Functional fluorescent Ca2+ indicator proteins in transgenic mice under TET control.
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.
Hasan
Mazahir T
MT
Max Planck Institute for Medical Research, Heidelberg, Germany. mhasan@mpimf-heidelberg.mpg.de
Friedrich
Rainer W
RW
Euler
Thomas
T
Larkum
Matthew E
ME
Giese
Günter
G
Both
Matthias
M
Duebel
Jens
J
Waters
Jack
J
Bujard
Hermann
H
Griesbeck
Oliver
O
Tsien
Roger Y
RY
Nagai
Takeharu
T
Miyawaki
Atsushi
A
Denk
Winfried
W
D000818:Animals; D001921:Brain / Q000378:metabolism*; D001931:Brain Mapping / Q000379:methods*; D002118:Calcium / Q000378:metabolism* / Q000502:physiology; D015220:Calcium Channels / Q000378:metabolism* / Q000502:physiology; D004318:Doxycycline; D036681:Fluorescence Recovery After Photobleaching; D015870:Gene Expression; D006367:HeLa Cells; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D008856:Microscopy, Fluorescence; D009830:Olfactory Bulb / Q000502:physiology*; D010775:Photic Stimulation; D012160:Retina / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology*; D014162:Transfection
D015220:Calcium Channels; D008164:Luminescent Proteins; D004318:Doxycycline; D002118:Calcium