Sodium-calcium exchanger in cultured human retinal pigment epithelium

Experimental Eye Research
N J ManginiB G Kennedy

Abstract

Regulation of intracellular free Ca2+ concentration ([Ca2+]i) by an Na+/Ca2+ exchanger was studied in cultures of human retinal pigment epithelial cells using Ca(2+)-indicator dyes (fura-2 and fluo-3) and digital fluorescence imaging. Mean resting [Ca2+]i of cultured RPE in a control Ringer solution was 189 +/- 16 nM. Replacing extracellular Na+ with N-methyl-D-glucamine elicited a two-fold rise in [Ca2+]i; the magnitude of the [Na+]o-free-induced rise in [Ca2+]i varied as a function of extracellular [Ca2+]. The [Na+]o-free response was not significantly affected by the Ca2+ channel blocker nifedipine, or by pretreatment with thapsigargin which depletes intracellular Ca2+ stores. By contrast, the [Na+]o-free-induced rise in [Ca2+]i was significantly reduced by CBDMB, an amiloride derivative that is highly selective for Na+/Ca2+ exchange inhibition. These findings indicate that removal of extracellular Na+ promotes net [Ca2+]i gain via Na+/Ca2+ exchange. Western and Northern blot analyses, respectively, confirmed the presence of Na+/Ca2+ exchanger protein and mRNA in cultures of human RPE. Specifically, Western blot analysis of whole cell lysates of cultured RPE using a polyclonal antibody made against the canine cardiac exchang...Continue Reading

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Citations

Feb 14, 2006·Progress in Retinal and Eye Research·Geoffrey B Arden, Paul A Constable
Feb 3, 2000·Experimental Eye Research·B G KennedyN J Mangini
Aug 26, 2014·Experimental Eye Research·Nadine Reichhart, Olaf Strauss
Oct 8, 1999·Luminescence : the Journal of Biological and Chemical Luminescence·L J Kricka, P E Stanley
Jul 1, 2005·Physiological Reviews·Olaf Strauss
Aug 15, 2002·American Journal of Physiology. Cell Physiology·T X WengN K Wills
Feb 27, 2009·American Journal of Physiology. Cell Physiology·Mariko Sekiguchi-TonosakiJun Kosaka

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