Document Detail


The effect of GIRK2(wv) on neurite growth, protein expression, and viability in the CNS-derived neuronal cell line, CATH.A-differentiated.
MedLine Citation:
PMID:  15953684     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Death occurs in the homozygous mutant mouse weaver among several classes of neuron in cerebellum and ventral midbrain, because these neurons carry a mutation in the G protein-gated inwardly rectifying potassium channel, Girk2. GIRK2 is expressed in all neuronal types killed by wv in cerebellum and midbrain as well as in neurons elsewhere that suffer lesser consequences. GIRK2(wv) affects neurons postnatally, after proliferation, at the time of final differentiation. To assess the impact of GIRK2(wv) on neuronal development and viability, we introduced cDNA encoding wild-type and mutant channels into a variant of a CNS derived catecholamine cell line (Cath.a) known as Cath.a-differentiated. When cultured in serum-free medium, Cath.a-differentiated cells cease proliferation and undergo morphological differentiation, growing long neurites. Cath.a-differentiated cells do not express endogenous Girk channels. Transfection of GIRK2(wv) resulted in the death of Cath.a-differentiated cells, in a cDNA-concentration dependent manner. The highest concentration of Girk2(wv) cDNA caused loss of about half the cells, the next highest concentration one-third, and the least had no effect on viability. However, even the lowest concentration resulted in disruption of neurite outgrowth and reduced the protein products of co-transfected genes. High concentrations of MK801, which prevent Na(+) influx through the mutant channel, prevented death induced by GIRK2(wv). Cell death and disruption of neurite outgrowth were counteracted in GIRK2(wv)-expressing cells by the presence of an unrelated inwardly rectifying potassium channel, Kir2.3. These results are consistent with wv being a gain-of-function mutation, causing disruption of cellular homeostasis by mechanisms such as increased Na(+) influx and chronic depolarization which may in turn result in an excessive metabolic burden on the cell.
Authors:
J C Schein; J K T Wang; S K Roffler-Tarlov
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, N.I.H., Extramural; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Neuroscience     Volume:  134     ISSN:  0306-4522     ISO Abbreviation:  Neuroscience     Publication Date:  2005  
Date Detail:
Created Date:  2005-07-11     Completed Date:  2005-12-02     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  7605074     Medline TA:  Neuroscience     Country:  United States    
Other Details:
Languages:  eng     Pagination:  21-32     Citation Subset:  IM    
Affiliation:
Department of Neuroscience, Tufts University School of Medicine, 136 Harrison Avenue, Boston, MA 02111, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Blotting, Western / methods
Cell Count / methods
Cell Cycle / genetics
Cell Differentiation / physiology
Cell Line
Central Nervous System / cytology*
Dizocilpine Maleate / pharmacology
Excitatory Amino Acid Antagonists / pharmacology
Gene Expression Regulation / physiology*
Green Fluorescent Proteins / biosynthesis
Immunohistochemistry / methods
Mice
Mice, Neurologic Mutants
Neurites / physiology*
Neurons / cytology*,  physiology
Potassium Channels, Inwardly Rectifying / metabolism
Time Factors
Transfection / methods
Grant Support
ID/Acronym/Agency:
NS20181/NS/NINDS NIH HHS
Chemical
Reg. No./Substance:
0/Excitatory Amino Acid Antagonists; 0/Kcnj4 protein, mouse; 0/Potassium Channels, Inwardly Rectifying; 147336-22-9/Green Fluorescent Proteins; 77086-22-7/Dizocilpine Maleate

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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