The effect of different kinds of electrolyte and non‐electrolyte solutions on the survival rate and morphology of zebrafish Danio rerio embryos

Abstract

The effect of electrolyte and non-electrolyte solutions on the survival and on the morphology of zebrafish Danio rerio embryos was investigated. Embryos in different ontogenetic stages were incubated in electrolyte (NaCl, KCl, MgCl2 and CaCl2) and non-electrolyte solutions [sucrose and polyvinylalcohol (PVA)] of different concentrations for 5-15 min. The embryos were hatched to the long-pec stage and the effective concentrations which caused a 50% decrease in embryo development (EC50) were determined. The morphometric changes, which were caused by the test solutions, were measured. Ion channel blockers were used to see if active ion transport played a role for embryo survival. Finally, dechorionated embryos were exposed to the test solutions to get indications about the importance of chorion and perivitelline space. For 12 hours post fertilization (hpf) embryos and a 15 min exposure period, EC50 was highest for MgCl2 (1.60 mol l(-1)), followed by sucrose (0.73 mol l(-1)), NaCl (0.49 mol l(-1)), KCl (0.44 mol l(-1)), CaCl2 (0.43 mol l(-1)) and PVA [0.0005 mol l(-1) (2.2%)]. EC50 were lower for early embryonic stages than for advanced stages for all solutions with exception of MgCl2 and sucrose. At the EC50, MgCl2 and CaCl2 solutions did not induce morphometric changes. NaCl and sucrose solutions induced reversible morphometric changes, which were compensated within 10 min. Only the EC50 of KCl and PVA solutions induced permanent morphometric changes, which could not be compensated. Incubation of embryos in electrolyte and non-electrolyte solutions together with ouabain (blocker of Na+- K+ ATPase), HgCl3 (dose-dependent inhibition of aquaporine channels), verapamil (inhibition of calcium and magnesium uptake) and amiloride (inhibition of sodium uptake) significantly decreased the per cent of embryos developing to the long-pec stage in comparison to the same solutions without blockers. Ouabain and HgCl(3) also induced morphometric changes. For dechorionated embryos the survival rates in water and in the different test solutions were similar to untreated embryos.