RRC ID 40028
著者 Kim CK, Miri A, Leung LC, Berndt A, Mourrain P, Tank DW, Burdine RD.
タイトル Prolonged, brain-wide expression of nuclear-localized GCaMP3 for functional circuit mapping.
ジャーナル Front Neural Circuits
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 (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.
巻・号 8
ページ 138
公開日 2014-1-1
DOI 10.3389/fncir.2014.00138
PMID 25505384
PMC PMC4244806
MeSH Action Potentials / physiology Animals Animals, Genetically Modified Brain / physiology* Brain Mapping / methods* Cell Nucleus / metabolism* Cells, Cultured Eye Movements / physiology Fluorescence HEK293 Cells Humans Nerve Tissue Proteins / genetics Nerve Tissue Proteins / metabolism* Neural Pathways / physiology Neurons / physiology* Nuclear Proteins / genetics Nuclear Proteins / metabolism* Rats Transfection Zebrafish Zebrafish Proteins / genetics Zebrafish Proteins / metabolism*
IF 3.156
引用数 10
WOS 分野 NEUROSCIENCES
リソース情報
ゼブラフィッシュ Tg(huC:GVP)