Rapid isolation of chloride cells from pinfish gill
Rapid isolation of chloride cells from pinfish gill
复制标题
从针鱼鳃中快速分离氯细胞
DOI：
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发表时间：
1978
期刊：
影响因子：
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通讯作者：
C. W. Philpott
中科院分区：
文献类型：
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作者：
S. Hootman;C. W. Philpott
从广盐性硬骨鱼Lagodon rhomboides鳃丝中获得了具有良好超微结构完整性的氯细胞富集组分。在不含Ca++、Mg++的平衡盐溶液中，通过温和的机械手段分离来自海水适应鱼的鳃上皮。通过Ficoll梯度对混合细胞悬浮液进行密度梯度离心，得到含有50 - 70%氯化物细胞的级分。该部分显示出钠加钾激活的腺苷三磷酸酶（Na+，K+-ATP酶）（一种在氯化物细胞中浓缩的酶）的3至4倍富集，高于可比的鳃匀浆值。从适应三分之一海水的鱼中分离氯细胞不太成功，因为这些细胞的尺寸较小且数量减少，尽管获得了至少2倍富集酶的组分。这些结果继续支持这样的观点：氯化细胞负责与硬骨鱼鳃上皮相关的渗透调节活性，并且这种重要功能是通过转运相关酶Na+，K+-ATP酶的活性介导的，其比活性随着渗透压的增加而增加。
Enriched fractions of chloride cells with good ultrastructural integrity have been obtained from gill filaments of the euryhaline teleost, Lagodon rhomboides. The branchial epithelium from seawater‐adapted fish was dissociated by gentle mechanical means in a Ca++, Mg++ ‐free balanced salt solution. Density gradient centrifugation of the mixed cell suspensions through a Ficoll gradient yielded a fraction containing between 50 and 70% chloride cells. This fraction showed a 3‐ to 4‐fold enrichment over comparable gill homogenate values for sodium plus potassium‐activated adenosinetriphosphatase, (Na+, K+‐ATPase), an enzyme concentrated in chloride cells. Isolation of chloride cells from fish adapted to one‐third seawater was less successful, due to the smaller size and reduced number of these cells, although fractions with at least a 2‐fold enrichment of the enzyme were obtained. These results continue to support the belief that chloride cells are responsible for the osmoregulatory activity associated with the branchial epithelium of teleosts and that this vital function is mediated through the activity of the transport associated enzyme, Na+, K+ ‐ATPase, the specific activity of which increases with osmotic stress.