title: Using fast-acting temperature-sensitive mutants to study cell division in Caenorhabditis elegans.
abstract: Fast-acting temperature-sensitive (ts) mutations are powerful conditional tools for studying transient cellular processes such as cytokinesis. Fast-acting ts cytokinesis-defective mutants are functional at the permissive temperature; yet show a fully penetrant loss-of-function cytokinesis failure phenotype when upshifted to the restrictive temperature. Fast-acting ts mutations thus allow functional tunability and rapid and reversible protein inactivation by simply shifting the temperature at precise times throughout cell division. In this chapter, we describe several techniques and discuss various approaches for harnessing the power of fast-acting ts mutants to study cytokinesis in Caenorhabditis elegans using both simple passive heat transfer and more advanced fluidic-based thermal control systems. We also provide detailed protocols for standard dissection, mounting, and imaging of early worm embryos.
authors.affiliation: Columbia University, New York, NY, United States.
authors.forename: T
authors.initials: T
authors.lastname: Davies
authors.affiliation: Columbia University, New York, NY, United States.
authors.forename: S
authors.initials: S
authors.lastname: Sundaramoorthy
authors.affiliation: Columbia University, New York, NY, United States.
authors.forename: S N
authors.initials: SN
authors.lastname: Jordan
authors.affiliation: Columbia University, New York, NY, United States.
authors.forename: M
authors.initials: M
authors.lastname: Shirasu-Hiza
authors.affiliation: Institut Jacques Monod, Paris, France.
authors.forename: J
authors.initials: J
authors.lastname: Dumont
authors.affiliation: Columbia University, New York, NY, United States.
authors.forename: J C
authors.initials: JC
authors.lastname: Canman
mesh_terms: D000818:Animals; D017173:Caenorhabditis elegans / Q000235:genetics / Q000254:growth & development* / Q000648:ultrastructure; D048749:Cytokinesis / Q000235:genetics*; D004625:Embryo, Nonmammalian / Q000648:ultrastructure*; D057054:Molecular Imaging / Q000379:methods*; D009154:Mutation; D013696:Temperature
keywords: C. elegans; Cell division; CherryTemp; Cytokinesis; Permissive temperature; Rapid thermal upshift; Restrictive temperature; Temperature control device; Temperature-sensitive mutant; Therminator; Ts mutant