title: Microscopy Methods for Analysis of Spindle Dynamics in Meiotic Drosophila Spermatocytes.
abstract: The spindle is a microtubule-based structure whose remodeling is required for partitioning the chromosomes and cytoplasm during meiosis. Characterizing microtubule behavior is fundamental to understanding how these tubulin polymers contribute to successful cell division. Here, a procedure is described for the imaging and analysis of spindle microtubule dynamics in cultures of living Drosophila melanogaster primary spermatocytes expressing tubulin tagged with enhanced green fluorescent protein. It employs time-lapse scanning confocal microscopy and the photobleaching of fiduciary marks onto fluorescently tagged microtubules. These labels are subsequently used to determine the sites and rates of kinetochore fiber growth and shrinkage during metaphase. This method can be readily applied to different microtubule populations, meiotic stages, and genetic backgrounds.
authors.affiliation: Institute of Fundamental Sciences, Massey University, Tennent Drive, Palmerston North, 4472, New Zealand. m.s.savoian@massey.ac.nz.
authors.forename: Matthew S
authors.initials: MS
authors.lastname: Savoian
mesh_terms: D000818:Animals; D004331:Drosophila melanogaster / Q000166:cytology* / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008297:Male; D008870:Microtubules / Q000378:metabolism; D013090:Spermatocytes / Q000166:cytology / Q000502:physiology*; D008941:Spindle Apparatus / Q000378:metabolism*; D059008:Time-Lapse Imaging / Q000295:instrumentation / Q000379:methods*
chemical_list: C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins
keywords: Fluorescent protein; Kinetochore fiber; Live cell imaging; Meiosis; Microtubule flux; Photobleaching; Time-lapse imaging