuclein expression in parkinsonian models, which may lead to increased neuronal survival in the presence of neurotoxins.