PMID- 25505384
OWN - NLM
STAT- MEDLINE
DCOM- 20150730
LR  - 20181113
IS  - 1662-5110 (Electronic)
IS  - 1662-5110 (Linking)
VI  - 8
DP  - 2014
TI  - Prolonged, brain-wide expression of nuclear-localized GCaMP3 for functional 
      circuit mapping.
PG  - 138
LID - 10.3389/fncir.2014.00138 [doi]
LID - 138
AB  - Larval zebrafish offer the potential for large-scale optical imaging of neural 
      activity throughout the central nervous system; however, several barriers 
      challenge their utility. First, ~panneuronal probe expression has to date only 
      been demonstrated at early larval stages up to 7 days post-fertilization (dpf), 
      precluding imaging at later time points when circuits are more mature. Second, 
      nuclear exclusion of genetically-encoded calcium indicators (GECIs) limits the 
      resolution of functional fluorescence signals collected during imaging. Here, we 
      report the creation of transgenic zebrafish strains exhibiting robust, nuclearly 
      targeted expression of GCaMP3 across the brain up to at least 14 dpf utilizing a 
      previously described optimized Gal4-UAS system. We confirmed both nuclear 
      targeting and functionality of the modified probe in vitro and measured its 
      kinetics in response to action potentials (APs). We then demonstrated in vivo 
      functionality of nuclear-localized GCaMP3 in transgenic zebrafish strains by 
      identifying eye position-sensitive fluorescence fluctuations in caudal hindbrain 
      neurons during spontaneous eye movements. Our methodological approach will 
      facilitate studies of larval zebrafish circuitry by both improving resolution of 
      functional Ca(2+) signals and by allowing brain-wide expression of improved 
      GECIs, or potentially any probe, further into development.
FAU - Kim, Christina K
AU  - Kim CK
AD  - Princeton Neuroscience Institute, Princeton University Princeton, NJ, USA ; 
      Department of Molecular Biology, Princeton University Princeton, NJ, USA.
FAU - Miri, Andrew
AU  - Miri A
AD  - Princeton Neuroscience Institute, Princeton University Princeton, NJ, USA ; 
      Department of Molecular Biology, Princeton University Princeton, NJ, USA.
FAU - Leung, Louis C
AU  - Leung LC
AD  - Department of Psychiatry and Behavioral Sciences, Stanford University Stanford, 
      CA, USA ; Center for Sleep Sciences, Stanford University Stanford, CA, USA.
FAU - Berndt, Andre
AU  - Berndt A
AD  - Department of Bioengineering, Stanford University Stanford, CA, USA.
FAU - Mourrain, Philippe
AU  - Mourrain P
AD  - Department of Psychiatry and Behavioral Sciences, Stanford University Stanford, 
      CA, USA ; Center for Sleep Sciences, Stanford University Stanford, CA, USA.
FAU - Tank, David W
AU  - Tank DW
AD  - Princeton Neuroscience Institute, Princeton University Princeton, NJ, USA ; 
      Department of Molecular Biology, Princeton University Princeton, NJ, USA.
FAU - Burdine, Rebecca D
AU  - Burdine RD
AD  - Department of Molecular Biology, Princeton University Princeton, NJ, USA.
LA  - eng
GR  - 5R90DA023419-04/DA/NIDA NIH HHS/United States
GR  - R01 DK090065/DK/NIDDK NIH HHS/United States
GR  - R01 MH060651/MH/NIMH NIH HHS/United States
GR  - R90 DA023419/DA/NIDA NIH HHS/United States
GR  - NS062798/NS/NINDS NIH HHS/United States
GR  - R01 NS062798/NS/NINDS NIH HHS/United States
GR  - R01 MH099647/MH/NIMH NIH HHS/United States
GR  - R01HD048584/HD/NICHD NIH HHS/United States
GR  - Howard Hughes Medical Institute/United States
GR  - R01 HD048584/HD/NICHD NIH HHS/United States
GR  - DK090065/DK/NIDDK NIH HHS/United States
GR  - R56 DK090065/DK/NIDDK NIH HHS/United States
GR  - MH099647/MH/NIMH NIH HHS/United States
GR  - R01MH060651/MH/NIMH NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20141126
PL  - Switzerland
TA  - Front Neural Circuits
JT  - Frontiers in neural circuits
JID - 101477940
RN  - 0 (Nerve Tissue Proteins)
RN  - 0 (Nuclear Proteins)
RN  - 0 (Zebrafish Proteins)
SB  - IM
MH  - Action Potentials/physiology
MH  - Animals
MH  - Animals, Genetically Modified
MH  - Brain/*physiology
MH  - Brain Mapping/*methods
MH  - Cell Nucleus/*metabolism
MH  - Cells, Cultured
MH  - Eye Movements/physiology
MH  - Fluorescence
MH  - HEK293 Cells
MH  - Humans
MH  - Nerve Tissue Proteins/genetics/*metabolism
MH  - Neural Pathways/physiology
MH  - Neurons/*physiology
MH  - Nuclear Proteins/genetics/*metabolism
MH  - Rats
MH  - Transfection
MH  - Zebrafish
MH  - Zebrafish Proteins/genetics/*metabolism
PMC - PMC4244806
OTO - NOTNLM
OT  - brain-wide expression
OT  - genetically encoded calcium indicators
OT  - in vivo calcium imaging
OT  - nuclear calcium signals
OT  - transgenic zebrafish
EDAT- 2014/12/17 06:00
MHDA- 2015/08/01 06:00
PMCR- 2014/01/01
CRDT- 2014/12/16 06:00
PHST- 2014/03/28 00:00 [received]
PHST- 2014/11/06 00:00 [accepted]
PHST- 2014/12/16 06:00 [entrez]
PHST- 2014/12/17 06:00 [pubmed]
PHST- 2015/08/01 06:00 [medline]
PHST- 2014/01/01 00:00 [pmc-release]
AID - 10.3389/fncir.2014.00138 [doi]
PST - epublish
SO  - Front Neural Circuits. 2014 Nov 26;8:138. doi: 10.3389/fncir.2014.00138. 
      eCollection 2014.