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不同种类的电解质和非电解质溶液对斑马鱼 Danio rerio 胚胎存活率和形态的影响。

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

机构信息

Department for Organismic Biology, University of Salzburg, Hellbrunnerstrasse 34, 5020 Salzburg, Austria.

出版信息

J Fish Biol. 2009 Nov;75(7):1542-59. doi: 10.1111/j.1095-8649.2009.02354.x.

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.

摘要

研究了电解质和非电解质溶液对斑马鱼胚胎存活和形态的影响。将处于不同胚胎发生阶段的胚胎在不同浓度的电解质(NaCl、KCl、MgCl2 和 CaCl2)和非电解质(蔗糖和聚乙烯醇(PVA))溶液中孵育 5-15 分钟。将胚胎孵化到长鳍阶段,并确定导致胚胎发育减少 50%的有效浓度(EC50)。测量由测试溶液引起的形态变化。使用离子通道阻滞剂观察主动离子转运是否对胚胎存活起作用。最后,将去壳胚胎暴露于测试溶液中,以了解卵壳和卵周空间的重要性。对于受精后 12 小时(hpf)胚胎和 15 分钟暴露期,MgCl2 的 EC50 最高(1.60 mol l(-1)),其次是蔗糖(0.73 mol l(-1))、NaCl(0.49 mol l(-1))、KCl(0.44 mol l(-1))、CaCl2(0.43 mol l(-1))和 PVA[0.0005 mol l(-1)(2.2%)]。除了 MgCl2 和蔗糖外,所有溶液的早期胚胎阶段的 EC50 均低于晚期胚胎阶段。在 EC50 时,MgCl2 和 CaCl2 溶液不会引起形态变化。NaCl 和蔗糖溶液诱导可逆的形态变化,可在 10 分钟内得到补偿。只有 KCl 和 PVA 溶液的 EC50 诱导了永久性的形态变化,无法补偿。与不含阻滞剂的相同溶液相比,将胚胎与哇巴因(Na+-K+-ATP 酶阻滞剂)、HgCl3(水通道蛋白通道剂量依赖性抑制)、维拉帕米(钙和镁摄取抑制)和氨氯吡咪(钠摄取抑制)一起孵育在电解质和非电解质溶液中显著降低了胚胎发育到长鳍阶段的百分比。哇巴因和 HgCl(3)也诱导了形态变化。对于去壳的胚胎,在水中和不同测试溶液中的存活率与未处理的胚胎相似。

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