A neuronal death model: overexpression of neuronal intermediate filament protein peripherin in PC12 cells

Journal of Biomedical Science

Research

Daphne Kan¹  Wen-Ching Lee²  Yun-Yu Chen²  Chung-Liang Chien³ 

[1] Center of Genomic Medicine, National Taiwan University, Jen-Ai Road, 100, Taipei, Taiwan;Department of Anatomy and Cell Biology, College of Medicine, National Taiwan University, Jen-Ai Road, 100, Taipei, Taiwan;Department of Anatomy and Cell Biology, College of Medicine, National Taiwan University, Jen-Ai Road, 100, Taipei, Taiwan;Center of Genomic Medicine, National Taiwan University, Jen-Ai Road, 100, Taipei, Taiwan;

关键词: Amyotrophic Lateral Sclerosis;    PC12 Cell;    Nerve Growth Factor;    Intermediate Filament;    Amyotrophic Lateral Sclerosis Patient;   

DOI  :  10.1186/1423-0127-19-8

 received in 2011-11-08, accepted in 2012-01-17,  发布年份 2012

来源: Springer

【 摘 要 】

BackgroundAbnormal accumulation of neuronal intermediate filament (IF) is a pathological indicator of some neurodegenerative disorders. However, the underlying neuropathological mechanisms of neuronal IF accumulation remain unclear. A stable clone established from PC12 cells overexpressing a GFP-Peripherin fusion protein (pEGFP-Peripherin) was constructed for determining the pathway involved in neurodegeneration by biochemical, cell biology, and electronic microscopy approaches. In addition, pharmacological approaches to preventing neuronal death were also examined.ResultsResults of this study showed that TUNEL positive reaction could be detected in pEGFP-Peripherin cells. Swollen mitochondria and endoplasmic reticulum (ER) were seen by electron microscopy in pEGFP-Peripherin cells on day 8 of nerve growth factor (NGF) treatment. Peripherin overexpression not only led to the formation of neuronal IF aggregate but also causes aberrant neuronal IF phosphorylation and mislocation. Western blots showed that calpain, caspase-12, caspase-9, and caspase-3 activity was upregulated. Furthermore, treatment with calpain inhibitor significantly inhibited cell death.ConclusionsThese results suggested that the cytoplasmic neuronal IF aggregate caused by peripherin overexpression may induce aberrant neuronal IF phosphorylation and mislocation subsequently trapped and indirectly damaged mitochondria and ER. We suggested that the activation of calpain, caspase-12, caspase-9, and caspase-3 were correlated to the dysfunction of the ER and mitochondria in our pEGFP-Peripherin cell model. The present study suggested that pEGFP-Peripherin cell clones could be a neuronal death model for future studies in neuronal IFs aggregate associated neurodegeneration.

【 授权许可】

Unknown   
© Lee et al; licensee BioMed Central Ltd. 2012. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.