Calcium measurements in organelles with Ca2+-sensitive fluorescent proteins

被引:38
作者
Demaurex, N [1 ]
机构
[1] Univ Geneva, Dept Cell Physiol & Metab, CH-1211 Geneva, Switzerland
关键词
calcium; signalling; GFP; Ca2+ probes; endoplasmic reticulum;
D O I
10.1016/j.ceca.2005.06.026
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
The recent improvement in the design and use of genetically encoded fluorescent Ca2+ indicators should foster major progress in three aspects of Ca2+ signaling. At the subcellular level, ratiometric probes with expanded dynamics are now available to measure accurately the local Ca2+ changes occurring within specific cell compartments. These tools will allow to determine precisely the role of organelles and of cellular microdomains in Ca2+ handling. At the cellular level, the permanent labeling offered by the genetic probes enables large-scale, long-term Ca2+ measurements with robotic multiplexing technologies such as fluorescence plate readers or automated microscopes. This opens the way to large-scale pharmacological or genetic screens based on organelle-specific functional assays. At the whole animal level, probes with improved dynamics and reduced interference with endogenous proteins will allow to generate transgenic animals bearing Ca2+ sensitive indicators in specific cells and tissues. With this approach, Ca2+ signals can be recorded in neurons, heart, and pancreas of live animals during physiological and pathological stimulations. In this chapter, I will review the progress made in the design and use of genetic Ca2+ indicators, and discuss how these new tools provide an opportunity to challenge several unsolved questions in Ca2+ signaling, (C) 2005 Elsevier Ltd. All rights reserved.
引用
收藏
页码:213 / 222
页数:10
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