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Alteration in Glutathione Content and Associated Enzyme Activities in the Synaptic Terminals but not in the Non-synaptic Mitochondria from the Frontal Cortex of Parkinson’s Disease Brains

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Abstract

Altered redox dynamics contribute to physiological aging and Parkinson’s disease (PD). This is reflected in the substantia nigra (SN) of PD patients as lowered antioxidant levels and elevated oxidative damage. Contrary to this observation, we previously reported that non-SN regions such as caudate nucleus and frontal cortex (FC) exhibited elevated antioxidants and lowered mitochondrial and oxidative damage indicating constitutive protective mechanisms in PD brains. To investigate whether the sub-cellular distribution of antioxidants could contribute to these protective effects, we examined the distribution of antioxidant/oxidant markers in the neuropil fractions [synaptosomes, non-synaptic mitochondria and cytosol] of FC from PD (n = 9) and controls (n = 8). In the control FC, all the antioxidant activities [Superoxide dismutase (SOD), glutathione (GSH), GSH peroxidase (GPx), GSH-S-transferase (GST)] except glutathione reductase (GR) were the highest in cytosol, but several fold lower in mitochondria and much lower in synaptosomes. However, FC synaptosomes from PD brains had significantly higher levels of GSH (p = 0.01) and related enzymes [GPx (p = 0.02), GR (p = 0.06), GST (p = 0.0001)] compared to controls. Conversely, mitochondria from the FC of PD cases displayed elevated SOD activity (p = 0.02) while the GSH and related enzymes were relatively unaltered. These changes in the neuropil fractions were associated with unchanged or lowered oxidative damage. Further, the mitochondrial content in the synaptosomes of both PD and control brains was ≥five-fold lower compared to the non-synaptic mitochondrial fraction. Altered distribution of oxidant/antioxidant markers in the neuropil fractions of the human brain during aging and PD has implications for (1) degenerative and protective mechanisms (2) distinct antioxidant mechanisms in synaptic terminals compared to other compartments.

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Abbreviations

PD:

Parkinson’s disease

SN:

Substantianigra

FC:

Frontal cortex

GSH:

Glutathione reduced

PMI:

Postmortem interval

3-NT:

3-nitrotyrosine

GFAP:

Glial fibrillary acidic protein

SOD:

Superoxide Dismutase

GST:

Glutathione-s-transferase

GR:

Glutathione reductase

GP:

Xglutathione peroxidase

ns mito:

Non-synaptic mitochondria

syn mito:

Synaptic mitochondria

cyto:

Cytosol

CS:

Citrate synthase

MDH:

Malate dehydrogenase

SDH:

Succinate dehydrogenase

MTT:

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

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Acknowledgments

This work was financially supported by a research grant from the Indian Council of Medical Research (ICMR IRIS ID No. 2009-07710), India, to MMSB. GH is a senior research fellow of ICMR, India. We thank all the donors and their relatives for donating human brain tissue samples for this study. Human brain tissues for the study were sourced from Human Brain Tissue Repository for Neurobiological Studies (A National Research Facility), Department of Neuropathology, NIMHANS, Bangalore–560 029. We thank Dr. Uday Muthane for his help regarding neurological analysis of the PD samples used in the current study. GH and MMSB designed the study. GH carried out the biochemical work. SKS and AM provided the tissue samples and carried out the neuropathological analysis. MMSB and GH analyzed the data. MMSB wrote the paper, while the co-authors assisted in editing it.

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Harish, G., Mahadevan, A., Srinivas Bharath, M.M. et al. Alteration in Glutathione Content and Associated Enzyme Activities in the Synaptic Terminals but not in the Non-synaptic Mitochondria from the Frontal Cortex of Parkinson’s Disease Brains. Neurochem Res 38, 186–200 (2013). https://doi.org/10.1007/s11064-012-0907-x

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