Influence of culture conditions on metal-induced responses in a cultured rainbow trout gill epithelium.
Influence of culture conditions on metal-induced responses in a cultured rainbow trout gill epithelium.
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培养条件对培养虹鳟鱼鳃上皮细胞金属诱导反应的影响。
DOI：
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发表时间：
2007
影响因子：
11.4
通讯作者：
C. Hogstrand
中科院分区：
文献类型：
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作者：
Paul A. Walker;N. Bury;C. Hogstrand
虹鳟鱼鳃细胞的原代培养技术进行了优化，以更好地代表完整的鳃在体内响应水性有毒金属。细胞接种密度和培养条件的修改导致鳃上皮细胞培养模型，显示经典的体内反应有毒金属。金属硫蛋白-A（MTA），金属硫蛋白-B（MTB），锌转运蛋白-1（ZnT-1），谷胱甘肽-S-转移酶（GST），葡萄糖-6-磷酸脱氢酶（G6 PD）的表达在mRNA水平上都表现出剂量依赖性的增加，以响应水性锌。在这些基因中，锌诱导的表达相对于对照的变化对于MTA、MTB和ZnT-1最大。MT表达也被银、铅、铜和镉诱导。细胞培养与淡水的顶侧保持净跨上皮内流的Ca 2+显示淡水鳟鱼鳃在体内，有一个积极的向内运动的Ca 2+。水性锌施加到顶端室减少净吸收的Ca 2+刺激流出组件。在发达的细胞培养系统中使用内源性金属响应基因表达和离子转运的抑制将促进金属鳃相互作用的研究，并可能被证明在天然沃茨的生物监测中具有未来的实际应用。
A primary culture technique for rainbow trout (Oncorhynchus mykiss) gill cells was optimized to better represent the intact gill in vivo in response to waterborne toxic metals. Modifications in cell seeding density and culture conditions resulted in a gill epithelial cell culture model, which displayed classic in vivo responses to toxic metals. Metallothionein-A (MTA), metallothionein-B (MTB), zinc transporter-1 (ZnT-1), glutathione-S-transferase (GST), and glucose-6-phosphate-dehydrogenase (G6PD) all showed dose-dependent increases in expression at the mRNA level in response to waterborne zinc. Of these genes, the change in zinc-induced expression relative to the control was greatest for MTA, MTB, and ZnT-1. MT expression was also induced by silver, lead, copper, and cadmium. Cells cultured with freshwater on the apical side maintained the net transepithelial influx of Ca2+ displayed by freshwater trout gills in vivo, and there was an active inward movement of Ca2+. Waterborne zinc applied to the apical compartment reduced the net uptake of Ca2+ by stimulating the efflux component. The use of endogenous metal-responsive gene expression and inhibition of ion transport in the developed cell culture system will facilitate studies of metal-gill interactions and may prove to have future practical applications within biomonitoring of natural waters.
DOI：
10.1152/ajplung.00267.2001
发表时间：
2002-02-01
影响因子：
4.9
作者：
St Croix, CM;Wasserloos, KJ;Pitt, BR
通讯作者：
Pitt, BR
影响因子：
5.8
作者：
Andrews, GK
通讯作者：
Andrews, GK