Open Access Highly Accessed Research article

Large-scale mapping of mutations affecting zebrafish development

Robert Geisler1*, Gerd-Jörg Rauch12, Silke Geiger-Rudolph1, Andrea Albrecht13, Frauke van Bebber14, Andrea Berger1, Elisabeth Busch-Nentwich15, Ralf Dahm16, Marcus PS Dekens17, Christopher Dooley1, Alexandra F Elli18, Ines Gehring1, Horst Geiger1, Maria Geisler1, Stefanie Glaser1, Scott Holley19, Matthias Huber110, Andy Kerr1, Anette Kirn111, Martina Knirsch112, Martina Konantz1, Axel M Küchler113, Florian Maderspacher114, Stephan C Neuhauss115, Teresa Nicolson116, Elke A Ober117, Elke Praeg118, Russell Ray119, Brit Rentzsch120, Jens M Rick121, Eva Rief1, Heike E Schauerte122, Carsten P Schepp123, Ulrike Schönberger1, Helia B Schonthaler124, Christoph Seiler125, Samuel Sidi126, Christian Söllner127, Anja Wehner128, Christian Weiler1 and Christiane Nüsslein-Volhard1

Author Affiliations

1 Department 3 – Genetics, Max-Planck-Institut für Entwicklungsbiologie, Spemannstr. 35/III, 72076 Tübingen, Germany

2 Department of Internal Medicine III – Cardiology, University of Heidelberg, Im Neuenheimer Feld 350, 69120 Heidelberg, Germany

3 Max Planck Institute for Molecular Genetics, Ihnestr. 63-73, 14195 Berlin, Germany

4 Laboratory for Alzheimer's and Parkinson's Disease Research, Adolf-Butenandt-Institute, Department of Biochemistry, LMU, Schillerstr. 44, 80336 München, Germany

5 Team 31 – Vertebrate Development and Genetics, Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus Hinxton, Cambridge, CB10 1SA, UK

6 Center for Brain Research – Division of Neuronal Cell Biology, Medical University of Vienna, Spitalgasse 4, 1090 Vienna, Austria

7 Centre for Cellular and Molecular Dynamics, Department of Anatomy and Developmental Biology, University College London, Gower St., London WC1E 6BT, UK

8 3. Physikalisches Institut, Universität Stuttgart, Pfaffenwaldring 57, 70569 Stuttgart, Germany

9 Department of Molecular, Cellular and Developmental Biology, Yale University, P.O. Box 208103, New Haven, CT 06520-8103, USA

10 Institut für Klinische Pharmakologie und Toxikologie, Charité – Universitätsmedizin Berlin, Campus Benjamin Franklin, Hindenburgdamm 30, 12200 Berlin, Germany

11 NMI – Natural and Medical Science Institute at the University of Tübingen, Markwiesenstr. 55, 72770 Reutlingen, Germany

12 Institute of Physiology Dept. II and Tübingen Hearing Research Centre THRC, University of Tübingen, Elfriede-Aulhorn-Str. 5, 72076 Tübingen, Germany

13 Institute of Pathology, Rikshospitalet, Sognsvannveien 20, 0027 Oslo, Norway

14 Current Biology, Elsevier London, 84 Theobald's Rd., London WC1X 8RR, UK

15 Institute of Zoology, University of Zurich, Winterthurerstr. 190, 8057 Zürich, Switzerland

16 Oregon Hearing Research Center and Vollum Institute, Oregon Health & Science University, 3181 SW Sam Jackson Pk. Rd., Portland, OR 97239, USA

17 Division of Developmental Biology, National Institute for Medical Research, The Ridgeway, Mill Hill, London NW7 1AA, UK

18 Laboratory for Magnetic Brain Stimulation, Behavioral Neurology Unit, Beth Israel Deaconess Medical Center, Harvard Medical School, 330 Brookline Ave., Boston, MA 02215, USA

19 Howard Hughes Medical Institute, University of Utah, 15 North 2030 East, Salt Lake City, UT 84112, USA

20 MDC – Max-Delbrück-Centrum für Molekulare Medizin, Berlin-Buch, Robert-Rössle-Str. 10, 13092 Berlin, Germany

21 Cellzome AG, Meyerhofstr. 1, 69117 Heidelberg, Germany

22 Ingenium Pharmaceuticals AG, Fraunhoferstr. 13, 82152 Martinsried, Germany

23 Abt. Kinderheilkunde I, Children's Hospital, University of Tübingen, Hoppe-Seyler-Str. 1, 72076 Tübingen, Germany

24 IMP – Research Institute of Molecular Pathology, Dr. Bohr-Gasse 7, 1030 Vienna, Austria

25 Department of Medicine, University of Pennsylvania School of Medicine, 1230 Biomedical Research Building II/III, 421 Curie Blvd., Philadelphia, PA 19104, USA

26 Department of Pediatric Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Mayer Building 630, 44 Binney St., Boston, MA 02115, USA

27 Team 30 – Vertebrate functional proteomics laboratory, Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus Hinxton, Cambridge, CB10 1SA, UK

28 Department of Cell Biology, Max-Planck-Institute of Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany

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BMC Genomics 2007, 8:11  doi:10.1186/1471-2164-8-11

Published: 9 January 2007

Abstract

Background

Large-scale mutagenesis screens in the zebrafish employing the mutagen ENU have isolated several hundred mutant loci that represent putative developmental control genes. In order to realize the potential of such screens, systematic genetic mapping of the mutations is necessary. Here we report on a large-scale effort to map the mutations generated in mutagenesis screening at the Max Planck Institute for Developmental Biology by genome scanning with microsatellite markers.

Results

We have selected a set of microsatellite markers and developed methods and scoring criteria suitable for efficient, high-throughput genome scanning. We have used these methods to successfully obtain a rough map position for 319 mutant loci from the Tübingen I mutagenesis screen and subsequent screening of the mutant collection. For 277 of these the corresponding gene is not yet identified. Mapping was successful for 80 % of the tested loci. By comparing 21 mutation and gene positions of cloned mutations we have validated the correctness of our linkage group assignments and estimated the standard error of our map positions to be approximately 6 cM.

Conclusion

By obtaining rough map positions for over 300 zebrafish loci with developmental phenotypes, we have generated a dataset that will be useful not only for cloning of the affected genes, but also to suggest allelism of mutations with similar phenotypes that will be identified in future screens. Furthermore this work validates the usefulness of our methodology for rapid, systematic and inexpensive microsatellite mapping of zebrafish mutations.