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Genetic mosaic with dual binary transcriptional systems in Drosophila

Nature Neuroscience volume 9, pages 703–709 (2006)Cite this article

Abstract

MARCM (mosaic analysis with a repressible cell marker) involves specific labeling of GAL80-minus and GAL4-positive homozygous cells in otherwise heterozygous tissues. Here we demonstrate how the concurrent use of two independent binary transcriptional systems may facilitate complex MARCM studies in the Drosophila nervous system. By fusing LexA with the VP16 acidic activation domain (VP16) or the GAL4 activation domain (GAD), we obtained both GAL80-insensitive and GAL80-suppressible transcriptional factors. LexA::VP16 can mediate MARCM-independent binary transgene induction in mosaic organisms. The incorporation of LexA::GAD into MARCM, which we call dual-expression-control MARCM, permits the induction of distinct transgenes in different patterns among GAL80-minus cells in mosaic tissues. Lineage analysis with dual-expression-control MARCM suggested the presence of neuroglioblasts in the developing optic lobes but did not indicate the production of glia by postembryonic mushroom body neuronal precursors. In addition, dual-expression-control MARCM with a ubiquitous LexA::GAD driver revealed many unidentified cells in the GAL4-GH146-positive projection neuron lineages.

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Figure 1: MARCM and binary transcriptional systems.
Figure 2: GAL4- versus LexA-based binary transcription systems.
Figure 3: Differential expression of distinct transgenes in ORNs versus PNs.
Figure 4: Labeling of GAL80-minus neurons and glia by dual-expression-control MARCM.
Figure 5: Dual-expression-control MARCM with tubP-LexA::GAD and GAL4-GH146.

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Acknowledgements

We thank J. Ma for pBTM116, S. Hanes for pSH18-34 and G. Struhl for GF51; L. Luo and L. Vosshall for sending us GAL4-GH146 and Or83b-GAL4 flies, respectively; C.T. Zugates and B. Leung for critical reading of the manuscript. This work was supported by the US National Institutes of Health and March of Dimes Birth Defects Foundation. T.L. is a Klingenstein Fellow.

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Authors and Affiliations

  1. Neuroscience Program and Department of Cell and Developmental Biology, University of Illinois at Urbana-Champaign, Urbana, 61801, IL, USA

    Sen-Lin Lai & Tzumin Lee

  2. Department of Neurobiology, University of Massachusetts Medical School, Worcester, 01605, Massachusetts, USA

    Sen-Lin Lai & Tzumin Lee

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  1. Sen-Lin Lai
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Correspondence to Tzumin Lee.

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Supplementary Fig. 1

Comparison of LexA::GAD/lexAop with GAL4/UAS (PDF 63 kb)

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Lai, SL., Lee, T. Genetic mosaic with dual binary transcriptional systems in Drosophila. Nat Neurosci 9, 703–709 (2006). https://doi.org/10.1038/nn1681

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