Gal80 intersectional regulation of cell-type specific expression in vertebrates.

Gal80 intersectional regulation of cell-type specific expression in vertebrates.
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DOI:
10.1002/dvdy.22734
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发表时间:
2011-10
影响因子:
2.5
通讯作者:
Bonkowsky, Joshua L.
Bonkowsky, Joshua L.
中科院分区:
生物学3区
文献类型:
--
作者:
Fujimoto, Esther;Gaynes, Brooke;Brimley, Cameron J.;Chien, Chi-Bin;Bonkowsky, Joshua L.

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脊椎动物中枢神经系统(CNS)中特定神经元组的表征和功能操作仍然是理解复杂电路和功能的主要障碍。在斑马鱼中,Gal4/UAS 系统允许转基因和增强子捕获筛选的表达,但通常受到广泛表达域的限制。我们开发了一种利用 Gal80 抑制 Gal4 依赖性表达来进行细胞类型特异性表达的方法。我们表明,天然 Gal4 能够驱动强表达,Gal80 可以抑制这种表达,并且重叠的 Gal4 和 Gal80 表达可以在空间和遗传定义的神经元子集中实现“交叉”表达。我们还优化了 Gal80 在脊椎动物中的表达,用共表达的荧光标记追踪 Gal80 的表达,并使用 Gal80 的温度敏感等位基因暂时调节其功能。这些数据表明,Gal80 是对可用于绘制和操纵斑马鱼神经回路的遗传技术的有力补充。
Characterization and functional manipulation of specific groups of neurons in the vertebrate central nervous system (CNS) remains a major hurdle for understanding complex circuitry and functions. In zebrafish, the Gal4/UAS system has permitted expression of transgenes and enhancer trap screens, but is often limited by broad expression domains. We have developed a method for cell-type specific expression using Gal80 inhibition of Gal4-dependent expression. We show that native Gal4 is able to drive strong expression, that Gal80 can inhibit this expression, and that overlapping Gal4 and Gal80 expression can achieve “intersectional” expression in spatially and genetically defined subsets of neurons. We also optimize Gal80 for expression in vertebrates, track Gal80 expression with a co-expressed fluorescent marker, and use a temperature-sensitive allele of Gal80 to temporally regulate its function. These data demonstrate that Gal80 is a powerful addition to the genetic techniques available to map and manipulate neural circuits in zebrafish.
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