HgCl2-induced changes in cytosolic Ca2+ of cultured rabbit renal tubular cells

Fura 2 was used to measure changes in cytosolic Ca2+] (Ca2+]i) in cultured rabbit kidney proximal tubule cells exposed to HgCl2. Treatment with 2.5-10 microM HgCl2 resulted in an extracellular Ca2+] (Ca2+]e)-independent 2- to 12-fold increase in Ca2+]i above resting levels of about 100 nM. Treatment with 25-100 microM HgCl2 caused a rapid Ca2+]e-independent 10- to 12-fold increase in Ca2+]i within 1 min followed by a recovery to about 2-fold steady state by 3 min. With 25-100 microM HgCl2, both magnitude and rate of Ca2+ increase were similar, but recovery was greater with increasing doses. A slower, secondary increase in Ca2+]i followed which varied with HgCl2 concentration and required Ca2+]e. The first increase in Ca2+]i represents release from intracellular pools. Calcium channel blockers, calmodulin inhibitors, and mitochondrial inhibitors do not alter the patterns of Ca2+]i changes due to HgCl2. The recovery response with higher HgCl2 concentrations appears to be triggered by Hg2+ and not by the increased Ca2+]i. Sulfhydryl modifiers N-ethylmaleimide, PCMB and PCMBS produced Ca2+]e-independent Ca2+]i increases similar to those induced by low HgCl2 concentrations. Cell killing with HgCl2 was about 50% greater with normal Ca2+]e than with low Ca2+]e, suggesting that Ca2+]e influx is important in accelerating injury leading to cell death.