Objective: To investigate how the sodium/calcium exchanger subtype 3 (NCX3) and its reverse mode contribute to the function of interstitial cells of Cajal (ICCs) from the rat bladder.
Methods: The study used 20 female Wistar rats. We observed the expression of the NCX3 expression in the bladder using reverse transcriptase-polymerase chain reaction and Western blotting. The NCX3 in ICCs was also confirmed by double-labeled fluorescence. NCX3 functions in reverse mode of ICCs were observed using confocal microscopy with preload fluo-3AM, and its currents were evaluated using the whole-cell patch clamp technique, with or without the NCX3 inhibitor KB-R7943 (5 and 30μM), with an afterward identification of ICCs using single-cell polymerase chain reaction.
Results: NCX3 was confirmed in rat bladder ICCs. The time required for the intracellular calcium concentration [Ca(2+)]i of NCX3 was enhanced by KB-R7943 (5μM, P ≤.01). Moreover, KB-R7943 (5 and 30μM) significantly decreased the currents generated by the reverse mode of NCX3 from the ICCs (P <.05).
Conclusion: NCX3 is expressed in rat bladder ICCs. The reverse mode of NCX3 can generate [Ca(2+)]i of the bladder ICCs.
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