Genetically Encoded Protein Sensors of Membrane Potential
- Genetically Encoded Protein Sensors of Membrane Potential
- Douglas Storace; 마소드; Zhou Han; Lei Jin; 래리코헨; Thom Hughes; 브래들리 베이커; Uhna Sung
- FP voltage sensors; Voltage-sensitive dyes; Genetically encoded
voltage sensors; Ciona; GFP; Membrane potential; Mammalian cells; Membrane expression; Thomas Knopfel; Ehud Isacoff
- Issue Date
- VOL 게제예, NO 게제예, 493-509
- Organic voltage-sensitive dyes offer very high spatial and temporal resolution for imaging neuronal function. However these dyes suffer from the drawbacks of non-specificity of cell staining and low accessibility of the dye to some cell types. Further progress in imaging activity is expected from the devel-opment of genetically encoded fluorescent sensors of membrane potential. Cell type specificity of expression of these fluorescent protein (FP) voltage sensors can be
obtained via several different mechanisms. One is cell type specificity of infection by individual virus subtypes. A second mechanism is specificity of promoter expression in individual cell types. A third, depends on the offspring of transgenic animals with cell type specific expression of cre recombinase mated with an animal that has the DNA for the FP voltage sensor in all of its cells but its expression is dependent on the recombinase activity. Challenges remain. First, the response time constants of many of the new FP voltage sensors are slower (2–10 ms) than those of organic dyes. This results in a relatively small fractional fluorescence change, F/F, for action potentials. Second, the largest signal presently available is only ~40% for a 100 mV depolarization and many of the new probes have signals that are substantially smaller. Large signals are especially important when attempting to detect fast events because the shorter measurement interval results in a relatively small number of detected photons and therefore a relatively large shot noise (see Chap. 1). Another kind of challenge has occurred when attempts were made to transition from one species to another or from one cell type to another or from cell culture to in vivo measurements. Several laboratories have recently described a number of novel FP voltage sensors. Here we attempt to critically review the current status of these develop-ments in terms of signal size, time course, a
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