Calcium transport in gill cells of Ucides cordatus, a mangrove crab living in variable salinity environments

Abstract

Crustaceans show discontinuous growth and have been used as a model system for studying cellular mechanisms of calcium transport, which is the main mineral found in their exoskeleton. Ucides cordatus, a mangrove crab, is naturally exposed to fluctuations in calcium and salinity. To study calcium transport in this species during isosmotic conditions, dissociated gill cells were marked with fluo-3 and intracellular Ca2+ change was followed by adding extracellular Ca2+ as CaCl2 (0, 0.1, 0.25, 0.50, 1.0 and 5 mM), together with different inhibitors. for control gill cells, Ca2+ transport followed Michaelis-Menten kinetics with V-max = 0.137 +/- 0.001 Delta Ca(2+)i (mu M x 22.10(4) cells(-1) x 180 s(-1); N = 4; r(2) = 0.99); K-m = 0.989 +/- 0.027 mM. the use of different inhibitors for gill cells showed that amiloride (Na+/Ca2+ exchange inhibitor) inhibited 80% of Ca2+ transport in gill cells (V-max). KB-R, an inhibitor of Ca influx in vertebrates, similarly caused a decrease in Ca2+ transport and verapamil (Ca2+ channel inhibitor) had no effect on Ca2+ transport, while nifedipine (another Ca2+ channel inhibitor) caused a 20% decrease in Ca2+ affinity compared to control values. Ouabain, on the other hand, caused no change in Ca2+ transport while vanadate increased the concentration of intracellular calcium through inhibition of Ca2+ efflux probably through the plasma membrane Ca2+-ATPase. Results show that transport kinetics for Ca2+ in these crabs under isosmotic conditions is lower compared to a hyper-regulator freshwater crab Dilocarcinus pagei studied earlier using fluorescent Ca2+ probes. These kinds of studies will help understanding the comparative mechanisms underlying the evolution of Ca transport in crabs living in different environments. (C) 2013 Elsevier Inc. All rights reserved.