Establishment of Gal4 transgenic zebrafish lines for analysis of development of cerebellar neural circuitry
Establishment of Gal4 transgenic zebrafish lines for analysis of development of cerebellar neural circuitry
复制标题
用于分析小脑神经回路发育的 Gal4 转基因斑马鱼系的建立
DOI：
10.1016/j.ydbio.2014.09.030
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发表时间：
2015
影响因子：
2.7
通讯作者：
M.
中科院分区：
文献类型：
--
作者：
Takeuchi;M.;Matsuda;K.;Yamaguchi;S.;Asakawa;K.;Miyasaka;N.;Lal;P.;Yoshihara;Y.;Koga;A.;Kawakami;K.;Shimizu;T.;and Hibi;M.
小脑参与某些形式的运动协调和运动学习。在这里，我们分离了转基因(TG)斑马鱼，这些斑马鱼在小脑神经回路中表达Gal4-VP16(GFF)的修饰版本：颗粒、浦肯野或树突状细胞，Bergmann胶质细胞，或转座子Tol2系统，下橄榄核(IO)中的神经元向蒲肯野细胞发送攀升纤维。通过将GFF系与携带Gal4结合序列下游报告基因(上游激活序列：UAS)的TG系相结合，我们研究了小脑神经回路的解剖和发育过程。结合IO特异的Gal4系和表达光可转换荧光蛋白Kaeed的UAS报告系，展示了攀爬纤维的对侧投射。将颗粒细胞特异的Gal4细胞系与表达小麦胚凝集素(WGA)的UAS报告系相结合，证实了斑马鱼颗粒细胞与蒲肯野细胞、树突状细胞和IO神经元的直接和/或间接联系。对颗粒细胞特异性Gal4细胞系的时间推移分析表明，颗粒细胞核最初的随机运动和腹向迁移。一个报告基因与另一个转座子Tol1系统的转基因在单细胞分辨率下可视化了神经元结构。我们的发现表明这些斑马鱼Gal4Tg线对于研究小脑神经回路的发育和功能是有用的。
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.