三疣梭子蟹na,k-atpase盐度调控机制的初步研究-preliminary study on the salinity control mechanism of na, k - atpase in portunus trituberculatus.docxVIP

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三疣梭子蟹na,k-atpase盐度调控机制的初步研究-preliminary study on the salinity control mechanism of na, k - atpase in portunus trituberculatus.docx

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三疣梭子蟹na,k-atpase盐度调控机制的初步研究-preliminary study on the salinity control mechanism of na, k - atpase in portunus trituberculatus

II II 1d 时，Na+,K+-ATPase 蛋白表达量并没有明显改变。由此可推断，Na+,K+-ATPase 酶对高盐胁迫更加敏感。盐度胁迫 1d 的鳃组织样本经过 H.E.染色，以 25 盐度的切片样本作为对照观察实验结果。结果表明：高盐度（40ppt）胁迫下的氯细胞数量明显增加；在低盐胁迫 1d 时，氯细胞则主要集中分布在鳃丝基部，细胞游离面向内凹陷。这暗示：Na+,K+-ATPase 酶对高盐胁迫更加敏感。综合来看，本项研究揭示：无论在高盐或低盐环境盐度胁迫下，三疣梭子蟹 Na+,K+-ATPase mRNA 水平呈现明显的增加；而仅在高盐胁迫初期，Na+,K+-ATPase 蛋白表达水平明显增加；而低盐胁迫初期，Na+,K+-ATPase 蛋白表达量并没有明显改变；Na+,K+-ATPase 酶对高盐胁迫更加敏感。另外，本项研究还揭示：高盐和低盐胁迫下，三疣梭子蟹 Na+,K+-ATPase 酶活的调节可能分为：被动应激期、主动调节期和适应期。三疣梭子蟹 Na+,K+-ATPase 参与了渗透调节过程。关键词三疣梭子蟹，Na+,K+-ATPase，盐度调控，半定量RT-PCR，酶活性，免疫组织化学 III III Preliminary Study of Portunus Trituberculatus Na+,K+- ATPase in Salinity Regulation Mechanism ABSTRACT Swimming crab, which has the great economic significance, is one of the most important marine aquaculture species in China. In the embryonic development and the whole larval stage, swimming crab is so sensitive to the change of environmental salinity that the slightest mistake may cause the delays in metamorphosis. High salinity can cause the difficulty of zoea truning into megalopa and the low megalopa metamorphosis rate. Therefore, for artificial breeding processes of swimming crab, salinity is a very important factor. Figuring out the mechanism of swimming crab adapted to environmental salinity has great significance in both theory and artificial breeding. This project studies the changes of Na+,K+-ATPase mRNA expression and Na+,K+- ATPase enzyme activities as well as the morphology and immunohistochemical characteristics under different salinity stress. The purpose of this study is to explore the role of swimming crab Na+,K+-ATPase in salinity regulation mechanism by gene level, protein level and organization level. After compared with control (25) salinity environment, the semi-quantitative RT- PCR results of Na+,K+-ATPase mRNA expression show that the increase in varying degrees of Na+,K+-ATPase mRNA expression level come from in both high and low salinity stress in the 6th gill, and the changes of expression level in muscle and intesti