Hasan Mazahir T, Friedrich Rainer W, Euler Thomas, Larkum Matthew E, Giese Günter, Both Matthias, Duebel Jens, Waters Jack, Bujard Hermann, Griesbeck Oliver, Tsien Roger Y, Nagai Takeharu, Miyawaki Atsushi, Denk Winfried
Max Planck Institute for Medical Research, Heidelberg, Germany.
PLoS Biol. 2004 Jun;2(6):e163. doi: 10.1371/journal.pbio.0020163. Epub 2004 Jun 15.
Genetically encoded fluorescent calcium indicator proteins (FCIPs) are promising tools to study calcium dynamics in many activity-dependent molecular and cellular processes. Great hopes-for the measurement of population activity, in particular-have therefore been placed on calcium indicators derived from the green fluorescent protein and their expression in (selected) neuronal populations. Calcium transients can rise within milliseconds, making them suitable as reporters of fast neuronal activity. We here report the production of stable transgenic mouse lines with two different functional calcium indicators, inverse pericam and camgaroo-2, under the control of the tetracycline-inducible promoter. Using a variety of in vitro and in vivo assays, we find that stimuli known to increase intracellular calcium concentration (somatically triggered action potentials (APs) and synaptic and sensory stimulation) can cause substantial and rapid changes in FCIP fluorescence of inverse pericam and camgaroo-2.
基因编码的荧光钙指示剂蛋白(FCIPs)是研究许多活性依赖的分子和细胞过程中钙动力学的有前景的工具。因此,人们对群体活动的测量寄予厚望,尤其是对源自绿色荧光蛋白的钙指示剂及其在(选定的)神经元群体中的表达。钙瞬变可在数毫秒内上升,使其适合作为快速神经元活动的报告分子。我们在此报告了在四环素诱导型启动子控制下产生的具有两种不同功能钙指示剂(反向钙调蛋白和钙敏蛋白-2)的稳定转基因小鼠品系。通过各种体外和体内试验,我们发现已知能增加细胞内钙浓度的刺激(体细胞触发的动作电位(APs)以及突触和感觉刺激)可导致反向钙调蛋白和钙敏蛋白-2的FCIP荧光发生显著且快速的变化。