PUBLICATION

Establishment of Gal4 transgenic zebrafish lines for analysis of development of cerebellar neural circuitry

Authors
Takeuchi, M., Matsuda, K., Yamaguchi, S., Asakawa, K., Miyasaka, N., Lal, P., Yoshihara, Y., Koga, A., Kawakami, K., Shimizu, T., Hibi, M.
ID
ZDB-PUB-141011-5
Date
2015
Source
Developmental Biology   397(1): 1-17 (Journal)
Registered Authors
Hibi, Masahiko, Kawakami, Koichi, Koga, Akihiko, Lal, Pradeep, Miyasaka, Nobuhiko, Shimizu, Takashi, Takeuchi, Miki, Yoshihara, Yoshihiro
Keywords
Cell migration, Cerebellum, Gal4, Neural circuits, Zebrafish
MeSH Terms
  • Purkinje Cells/cytology
  • Zebrafish/embryology
  • Zebrafish/genetics
  • DNA Transposable Elements
  • DNA-Binding Proteins/genetics*
  • Green Fluorescent Proteins/metabolism
  • Gene Expression Regulation, Developmental
  • Zebrafish Proteins/genetics*
  • Genes, Reporter
  • Neural Pathways
  • Transcription Factors/genetics*
  • Synapses
  • Animals, Genetically Modified
  • Genetic Techniques
  • Neurons/physiology
  • Cerebellum/embryology*
  • Cerebellum/physiology
  • Enhancer Elements, Genetic
  • Transgenes
  • Animals
  • Cell Movement
(all 21)
PubMed
25300581 Full text @ Dev. Biol.
Abstract
The cerebellum is involved in some forms of motor coordination and motor learning. Here we isolated transgenic (Tg) zebrafish lines that express a modified version of Gal4-VP16 (GFF) in the cerebellar neural circuits: granule, Purkinje, or eurydendroid cells, Bergmann glia, or the neurons in the inferior olive nuclei (IO) which send climbing fibers to Purkinje cells, with the transposon Tol2 system. By combining GFF lines with Tg lines carrying a reporter gene located downstream of Gal4 binding sequences (upstream activating sequence: UAS), we investigated the anatomy and developmental processes of the cerebellar neural circuitry. Combining an IO-specific Gal4 line with a UAS reporter line expressing the photoconvertible fluorescent protein Kaede demonstrated the contralateral projections of climbing fibers. Combining a granule cell-specific Gal4 line with a UAS reporter line expressing wheat germ agglutinin (WGA) confirmed direct and/or indirect connections of granule cells with Purkinje cells, eurydendroid cells, and IO neurons in zebrafish. Time-lapse analysis of a granule cell-specific Gal4 line revealed initial random movements and ventral migration of granule cell nuclei. Transgenesis of a reporter gene with another transposon Tol1 system visualized neuronal structure at a single cell resolution. Our findings indicate the usefulness of these zebrafish Gal4 Tg lines for studying the development and function of cerebellar neural circuits.
Genes / Markers
Figures
Figure Gallery (14 images) / 2
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Expression
Phenotype
No data available
Mutations / Transgenics
Allele Construct Type Affected Genomic Region
a9
    Complex
    nkgSA2AzGFF152BGtTransgenic Insertion
      nkgSAG6AGtTransgenic Insertion
        nkgSAIGFF23CGtTransgenic Insertion
          nkgsaizGFFM765BGtTransgenic Insertion
            nkhspGFF57AEtTransgenic Insertion
              nkhspGFFDMC28CEtTransgenic Insertion
                nkhspGFFDMC90AEtTransgenic Insertion
                  nkhspGFFDMC156AEtTransgenic Insertion
                    nkSAGFFLF128AEtTransgenic Insertion
                      1 - 10 of 22
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                      Human Disease / Model
                      No data available
                      Sequence Targeting Reagents
                      No data available
                      Fish
                      Antibodies
                      Name Type Antigen Genes Isotypes Host Organism
                      Ab1-neurodmonoclonalMouse
                      Ab1-pvalb7monoclonalMouse
                      Ab1-slc17a7polyclonalRabbit
                      Ab2-fabp7amonoclonal
                        Mouse
                        1 - 4 of 4
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                        Orthology
                        No data available
                        Engineered Foreign Genes
                        Marker Marker Type Name
                        EGFPEFGEGFP
                        GAL4FFEFGGAL4FF
                        GFPEFGGFP
                        KaedeEFGKaede
                        mCherryEFGmCherry
                        RFPEFGRFP
                        VenusEFGVenus
                        1 - 7 of 7
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                        Mapping
                        No data available