Document Detail


C-terminal tagging with eGFP yields new insights into expression of connexin45 but prevents rescue of embryonic lethal connexin45-deficient mice.
MedLine Citation:
PMID:  19473725     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Connexin45 (Cx45) is a member of the connexin family which can form gap junction channels and is known to be expressed in several cell types in the embryonic as well as adult mouse including working cardiomyocytes and certain types of neurons. Until now its subcellular localization could not be unequivocally determined in certain tissues due to the lack of sensitive and specific antibodies. In order to investigate the localization of Cx45, we have generated a transgenic mouse expressing a fusion protein composed of Cx45 and eGFP under control of the endogenous Cx45 promoter using a bacterial artificial chromosome (BAC). In previous studies it had been shown that a C-terminal tag of connexin proteins only slightly altered the properties of gap junction channels in cultured cells and allowed direct visualization of the fusion protein. In the adult brain the expression of the Cx45eGFP protein was found in the subventricular zone in transient amplifying cells as well as in neuroblasts and ependymal cells. In addition Cx45eGFP is expressed in the atrial and ventricular working myocardium, i.e. regions of the heart where divergent results regarding Cx45 expression had previously been published. In the lung we identified Cx45eGFP in the smooth muscle cell layer of bronchioles. The Cx45eGFP transgene could not rescue embryonic lethality of Cx45-deficient mice, i.e. Cx45eGFP//Cx45(-/-) mice die around ED10.5 presumably due to altered properties of gap junction channels as a result of C-terminal tagging of Cx45.
Authors:
Julia von Maltzahn; Maria M Kreuzberg; Gabi Matern; Carsten Euwens; Thorsten H?her; Philipp W?rsd?rfer; Klaus Willecke
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2009-05-27
Journal Detail:
Title:  European journal of cell biology     Volume:  88     ISSN:  1618-1298     ISO Abbreviation:  Eur. J. Cell Biol.     Publication Date:  2009 Sep 
Date Detail:
Created Date:  2009-07-20     Completed Date:  2009-12-15     Revised Date:  2010-05-26    
Medline Journal Info:
Nlm Unique ID:  7906240     Medline TA:  Eur J Cell Biol     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  481-94     Citation Subset:  IM    
Affiliation:
Institute of Genetics, Division of Molecular Genetics, University of Bonn, Bonn, Germany.
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MeSH Terms
Descriptor/Qualifier:
Animals
Chromosomes, Artificial, Bacterial / genetics,  metabolism
Connexins / genetics*,  metabolism*
Fluorescent Antibody Technique
Genes, Lethal
Genotype
Green Fluorescent Proteins / genetics*,  metabolism
Mice
Mice, Transgenic
Chemical
Reg. No./Substance:
0/Connexins; 0/connexin 45; 0/enhanced green fluorescent protein; 147336-22-9/Green Fluorescent Proteins

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