RRC ID 39979
Author Marquart GD, Tabor KM, Brown M, Strykowski JL, Varshney GK, LaFave MC, Mueller T, Burgess SM, Higashijima S, Burgess HA.
Title A 3D Searchable Database of Transgenic Zebrafish Gal4 and Cre Lines for Functional Neuroanatomy Studies.
Journal Front Neural Circuits
Abstract Transgenic methods enable the selective manipulation of neurons for functional mapping of neuronal circuits. Using confocal microscopy, we have imaged the cellular-level expression of 109 transgenic lines in live 6 day post fertilization larvae, including 80 Gal4 enhancer trap lines, 9 Cre enhancer trap lines and 20 transgenic lines that express fluorescent proteins in defined gene-specific patterns. Image stacks were acquired at single micron resolution, together with a broadly expressed neural marker, which we used to align enhancer trap reporter patterns into a common 3-dimensional reference space. To facilitate use of this resource, we have written software that enables searching for transgenic lines that label cells within a selectable 3-dimensional region of interest (ROI) or neuroanatomical area. This software also enables the intersectional expression of transgenes to be predicted, a feature which we validated by detecting cells with co-expression of Cre and Gal4. Many of the imaged enhancer trap lines show intrinsic brain-specific expression. However, to increase the utility of lines that also drive expression in non-neuronal tissue we have designed a novel UAS reporter, that suppresses expression in heart, muscle, and skin through the incorporation of microRNA binding sites in a synthetic 3' untranslated region. Finally, we mapped the site of transgene integration, thus providing molecular identification of the expression pattern for most lines. Cumulatively, this library of enhancer trap lines provides genetic access to 70% of the larval brain and is therefore a powerful and broadly accessible tool for the dissection of neural circuits in larval zebrafish.
Volume 9
Pages 78
Published 2015
DOI 10.3389/fncir.2015.00078
PMID 26635538
PMC PMC4656851
MeSH Animals Animals, Genetically Modified / anatomy & histology Animals, Genetically Modified / growth & development Animals, Genetically Modified / metabolism Brain / cytology Brain / metabolism DNA-Binding Proteins / genetics DNA-Binding Proteins / metabolism Databases, Factual* Gene Expression Imaging, Three-Dimensional / methods* MicroRNAs / genetics MicroRNAs / metabolism Microscopy, Confocal / methods Muscles / cytology Muscles / metabolism Myocardium / cytology Myocardium / metabolism Skin / cytology Skin / metabolism Software Transcription Factors / genetics Transcription Factors / metabolism Zebrafish / anatomy & histology* Zebrafish / growth & development Zebrafish / metabolism* Zebrafish Proteins / genetics Zebrafish Proteins / metabolism
IF 3.101
Times Cited 45
Zebrafish Tg(evx2-hs:Gal4) Tg(vglut2a:lRl-GFP)