Caffeine-induced Ca2+ oscillations in type I horizontal cells of the carp retina and the contribution of the store-operated Ca2+ entry pathway

Abstract

The mechanisms of release, depletion, and refilling of endoplasmic reticulum (ER) Ca2+ were investigated in type I horizontal cells of the carp retina using a fluo-3-based Ca2+ imaging technique. Exogenous application of caffeine, a ryanodine receptor agonist, induced oscillatory intracellular free Ca2+ concentration ([Ca 2+]i) responses in a duration- and concentration-dependent manner. In Ca2+-free Ringer's solution, [Ca2+] i transients could also be induced by a brief caffeine application, whereas subsequent caffeine application induced no [Ca2+]i increase, which implied that extracellular Ca2+ was required for ER refilling, confirming the necessity of a Ca2+ influx pathway for ER refilling. Depletion of ER Ca2+ by thapsigargin triggered a Ca 2+ influx which could be blocked by the store-operated channel inhibitor 2-APB, which proved the existence of the store-operated Ca 2+ entry pathway. Taken together, these results suggested that after being depleted by caffeine, the ER was replenished by Ca2+ influx via store-operated channels. These results reveal the fine modulation of ER Ca 2+ signaling, and the activation of the store-operated Ca 2+ entry pathway guarantees the replenishment of the ER so that the cell can be ready for response to the subsequent stimulus. © 2014 Lv et al.