TRPC1 protects human SH-SY5Y cells against salsolinol-induced cytotoxicity by inhibiting apoptosis.

Abstract

Salsolinol, an endogenous neurotoxin, may be involved in the pathogenesis of Parkinson's disease. In this study, we sought to determine whether salsolinol-induced cytotoxicity in SH-SY5Y human neuroblastoma cells, a cloned cell line which expresses dopaminergic activity, could be prevented by overexpressing a Ca(2+) channel, transient receptor potential (TRPC1) protein. Exposure of SH-SY5Y cells to 500 microM salsolinol for 12 h resulted in a significant decrease in thapsigargin or carbachol-mediated Ca(2+) influx. Consistent with these results, SH-SY5Y cells treated with salsolinol showed approximately 60% reduction in TRPC1 protein levels. Confocal microscopy also showed that SH-SY5Y cells treated with salsolinol had a significant decrease in the plasma membrane staining of the TRPC1 protein. Interestingly, overexpression of TRPC1 increases TRPC1 protein levels and also protected SH-SY5Y neuroblastoma cells against salsolinol-mediated cytotoxicity as determined by 3,[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay. The protective effect of TRPC1 was blocked by the addition of TRPC1 blockers lanthanum, or 2APB. Activation of TRPC1 protein by either thapsigargin or carbachol further protected SH-SY5Y cells from salsolinol treatments. Staining of SH-SY5Y cells with an apoptotic marker (YO-PRO-1) showed that TRPC1 protein protects against apoptosis. Furthermore, TRPC1 overexpression also inhibited cytochrome c release and decreased BAX protein levels required for apoptosis. Taken together, these findings suggest that the reduction in cell surface TRPC1 protein expression in response to salsolinol may be a contributory factor in cellular toxicity of the dopaminergic neurons. Furthermore, overexpression of TRPC1 could inhibit apoptotic complex thereby increasing neuronal cell survivability in Parkinson's disease.

• Authors: Sunitha Bollimuntha, Manuchair Ebadi, Brij B Singh
• Journal: Brain research (Brain Res) Vol. 1099 Issue 1 Pg. 141-9 (Jul 12 2006) ISSN: 0006-8993 [Print] Netherlands
• PMID: 16765919 (Publication Type: Comparative Study, Journal Article, Research Support, N.I.H., Extramural, Research Support, U.S. Gov't, Non-P.H.S.)

Chemical References

• APAF1 protein, human
• Apoptotic Protease-Activating Factor 1
• Intracellular Signaling Peptides and Proteins
• Isoquinolines
• Proteins
• TRPC Cation Channels
• Tetrazolium Salts
• Thiazoles
• transient receptor potential cation channel, subfamily C, member 1
• salsolinol
• thiazolyl blue
• Calcium

Topics

• Apoptosis (drug effects)
• Apoptotic Protease-Activating Factor 1
• Blotting, Western (methods)
• Calcium (metabolism)
• Cell Count (methods)
• Cell Line, Tumor
• Gene Expression (drug effects)
• Humans
• Intracellular Signaling Peptides and Proteins (metabolism)
• Isoquinolines (toxicity)
• Microscopy, Confocal (methods)
• Neuroblastoma (pathology)
• Proteins (metabolism)
• TRPC Cation Channels (metabolism)
• Tetrazolium Salts
• Thiazoles
• Time Factors