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Overexpression of α‐synuclein decreased viability and enhanced sensitivity to prostaglandin E 2 , hydrogen peroxide, and a nitric oxide donor in differentiated neuroblastoma cells
Author(s) -
Prasad Judith E.,
Kumar Bipin,
Andreatta Cynthia,
Nahreini Piruz,
Hanson Amy J.,
Yan Xiang Dong,
Prasad Kedar N.
Publication year - 2004
Publication title -
journal of neuroscience research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.72
H-Index - 160
eISSN - 1097-4547
pISSN - 0360-4012
DOI - 10.1002/jnr.20058
Subject(s) - nitric oxide , neurodegeneration , oxidative stress , viability assay , chemistry , microbiology and biotechnology , cell culture , neuroblastoma , proteasome , biology , cell , biochemistry , endocrinology , medicine , disease , genetics
Increased accumulation of α‐synuclein is associated with certain neurodegenerative diseases including Parkinson's disease (PD) and Alzheimer's disease (AD). One mechanism of α‐synuclein‐induced toxicity involves increased oxidative stress. It was unknown whether neurons overexpressing α‐synuclein would exhibit increased sensitivity to hydrogen peroxide (H 2 O 2 ) or 3‐morpholinosydnonimine (SIN‐1; a nitrous oxide donor). To study this, we developed a murine neuroblastoma (NB) cell line that overexpresses wild‐type human α‐synuclein (NBP2‐PN54) under the control of the cytomegalovirus (CMV) promoter using a retroviral vector. Human α‐synuclein mRNA and protein were readily detectable in NBP2‐PN54 cells. Results showed that differentiated NBP2‐PN54 cells exhibited decreased viability in comparison to differentiated vector (NBP2‐PN1) and parent (NBP2) control cells. These cells also exhibited increased sensitivity to PGE 2 , H 2 O 2 and SIN‐1. Because of involvement of proteasome inhibition in neurodegeneration, we also investigated whether treatment of differentiated NBP2‐PN54 cells with PGE 2 , H 2 O 2 or SIN‐1 inhibits proteasome activity. Results showed that H 2 O 2 and SIN‐1 inhibited proteasome activity, but PGE 2 did not. These results suggest that overexpression of α‐synuclein not only participates directly in degeneration of neurons, but it also increases the vulnerability of neurons to other potential neurotoxins. © 2004 Wiley‐Liss, Inc.