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Evidence for a Na+/Ca2+ exchange mechanism in frog skin epithelium
Authors:Katja H. Madsen  Birger Brodin  R. Nielsen
Affiliation:(1) Biochemical Department, August Krogh Institute, Universitetsparken 13, DK-2100 Copenhagen, Denmark (e-mail: RNielsen@aki.ku.dk), DK;(2) Royal Danish School of Pharmacy, Universitetsparken 2, DK-2100 Copenhagen, Denmark, DK
Abstract: In the present study we investigated the possible existence of a Na+/Ca2+ exchange mechanism in the basolateral membrane of the frog skin epithelium and whether such a mechanism plays a role in the regulation of transepithelial Na+ transport. Cytosolic calcium ([Ca2+]i) was measured with the probe fura-2 in a set-up in which pieces of tissue were mounted on the stage of an epifluorescence microscope. Na+ transport was measured as the amiloride-sensitive short-circuit current (I sc) using a conventional voltage clamp. Basal [Ca2+]i was 65±6 nM (n=15). Removal of Na+ from the mucosal solution had no effect on [Ca2+]i. When Na+ was removed from the serosal solution, [Ca2+]i increased biphasically to a peak of 220±38 nM (n=8, P=0.006). Readdition of Na+ to the serosal solution returned [Ca2+]i to control level. The serosal Na+ gradient and changes in [Ca2+]i were closely correlated; stepwise changes in serosal Na+ were followed by stepwise changes in [Ca2+]i. These observations indicate the existence of a Na+/Ca2+ exchange mechanism in the basolateral membrane of the frog skin epithelium. The transepithelial Na+ transport decreased from 13.2±1.8 to 9.2±1.5 μA cm–2 (n=8, P=0.049) when Na+ was omitted from the serosal solution. When this protocol was repeated in the absence of serosal Ca2+, Na+ transport decreased similarly from 16.7±1.7 to 11.6 ±1.8 μA cm–2 (n=6, P=0.004). We conclude that it is unlikely that the observed decrease in I sc after removal of serosal Na+ is due to an increase in [Ca2+]i per se. Received: 10 July 1998 / Received after revision: 23 September 1998 / Accepted: 25 September 1998
Keywords:  Frog skin epithelium  Fura 2  Short-circuit current  Na/Ca exchange  Cytosolic calcium
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