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Automated screening for mutants affecting dopaminergic-neuron specification in C. elegans

Nature Methods volume 5, pages 869–872 (2008)Cite this article

Abstract

We describe an automated method to isolate mutant Caenorhabditis elegans that do not appropriately execute cellular differentiation programs. We used a fluorescence-activated sorting mechanism implemented in the COPAS Biosort machine to isolate mutants with subtle alterations in the cellular specificity of GFP expression. This methodology is considerably more efficient than comparable manual screens and enabled us to isolate mutants in which dopamine neurons do not differentiate appropriately.

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Figure 1: Screening for dopaminergic cell-fate mutants.
Figure 2: Relative fluorescence intensity plots between red and green channels of sorted worms.
Figure 3: Phenotypes of isolated dopy mutants.

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Acknowledgements

We thank A. Kruyer, A. Pretorian, J. Recio and Q. Chen for technical assistance, S. Sarin for calculating the degree of saturation, V. Bertrand and F. Zhang for communicating unpublished results, and S. Mitani (Tokyo Women's Medical University School of Medicine) for providing lin-32 deletion alleles. This work was funded by the US National Institutes of Health (R01NS039996-05; R01NS050266-03), the Howard Hughes Medical Institute, an European Molecular Biology Organization long-term fellowship to M.D. and N.F., and a Marie Curie Outgoing International fellowship to N.F.

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biophysics, Howard Hughes Medical Institute, Columbia University Medical Center, 701 W. 168th Street, New York, 10032, New York, USA

    Maria Doitsidou, Nuria Flames, Albert C Lee, Alexander Boyanov & Oliver Hobert

Authors
  1. Maria Doitsidou
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  2. Nuria Flames
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  3. Albert C Lee
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  4. Alexander Boyanov
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  5. Oliver Hobert
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Contributions

M.D. conducted the genetic screens (manual and sorter), implemented the sorter strategy, mapped and characterized mutants, and co-wrote the paper; N.F. implemented the sorter strategy and conducted sorter screens; A.C.L. conducted manual screens; A.B. assisted in mapping mutants; and O.H. initiated and supervised the project, and co-wrote the paper.

Corresponding authors

Correspondence to Maria Doitsidou or Oliver Hobert.

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Supplementary Text and Figures

Supplementary Figures 1–2, Supplementary Tables 1–6, Supplementary Methods (PDF 3378 kb)

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Doitsidou, M., Flames, N., Lee, A. et al. Automated screening for mutants affecting dopaminergic-neuron specification in C. elegans. Nat Methods 5, 869–872 (2008). https://doi.org/10.1038/nmeth.1250

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