RRC ID 33608
Author Takeuchi M, Matsuda K, Yamaguchi S, Asakawa K, Miyasaka N, Lal P, Yoshihara Y, Koga A, Kawakami K, Shimizu T, Hibi M.
Title Establishment of Gal4 transgenic zebrafish lines for analysis of development of cerebellar neural circuitry.
Journal 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.
Volume 397(1)
Pages 1-17
Published 2015-1-1
DOI 10.1016/j.ydbio.2014.09.030
PII S0012-1606(14)00499-0
PMID 25300581
MeSH Animals Animals, Genetically Modified Cell Movement Cerebellum / embryology* Cerebellum / physiology DNA Transposable Elements DNA-Binding Proteins / genetics* Enhancer Elements, Genetic Gene Expression Regulation, Developmental Genes, Reporter Genetic Techniques Green Fluorescent Proteins / metabolism Neural Pathways Neurons / physiology Purkinje Cells / cytology Synapses Transcription Factors / genetics* Transgenes Zebrafish / embryology Zebrafish / genetics Zebrafish Proteins / genetics*
IF 2.896
Times Cited 32
Zebrafish rw0318(Tg(UAS:AcGFP-P2A-WGA)) gSAG6A SAGFF(LF)128A hspGFF57A SAGFF(LF)157B hspGFFDMC90A hspzGFFgDMC156A hspGFFDMC28C SAGFF(LF)251A SAGFF(LF)226B nub6Tg(Tg(alloca:GFF)) rk22Tg(Tg(aldoca:gap43-Venus)) nub5Tg(Tg(aldoca:gap43-mCherry))