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Functional fluorescent Ca2+ indicator proteins in transgenic mice under TET control
Authors: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
Institution:1 Max Planck Institute for Medical Research Heidelberg Germany;2 Universität Heidelberg Zentrum für Molekulare Biologie Heidelberg Germany;3 Department of Pharmacology and Howard Hughes Medical Institute, University of California San Diego, California United States of America;4 Brain Research Institute (RIKEN) Saitama Japan
Abstract: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.
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