Gal80 intersectional regulation of cell-type specific expression in vertebrates |
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Authors: | Fujimoto Esther Gaynes Brooke Brimley Cameron J Chien Chi-Bin Bonkowsky Joshua L |
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Affiliation: | Division of Pediatric Neurology, Department of Pediatrics, University of Utah School of Medicine, Salt Lake City, Utah; Department of Neurobiology and Anatomy, University of Utah School of Medicine, Salt Lake City, Utah, USA. |
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Abstract: | 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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Keywords: | Gal80 zebrafish Gal4 Gal4‐VP16 intersectional expression conditional expression |
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