-
[show abstract]
[hide abstract]
ABSTRACT: Previous work from our laboratory and others has shown that, in some epithelia, the epithelial sodium channel (ENaC) increases its expression during wound healing. In these cases, inhibition of the channel determines a decrease in the healing rate, a result suggesting a role for ENaC in the overall healing process. To understand further this role of ENaC in epithelia, we explored the participation of ENaC in wound healing in four cultured epithelial cell lines selected on the basis of their different embryonic origins, function and modality of healing, i.e., by lamellipodial cell crawling or by actin cable formation. Three of the cell lines (bovine corneal endothelial cells, rabbit corneal epithelial cells and Madin-Darby canine kidney cells) exhibited an increase in ENaC expression and consequent membrane potential depolarization and an increase in cytosolic sodium and calcium, whereas one line (bovine aortal endothelial cells, BAEC) did not exhibit any of these changes. In all of the cell lines, however, ENaC inhibition determined a similar decrease in the rate of wound healing. In BAEC monolayers, the increase in ENaC activity produced plasma membrane depolarization, increased cytosolic sodium and calcium, and augmented the velocity of healing. These novel findings contribute to the idea that ENaC plays a critical role in wound healing in various epithelia, independently of the modality of healing and of any increase in the expression of the channel.
Cell and Tissue Research 05/2013; · 3.11 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Immediately after wounding, bovine corneal endothelial cells develop a fast calcium wave that propagates from the wound border to the rest of the monolayer and extinguishes in approximately 5 minutes. One hour after wounding, a late, slow calcium wave (SCW) develops concomitantly to the depolarization of the plasma membrane potential of the border cells. The incorporation of inhibitors of the epithelial sodium channel and of the sodium-calcium exchanger produces inhibition of the membrane depolarization and the SCW, and diminishes the rate of wound healing. The L-type calcium channel blocker nimodipine does not have any effect on the SCW. The reversible inhibition of the fast calcium wave does not affect the SCW and only slightly decreases the velocity of healing. Our results suggest that the SCW is at least partially produced by the coupling of the epithelial sodium channel and the sodium-calcium exchanger functioning in reverse mode. They also suggest that the SCW may play a role in the overall healing process.
Wound Repair and Regeneration 12/2011; 20(1):28-37. · 2.91 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: We have previously determined that the depolarization of the plasma membrane potential of confluent bovine corneal endothelial cells in culture, provokes a characteristic reorganization of the actin cytoskeleton. The purposes of the present work are to investigate whether similar responses are exhibited by other epithelia, irrespectively of their specific functions and embryonic origin, and to test the hypothesis that the cytoskeletal reorganization induced by membrane depolarization requires of a well-organized circumferential actin disposition in order to take place. For this, we have performed studies on three different cultured epithelia of the eye: bovine corneal endothelium, bovine retinal pigment epithelium and cellular lines of murine lens epithelium. For all of these cells, we explored the effects of plasma membrane depolarization, achieved via the incorporation of gramicidin D to the bathing media or by the replacement of extracellular sodium chloride by potassium gluconate, on the cadherin and actin distribution. The membrane potential changes were monitored by fluorescence microscopy using oxonol V; fluorescent probes were also used for F-actin and cadherin. Detergent extraction and Western blot analysis were employed to reveal the relative amount of cadherin attached to the cytoskeleton. The main findings of this study are that different confluent cultured epithelia exhibiting a well-defined circumferential pattern of actin distribution respond to plasma membrane depolarization by similar modifications in the actin cystoskeleton to those reported for bovine corneal endothelial cells. On the other hand, epithelia that do not exhibit such actin pattern in confluence, as well as non-confluent monolayers, do not display noticeable actin modifications in response to the depolarizing procedures. While in the former cells, cadherin is predominantly located at the lateral membrane domain, the cells that do not respond to membrane depolarization mainly display their cadherin in the intracellular compartment. We suggest that the typical peripheral disposition of actin, associated to well-established epithelial-type adherens junctions (i.e. zonula adherens), is a pre-requisite for the cytoskeletal organizational modifications exhibited by epithelial cells in response to the depolarization of the plasma membrane potential.
Experimental Eye Research 01/2005; 79(6):769-77. · 3.26 Impact Factor
