Regulation of epithelial apical junctions and barrier function by Galpha13.
The epithelial tight junction forms a barrier to paracellular solute movement. In this study we show that the heterotrimeric G-protein Galpha13 regulates the epithelial tight junction barrier. We generated MDCKII kidney epithelial cell lines in which the expression of an active Galpha13 mutant (Galpha13Q226L) could be induced. We demonstrated that Galpha13Q226L expression increased paracellular permeability and caused the disruption and redistribution of proteins comprising the tight junction and the adherens junction away from sites of cell contact and the appearance of basal stress fibers. The effects on the junctional proteins and the actin cytoskeleton were abrogated by the Rho kinase inhibitor Y27632 but not by the Src kinase inhibitor PP2. The Galpha13 mediated increase in permeability was also Src kinase independent but was partly dependent on Rho kinase signalling. Our data establish a link between Galpha13, Rho kinase signaling and epithelial barrier function and not only demonstrate that Galpha13 regulates epithelial apical junction properties but that it does so via signaling pathways that are distinct from the closely related protein Galpha12.
Donato
Rino
R
Women's and Children's Health Research Institute, North Adelaide, South Australia, Australia.
Wood
Stephen A
SA
Saunders
Ian
I
Gundsambuu
Batjargal
B
Yan Mak
Kai
K
Abbott
Catherine A
CA
Powell
Barry C
BC
D022005:Adherens Junctions / Q000502:physiology*; D000577:Amides / Q000494:pharmacology; D000818:Animals; D002463:Cell Membrane Permeability / Q000502:physiology; D004285:Dogs; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004848:Epithelium / Q000378:metabolism*; D044365:GTP-Binding Protein alpha Subunits, G12-G13 / Q000502:physiology*; D007124:Immunoenzyme Techniques; D007668:Kidney / Q000166:cytology / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D011725:Pyridines / Q000494:pharmacology; D019108:Tight Junctions / Q000502:physiology*; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D019061:src-Family Kinases / Q000378:metabolism*
D000577:Amides; D004791:Enzyme Inhibitors; D011725:Pyridines; C108830:Y 27632; D019061:src-Family Kinases; D054460:rho-Associated Kinases; D044365:GTP-Binding Protein alpha Subunits, G12-G13