Journal of Applied Physiology
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J Appl Physiol 92: 1603-1610, 2002; doi:10.1152/japplphysiol.00756.2001
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Vol. 92, Issue 4, 1603-1610, April 2002

Silver ions induce Ca2+ release from the SR in vitro by acting on the Ca2+ release channel and the Ca2+ pump

R. Tupling and H. Green

Department of Kinesiology, University of Waterloo, Waterloo, Ontario, Canada N2L 3G1

Silver nitrate (AgNO3) is a sulfhydryl oxidizing agent that induces a biphasic Ca2+ release from isolated sarcoplasmic reticulum (SR) vesicles by presumably oxidizing critical sulfhydryl groups in the Ca2+ release channel (CRC), causing the channel to open. To further examine the effects of AgNO3 on the CRC and the Ca2+-ATPase, Ca2+ release was measured in muscle homogenates prepared from rat hindlimb muscle using indo 1. Cyclopiazonic acid (CPA) and ruthenium red (RR) were used to inhibit the Ca2+-ATPase and block the CRC, respectively, before inducing Ca2+ release with both AgNO3 and 4-chloro-m-cresol (4-CMC), a releasing agent specific for the CRC. With AgNO3 and CPA, the early rapid rate of release (phase 1) was increased (P < 0.05) by 42% (314 ± 5 vs. 446 ± 39 µmol · g protein-1 · min-1), whereas the slower, more prolonged rate of release (phase 2) was decreased (P < 0.05) by 72% (267 ± 39 vs. 74 ± 7.7 µmol · g protein-1 · min-1). RR, in combination with AgNO3, had no effect on phase 1 (P > 0.05) (314 ± 51 vs. 334 ± 43 µmol · g protein-1 · min-1) and decreased phase 2 (P < 0.05) by 65% (245 ± 34 vs. 105 ± 8.2 µmol · g protein-1 · min-1). With 4-CMC, CPA had no effect (P > 0.05) on either phase 1 or 2. With addition of RR, phase 1 was reduced (P < 0.05) by 59% (2,468 ± 279 vs. 1,004 ± 87 µmol · g protein-1 · min-1), and RR completely blocked phase 2. Both AgNO3 and 4-CMC fully inhibited Ca2+-ATPase activity measured in homogenates. These findings indicate that AgNO3, but not 4-CMC, induces Ca2+ release by acting on both the CRC and the Ca2+-ATPase.

calcium ion cycling; calcium ion release; calcium ion uptake; silver nitrate


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