Opposite regulation of transepithelial electrical resistance and paracellular permeability by Rho in Madin-Darby canine kidney cells.
Small GTPase Rho has been thought to be important for the formation and the maintenance of tight junction in epithelial cells, but the role of Rho in the regulation of barrier function of tight junction is not well understood. We here examined whether Rho was involved in the barrier function of tight junction in Madin-Darby canine kidney (MDCK) cells. The activation of prostaglandin EP3beta receptor, coupled to a Rho activation pathway, induced the increase in transepithelial electrical resistance (TER) but the increase in paracellular flux of mannitol in the preformed monolayer of the MDCK cells expressing the EP3beta receptor. This effect of the EP3 receptor was mimicked by the expression of constitutively active RhoA but not by active Rac1 in MDCK cells, using an isopropyl-beta-D-thiogalactoside-inducible expression system. On the other hand, the activation of EP3beta receptor suppressed the elevation of TER and the decrease in paracellular mannitol flux during Ca(2+) switch-induced tight junction formation, whereas the expression of active RhoA or Rac1 did not apparently affect the TER development in the Ca(2+) switch. These results demonstrate that the EP3 receptor and active RhoA regulate permeabilities of ionic and nonionic molecules in opposite directions in the preformed monolayer, and the EP3 receptor suppresses the elevation of TER during the tight junction formation.
Department of Molecular Neurobiology, Faculty of Pharmaceutical Sciences, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan.
H
H
Hasegawa
H
H
Fujita
H
H
Katoh
J
J
Aoki
K
K
Nakamura
A
A
Ichikawa
M
M
Negishi
D000818:Animals; D001692:Biological Transport / Q000502:physiology; D002460:Cell Line; D002462:Cell Membrane / Q000502:physiology; D002463:Cell Membrane Permeability / Q000502:physiology; D004285:Dogs; D004594:Electrophysiology; D019204:GTP-Binding Proteins / Q000502:physiology*; D007668:Kidney / Q000502:physiology*; D008565:Membrane Proteins / Q000502:physiology*; D018078:Receptors, Prostaglandin E / Q000502:physiology; D020743:rhoB GTP-Binding Protein
D008565:Membrane Proteins; D018078:Receptors, Prostaglandin E; D019204:GTP-Binding Proteins; D020743:rhoB GTP-Binding Protein