| Overexpression of neuronal intermediate filament protein alpha-internexin in PC12 cells. | |
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MedLine Citation:
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PMID: 15880430 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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. |
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Authors:
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Chung-Liang Chien; Tzu-Chiang Liu; Chung-Liang Ho; Kuo-Shyan Lu |
Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Journal of neuroscience research Volume: 80 ISSN: 0360-4012 ISO Abbreviation: J. Neurosci. Res. Publication Date: 2005 Jun |
Date Detail:
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Created Date: 2005-05-25 Completed Date: 2005-08-17 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 7600111 Medline TA: J Neurosci Res Country: United States |
Other Details:
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Languages: eng Pagination: 693-706 Citation Subset: IM |
Copyright Information:
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(c) 2005 Wiley-Liss, Inc. |
Affiliation:
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Department of Anatomy and Cell Biology, College of Medicine, National Taiwan University, Taipei, Taiwan. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals 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 Rats Transfection |
| Chemical | |
Reg. No./Substance:
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0/Carrier Proteins; 0/Intermediate Filament Proteins; 0/alpha-internexin; 147336-22-9/Green Fluorescent Proteins |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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