Analyzing cell physiology in C. elegans with fluorescent ratiometric reporters.
Analyzing cell physiology in C. elegans with fluorescent ratiometric reporters.
复制标题
使用荧光比率报告仪分析线虫的细胞生理学。
DOI：
10.1016/j.ymeth.2014.05.012
复制
发表时间：
2014
期刊：
影响因子：
--
通讯作者：
Fisher,AlfredL
中科院分区：
文献类型：
--
作者：
Wang,Hongning;Karadge,Uma;Humphries4th,WilliamH;Fisher,AlfredL
Ratiometric fluorescent reporters have recently emerged a new technique to non-invasively measure aspects of cell physiology such as redox status, calcium levels, energy production, and NADH levels. These reporters consist of either a single or pair of fluorophores along with specific modifications, such as the addition of a protein domain which binds to a metabolite of interest, thereby producing gradual alterations in fluorescence in response to changes in the measured parameter. Measurement of the changes in fluorescence produces a quantitative read-out of the cellular environment. While these reporters were initially developed to easily visualize and track changes in cultured cells, several groups have adapted these reporters to use in Caenorhabditis elegans which opens a new avenue through which to explore cell physiology during development or aging, in response to changes in external environment, or in response to genetic manipulation. These reporters have the advantage of being easily targeted to any part of the worm, and becauseC. elegansis transparent both the reporters and changes in their fluorescence can be clearly observedin vivo. Here we discuss the application of ratiometric reporters toC. elegans, and outline a method to quantitatively measure changes in intracellular peroxide levels using the HyPer ratiometric reporter. However, these principles can be applied to alternate ratiometric reporters which are designed to measure either other chemical species or other cellular parameters.
