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

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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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References

  1. Brenner, S. Genetics 77, 71–94 (1974).

    CAS  PubMed  PubMed Central  Google Scholar 

  2. Jorgensen, E.M. & Mango, S.E. Nat. Rev. Genet. 3, 356–369 (2002).

    Article  CAS  PubMed  Google Scholar 

  3. Chalfie, M., Tu, Y., Euskirchen, G., Ward, W.W. & Prasher, D.C. Science 263, 802–805 (1994).

    Article  CAS  PubMed  Google Scholar 

  4. Pulak, R. Methods Mol. Biol. 351, 275–286 (2006).

    PubMed  Google Scholar 

  5. Abeliovich, A. & Hammond, R. Dev. Biol. 304, 447–454 (2007).

    Article  CAS  PubMed  Google Scholar 

  6. Nass, R. et al. J. Neurochem. 94, 774–785 (2005).

    Article  CAS  PubMed  Google Scholar 

  7. Nass, R., Hall, D.H., Miller, D.M., III & Blakely, R.D. Proc. Natl. Acad. Sci. USA 99, 3264–3269 (2002).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Chase, D.L., Pepper, J.S. & Koelle, M.R. Nat. Neurosci. 7, 1096–1103 (2004).

    Article  CAS  PubMed  Google Scholar 

  9. Davis, M.W. et al. BMC Genomics 6, 118 (2005).

    Article  PubMed  PubMed Central  Google Scholar 

  10. Zhao, C. & Emmons, S.W. Nature 373, 74–78 (1995).

    Article  CAS  PubMed  Google Scholar 

  11. Hack, M.A. et al. Nat. Neurosci. 8, 865–872 (2005).

    Article  CAS  PubMed  Google Scholar 

  12. Guenther, C. & Garriga, G. Development 122, 3509–3518 (1996).

    CAS  PubMed  Google Scholar 

  13. Chung, K., Crane, M.M. & Lu, H. Nat. Methods 5, 637–643 (2008).

    Article  CAS  PubMed  Google Scholar 

  14. Boulin, T. & Bessereau, J.L. Nat. Protoc. 2, 1276–1287 (2007).

    Article  CAS  PubMed  Google Scholar 

  15. Sarin, S., Prabhu, S., O'Meara, M.M., Pe'er, I. & Hobert, O. Nat. Methods advance online publication 1 August 2008 (doi:10.1038/nmeth.1249).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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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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