Cell-cell interactions in the kidney: inducible expression of mutant G protein alpha-subunits in Madin-Darby canine kidney cells for studies of epithelial cell junction structure and function.
Methods Mol Biol
; 341: 61-72, 2006.
Article
en En
| MEDLINE
| ID: mdl-16799189
ABSTRACT
Disrupted epithelial cell junctions are a hallmark of numerous disease processes. The signaling mechanisms regulating barrier function and re-establishment of intact junctions after injury and during development are complex and tightly regulated. We have shown that heterotrimeric G proteins regulate assembly and maintenance of epithelial cell barrier in cultured Madin-Darby canine kidney (MDCK) cells. The inducible expression of mutant signaling molecules (constitutively active or dominant negative) in polarized cells is a useful strategy for elucidating the role(s) for specific proteins. Using tetracycline-off inducible expression of wild-type and constitutively active Galpha12, we have demonstrated a fundamental role for Galpha12 in regulating the junction of MDCK cells. Inducible expression permits the comparison of the identical cell line in the presence and absence of the protein of interest and minimizes variation arising from distinct clones. The methods described here are applicable to virtually any protein that may regulate maintenance or assembly of the epithelial cell junctional complex.
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Bases de datos:
MEDLINE
Asunto principal:
Transducción de Señal
/
Uniones Estrechas
/
Células Epiteliales
/
Subunidad alfa de la Proteína de Unión al GTP Gi2
/
Riñón
/
Mutación
Límite:
Animals
Idioma:
En
Revista:
Methods Mol Biol
Asunto de la revista:
BIOLOGIA MOLECULAR
Año:
2006
Tipo del documento:
Article
País de afiliación:
Estados Unidos