Dissection of Cell Division Processes in the One Cell Stage Caenorhabditis elegans Embryo by Mutational Analysis
| Autor: | Titus Kaletta, Pierre Gönczy, Ana Duran Amores, Tony Hyman, Heinke Schnabel, Ralf Schnabel |
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| Rok vydání: | 1999 |
| Předmět: |
Indirect
Embryo Nonmammalian Cell division Fluorescent Antibody Technique Video microscopy Biology Research Support P.H.S Models Biological Fluorescence microtubules video microscopy Models Pronuclear migration Animals genetics centrosomes Non-U.S. Gov't Caenorhabditis elegans Fluorescent Antibody Technique Indirect Anaphase mitosis Genetics Microscopy Nonmammalian Cell growth Caenorhabditis elegans/*embryology/genetics Chromosome Mapping Mutation Cell Biology Biological Cell biology Microscopy Fluorescence Embryo Centrosome Cell Division U.S. Gov't Centrosome separation Cytokinesis Regular Articles |
| Zdroj: | The Journal of Cell Biology ResearcherID |
| ISSN: | 1540-8140 0021-9525 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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