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© The Rockefeller University Press, 0021-9525/1999//927 $5.00
The Journal of Cell Biology, Volume 144, Number 5, , 1999 927-946

Dissection of Cell Division Processes in the One Cell Stage Caenorhabditis elegans Embryo by Mutational Analysis

Pierre Gönczy^*, Heinke Schnabel, Titus Kaletta, Ana Duran Amores^*, Tony Hyman^*,, and Ralf Schnabel

^* European Molecular Biology Laboratory, D-69117 Heidelberg, Germany; Max-Planck-Institut für Biochemie, D-82152 Martinsried, Germany; and Max-Planck-Institute for Cell Biology and Genetics, D-01307 Dresden, Germany

To identify novel components required for cell division processes in complex eukaryotes, we have undertaken an extensive mutational analysis in the one cell stage Caenorhabditis elegans embryo. The large size and optical properties of this cell permit observation of cell division processes with great detail in live specimens by simple differential interference contrast (DIC) microscopy. We have screened an extensive collection of maternal-effect embryonic lethal mutations on chromosome III with time-lapse DIC video microscopy. Using this assay, we have identified 48 mutations in 34 loci which are required for specific cell division processes in the one cell stage embryo. We show that mutations fall into distinct phenotypic classes which correspond, among others, to the processes of pronuclear migration, rotation of centrosomes and associated pronuclei, spindle assembly, chromosome segregation, anaphase spindle positioning, and cytokinesis. We have further analyzed pronuclear migration mutants by indirect immunofluorescence microscopy using antibodies against tubulin and ZYG-9, a centrosomal marker. This analysis revealed that two pronuclear migration loci are required for generating normal microtubule arrays and four for centrosome separation. All 34 loci have been mapped by deficiencies to distinct regions of chromosome III, thus paving the way for their rapid molecular characterization. Our work contributes to establishing the one cell stage C. elegans embryo as a powerful metazoan model system for dissecting cell division processes.

Key Words: genetics • video microscopy • mitosis • centrosomes • microtubules

Abbreviations used in this paper: DIC, differential interference contrast; eos, egg-osmotic sensitive; RNAi, RNA-mediated interference.

Some strains used in this work were obtained from the Caenorhabditis Genetics Center, which is funded by the National Institutes of Health National Center for Research Resources (NIH NCRR). Other strains were provided by the C. elegans Genetic Toolkit Project, which is funded by a grant from the NIH NCRR to Drs. Ann Rose, David L. Baillie, and Donald L. Riddle. Part of this project was supported by the Deutsche Forschungsgemeinschaft and the EMBL (European Molecular Biology Laboratory). Pierre Gönczy was a fellow from the European Molecular Biology Organization (ATLF 787-1995), the Human Frontier Science Program (LT-202/96), and the Swiss National Science Foundation (TMR 83EU-045376) during parts of this work.

Heinke Schnabel and Ralf Schnabel's present address is Technische Universität Braunschweig, Institut für Genetik, Spielmannstrasse 7, D-38106 Braunschweig, Germany. Titus Kaletta's present address is deVGen, Technologiepark 9, B-9052 Gent-Zwijnaarde, Belgium.

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