Document Detail

Overexpression of neuronal intermediate filament protein alpha-internexin in PC12 cells.
MedLine Citation:
PMID:  15880430     Owner:  NLM     Status:  MEDLINE    
The neuronal intermediate filaments include not only the neurofilament triplet proteins but also peripherin and alpha-internexin. To determine whether neurite outgrowth is enhanced by alpha-internexin, the cDNA of rat alpha-internexin tagged with enhanced green fluorescent protein (EGFP) was transfected into a rat adrenal pheochromocytoma cell line PC12 that responds to nerve growth factor (NGF) by induction of the neuronal phenotype. Selected stable clones were induced by NGF and examined for expression patterns of neuronal intermediate filaments by Western blot and immunocytochemistry. Differentiating neurons were also collected after NGF induction for RT-PCR analysis. Overexpressed alpha-internexin-EGFPs were found mainly in cell bodies and the proximal part of neurites. It was also found that overexpression of alpha-internexin-EGFPs enhanced the neurite outgrowth of PC12 cells at the early stages of NGF induction. Meantime, NF-L and NF-M were upregulated by the overexpression of alpha-internexin-EGFPs. Interestingly, alpha-internexin-EGFP-transfected cells obviously detached from culture plates at the later stages of NGF induction. Massive IF accumulations, swelling mitochondria, and degenerating neurites with numerous electron-dense granules were observed ultrastructurally in the alpha-internexin-EGFP-transfected cells. In addition, neuronal death was also characterized positively by the TUNEL assay. These observations may imply that cell death was occurring in alpha-internexin-EGFP-transfected cells. From this study, it could be suggested that alpha-internexin plays an important role in neurite outgrowth and regulates the expression of other neurofilaments during neuronal development. Apoptosis-like cell death could also be induced by the overexpression of alpha-internexin-EGFP in PC12 cells after NGF induction.
Chung-Liang Chien; Tzu-Chiang Liu; Chung-Liang Ho; Kuo-Shyan Lu
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of neuroscience research     Volume:  80     ISSN:  0360-4012     ISO Abbreviation:  J. Neurosci. Res.     Publication Date:  2005 Jun 
Date Detail:
Created Date:  2005-05-25     Completed Date:  2005-08-17     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  7600111     Medline TA:  J Neurosci Res     Country:  United States    
Other Details:
Languages:  eng     Pagination:  693-706     Citation Subset:  IM    
Copyright Information:
(c) 2005 Wiley-Liss, Inc.
Department of Anatomy and Cell Biology, College of Medicine, National Taiwan University, Taipei, Taiwan.
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MeSH Terms
Carrier Proteins / genetics*,  metabolism*
Cell Death / physiology*
DNA Fragmentation
Gene Expression
Green Fluorescent Proteins / genetics
In Situ Nick-End Labeling
Intermediate Filament Proteins
Microscopy, Electron
Neurites / physiology
Neurons / physiology*,  ultrastructure
PC12 Cells
Reg. No./Substance:
0/Carrier Proteins; 0/Intermediate Filament Proteins; 0/alpha-internexin; 147336-22-9/Green Fluorescent Proteins

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