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Am J Physiol Cell Physiol 281: C972-C981, 2001;
0363-6143/01 $5.00
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Vol. 281, Issue 3, C972-C981, September 2001

Ca2+ regulation of gap junctional coupling in lens epithelial cells

Grant C. Churchill, Monica M. Lurtz, and Charles F. Louis

Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, Minnesota 55455

The quantitative effects of Ca2+ signaling on gap junctional coupling in lens epithelial cells have been determined using either the spread of Mn2+ that is imaged by its ability to quench the fluorescence of fura 2 or the spread of the fluorescent dye Alexa Fluor 594. Gap junctional coupling was unaffected by a mechanically stimulated cell-to-cell Ca2+ wave. Furthermore, when cytosolic Ca2+ concentration (Ca<UP><SUB>i</SUB><SUP>2+</SUP></UP>) increased after the addition of the agonist ATP, coupling was unaffected during the period that Ca<UP><SUB>i</SUB><SUP>2+</SUP></UP> was maximal. However, coupling decreased transiently ~5-10 min after agonist addition when Ca<UP><SUB>i</SUB><SUP>2+</SUP></UP> returned to resting levels, indicating that this transient decrease in coupling was unlikely due to a direct action of Ca<UP><SUB>i</SUB><SUP>2+</SUP></UP> on gap junctions. An increase in Ca<UP><SUB>i</SUB><SUP>2+</SUP></UP> mediated by the ionophore ionomycin that was sustained for several minutes resulted in a more rapid and sustained decrease in coupling (IC50 ~300 nM Ca2+, Hill coefficient of 4), indicating that an increase in Ca<UP><SUB>i</SUB><SUP>2+</SUP></UP> alone could regulate gap junctions. Thus Ca<UP><SUB>i</SUB><SUP>2+</SUP></UP> increases that occurred during agonist stimulation and cell-to-cell Ca2+ waves were too transient to mediate a sustained uncoupling of lens epithelial cells.

calcium; gap junction; manganese quench; fura 2


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