Solhaug Anita, Gjessing Mona, Sandvik Morten, Eriksen Gunnar Sundstøl
Chemistry and Toxinology, Norwegian Veterinary Institute, P.O Box 64, 1431 Ås, Norway.
Aquatic Biosecurity, Norwegian Veterinary Institute, P.O Box 64, 1431 Ås, Norway.
Cytotechnology. 2023 Feb;75(1):63-75. doi: 10.1007/s10616-022-00560-0. Epub 2022 Nov 17.
In order to ensure the proper use and interpretation of results from laboratory test systems, it is important to know the characteristics of your test system. Here we compare mitochondria and the handling of reactive oxygen species (ROS) in two gill epithelial cell lines, the well-known RTgill-W1 cell line from Rainbow trout and the newly established ASG-10 cell line from Atlantic salmon. Rotenone was used to trigger ROS production. Rotenone reduced metabolic activity and induced cell death in both cell lines, with RTgill-W1 far more sensitive than ASG-10. In untreated cells, the mitochondria appear to be more fragmented in RTgill-W1 cells compared to ASG-10 cells. Furthermore, rotenone induced mitochondrial fragmentation, reduced mitochondria membrane potential (Δψm) and increased ROS generation in both cell lines. Glutathione (GSH) and catalase is important to maintain the cellular oxidative balance by eliminating hydrogen peroxide (HO). In response to rotenone, both GSH and catalase depletion were observed in the RTgill-W1 cells. In contrast, no changes were found in the GSH levels in ASG-10, while the catalase activity was increased. In summary, the two salmonid gill cell lines have different tolerance towards ROS, probably caused by differences in mitochondrial status as well as in GSH and catalase activities. This should be taken into consideration with the selection of experimental model and interpretation of results.
The online version contains supplementary material available at 10.1007/s10616-022-00560-0.
为确保实验室检测系统结果的正确使用和解读，了解检测系统的特性很重要。在此，我们比较了两种鳃上皮细胞系中线粒体及活性氧（ROS）的处理情况，这两种细胞系分别是虹鳟鱼中著名的RTgill-W1细胞系和新建立的大西洋鲑鱼ASG-10细胞系。鱼藤酮用于触发ROS产生。鱼藤酮降低了两种细胞系的代谢活性并诱导细胞死亡，其中RTgill-W1比ASG-10敏感得多。在未处理的细胞中，与ASG-10细胞相比，RTgill-W1细胞中的线粒体似乎更碎片化。此外，鱼藤酮在两种细胞系中均诱导了线粒体碎片化，降低了线粒体膜电位（Δψm）并增加了ROS生成。谷胱甘肽（GSH）和过氧化氢酶对于通过消除过氧化氢（HO）来维持细胞氧化平衡很重要。在鱼藤酮作用下，RTgill-W1细胞中观察到GSH和过氧化氢酶均耗竭。相比之下，ASG-10细胞中的GSH水平未发现变化，而过氧化氢酶活性增加。总之，这两种鲑科鱼类鳃细胞系对ROS的耐受性不同，这可能是由线粒体状态以及GSH和过氧化氢酶活性的差异所致。在选择实验模型和解读结果时应考虑到这一点。
在线版本包含可在10.1007/s10616-022-00560-0获取的补充材料。
