Tight Junction Dynamics: Oscillations and the Role of Protein Kinase C

Tight Junction Dynamics: Oscillations and the Role of Protein Kinase C The present study aimed to characterize the role of protein kinase C (PKC) on the dynamics of tight junction (TJ) opening and closing in the frog urinary bladder. The early events of TJ dynamics were evaluated by the fast Ca++ switch assay (FCSA), which consisted in opening the TJs by removing basolateral Ca++ ([Ca++] bl ), and closing them by returning [Ca++] bl to normal values. Changes in TJ permeability can be reliably gauged through changes of transepithelial electrical conductance (G) determined in the absence of apical Na+. The FCSA allows the appraisal of drugs and procedures acting upon the mechanism controlling the TJs. The time courses of TJ opening and closing in an FCSA were shown to follow single exponential time courses. PKC inhibition by H7 (100 μm) caused a reduction of the rate of junction opening in response to removing [Ca++] bl , without affecting junction closing, indicating that PKC is a key element in the control of TJ opening dynamics in this preparation. H7 at 250 μm almost completely inhibits TJ opening in response to basolateral Ca++ withdrawal. Subsequent H7 removal caused a prompt inhibition release characterized by a sharp G increase which, however, once started cannot be stopped by H7 reintroduction, Ca++ being necessary to allow TJ recovery. A step rise of apical Ca++ concentration ([Ca++] ap ) causes a reduction of the rate of TJ opening in a FCSA, an effect that is believed to be mediated by apical Ca++ entering the open TJs. The specific condition of having Ca++ only in the apical solution and the TJs located midway between the Ca++ source (apical solution) and the Ca++-binding sites presumably located at the zonula adhaerens, might configure a situation in which a control feedback loop is set up. A rise of [Ca++] ap during the phase of G increase in an FCSA causes a transient recovery of G followed by a subsequent escape phase where G increases again. Oscillations of G also appear in response to a rise of apical Ca++. Both escape and oscillations result from the properties of the TJ regulatory feedback loop. In conclusion, the present results indicate that PKC plays a key role in TJ opening in response to extracellular Ca++ withdrawal without major effect on the reverse process. In addition, PKC inhibition by H7 not only prevents TJ opening in response to basolateral Ca++ removal but induces a prompt blockade of TJ oscillations induced by apical Ca++, oscillations which reappear again when H7 is removed. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png The Journal of Membrane Biology Springer Journals

Tight Junction Dynamics: Oscillations and the Role of Protein Kinase C

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Copyright © Inc. by 2000 Springer-Verlag New York
Life Sciences; Biochemistry, general; Human Physiology
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