Targeting neural circuitry in zebrafish using GAL4 enhancer trapping

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Abstract

We present a pilot enhancer trap screen using GAL4 to drive expression of upstream activator sequence (UAS)-linked transgenes in expression patterns dictated by endogenous enhancers in zebrafish. The patterns presented include expression in small subsets of neurons throughout the larval brain, which in some cases persist into adult. Through targeted photoconversion of UAS-driven Kaede and variegated expression of UAS-driven GFP in single cells, we begin to characterize the cellular components of labeled circuits.

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Figure 1: Larval expression patterns for a selection of enhancer trap lines.
Figure 2: Analyses of trapped neural circuits.
Figure 3: Expression patterns are stable to adulthood.

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Acknowledgements

We thank A. Miyawaki for providing the Kaede open reading frame. N.C.C. thanks D. Stainier for support. This work was supported by a Sandler Opportunity Award and the 2006 Byers Award to H.B. and US National Institutes of Health National Research Service Award fellowship F32 EY015063 to E.K.S.

Author information

E.K.S. and H.B. conceived and designed the screen presented here and prepared this manuscript. E.K.S. performed subcloning of trapping constructs, injections, screening, establishment of transgenic lines, cloning of insertion sites and imaging. L.M. and A.A. participated in screening and L.M. established some transgenic lines. L.Z. performed dissection and sectioning of adult brains. N.G. and T.X. generated the UAS:Kaede and BGUG transgenic lines, respectively. K.A. and K.K. designed and generated the pT2KXIGΔ vector. N.C. performed subcloning of the 600bp hsp promoter.

Correspondence to Herwig Baier.

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Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Uniformity and consistency of expression. (PDF 525 kb)

Supplementary Table 1

Summary of ET lines. (PDF 19 kb)

Supplementary Table 2

Summary of expression patterns. (PDF 41 kb)

Supplementary Table 3

Integration sites. (PDF 20 kb)

Supplementary Methods (PDF 74 kb)

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Scott, E., Mason, L., Arrenberg, A. et al. Targeting neural circuitry in zebrafish using GAL4 enhancer trapping. Nat Methods 4, 323–326 (2007) doi:10.1038/nmeth1033

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