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Author Emiri, Andrew ♦ Tank, David W. ♦ Eleung, Louis ♦ Eberndt, Andre ♦ Emourrain, Philippe ♦ Burdine, Rebecca D. ♦ Kim, Christina Kay
Source Directory of Open Access Journals (DOAJ)
Content type Text
Publisher Frontiers Media S.A.
File Format HTM / HTML
Date Created 2015-09-11
Copyright Year ©2014
Language English
Subject Domain (in LCC) RC321-571
Subject Keyword Neuropsychiatry ♦ Biological psychiatry ♦ Neurosciences ♦ Transgenic zebrafish ♦ Internal medicine ♦ In vivo calcium imaging ♦ Medicine ♦ Nuclear calcium signals ♦ Genetically encoded calcium indicators ♦ Brain-wide expression
Abstract 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. 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 Ca2+ signals and by allowing brain-wide expression of improved GECIs, or potentially any probe, further into development.
ISSN 16625110
Age Range 18 to 22 years ♦ above 22 year
Educational Use Research
Education Level UG and PG ♦ Career/Technical Study
Learning Resource Type Article
Publisher Date 2014-11-01
e-ISSN 16625110
Journal Frontiers in Neural Circuits
Volume Number 8


Source: Directory of Open Access Journals (DOAJ)