title: Live Imaging of Meiosis I in Late-Stage Drosophila melanogaster Oocytes.
abstract: Drosophila melanogaster has been studied for a century as a genetic model to understand recombination, chromosome segregation, and the basic rules of inheritance. However, it has only been about 25 years since the events that occur during nuclear envelope breakdown, spindle assembly, and chromosome orientation during D. melanogaster female meiosis I were first visualized by fixed cytological methods (Theurkauf and Hawley, J Cell Biol 116:1167-1180, 1992). Although these fixed cytological studies revealed many important details about the events that occur during meiosis I, they failed to elucidate the timing or order of these events. The development of protocols for live imaging of meiotic events within the oocyte has enabled collection of real-time information on the kinetics and dynamics of spindle assembly, as well as the behavior of chromosomes during prometaphase I. Here, we describe a method to visualize spindle assembly and chromosome movement during meiosis I by injecting fluorescent dyes to label microtubules and DNA into stage 12-14 oocytes. This method enables the events during Drosophila female meiosis I, such as spindle assembly and chromosome movement, to be observed in vivo, regardless of genetic background, with exceptional spatial and temporal resolution.
authors.affiliation: Stowers Institute for Medical Research, 1000 East 50th Street, Kansas City, MO, 64110, USA. sfh@stowers.org.
authors.forename: Stacie E
authors.initials: SE
authors.lastname: Hughes
authors.affiliation: Stowers Institute for Medical Research, 1000 East 50th Street, Kansas City, MO, 64110, USA.|Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, KS, 66160, USA.
authors.forename: R Scott
authors.initials: RS
authors.lastname: Hawley
mesh_terms: D000818:Animals; D059006:Chromosomes, Insect / Q000235:genetics / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000166:cytology* / Q000235:genetics; D005260:Female; D005456:Fluorescent Dyes / Q000378:metabolism; D008540:Meiosis; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods; D008870:Microtubules / Q000378:metabolism; D009865:Oocytes / Q000502:physiology*; D008941:Spindle Apparatus / Q000235:genetics / Q000378:metabolism
chemical_list: D005456:Fluorescent Dyes
keywords: Chromosome movement; Live imaging; Meiosis; Oocytes; Spindle assembly