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Overexpression of neurofilament H disrupts normal cell structure and functionStudying exogenously expressed tagged proteins in live cells has become a standard technique for evaluating protein distribution and function. Typically, expression levels of experimentally introduced proteins are not regulated, and high levels are often preferred to facilitate detection. However, overexpression of many proteins leads to mislocalization and pathologies. Therefore, for normative studies, moderate levels of expression may be more suitable. To understand better the dynamics of intermediate filament formation, transport, and stability in a healthy, living cell, we inserted neurofilament heavy chain (NFH)-green fluorescent protein (GFP) fusion constructs in adenoviral vectors with tetracycline (tet)-regulated promoters. This system allows for turning on or off the synthesis of NFH-GFP at a selected time, for a defined period, in a dose-dependent manner. We used this inducible system for live cell imaging of changes in filament structure and cell shape, motility, and transport associated with increasing NFH-GFP expression. Cells with low to intermediate levels of NFH-GFP were structurally and functionally similar to neighboring, nonexpressing cells. In contrast, overexpression led to pathological alterations in both filament organization and cell function. Copyright 2002 Wiley-Liss, Inc.
Document ID
20040088383
Document Type
Reprint (Version printed in journal)
External Source(s)
Authors
Szebenyi, Gyorgyi (University of Texas Southwestern Medical Center 5323 Harry Hines Boulevard, Dallas, TX 75390-9039, United States)
Smith, George M.
Li, Ping
Brady, Scott T.
Date Acquired
August 21, 2013
Publication Date
April 15, 2002
Publication Information
Publication: Journal of neuroscience research
Volume: 68
Issue: 2
ISSN: 0360-4012
Subject Category
Life Sciences (General)
Funding Number(s)
CONTRACT_GRANT: NS 33776
CONTRACT_GRANT: NS 41170-01
CONTRACT_GRANT: NS 38126
CONTRACT_GRANT: NS 23868
CONTRACT_GRANT: AG 12646
CONTRACT_GRANT: NS 23320
Distribution Limits
Public
Copyright
Other
Keywords
Non-NASA Center
NASA Discipline Neuroscience