Imaging and Quantitation of Assembly Dynamics of the Centromeric Histone H3 Variant CENP-A in Drosophila melanogaster Spermatocytes by Immunofluorescence and Fluorescence In-Situ Hybridization (Immuno-FISH)

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Abstract

This chapter describes a method used to assay the cell cycle dynamics of the centromeric histone H3 variant CENP-A in meiosis using Drosophila males as the experimental system. Specifically, we describe a method that combines Immunofluorescence (IF) and Fluorescence in-situ Hybridization (FISH) protocols, performed on fixed Drosophila testes. An advantage of this protocol is the ability to localize individual centromeres on the four Drosophila homologous chromosomes that form distinct nuclear territories in spermatocytes. We also describe a method to quantify CENP-A focal intensities using Image J software.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages327-337
Number of pages11
DOIs
Publication statusPublished - 2018

Publication series

NameMethods in Molecular Biology
Volume1832
ISSN (Print)1064-3745

Keywords

  • CENP-A
  • Centromere
  • Chromosome segregation
  • Drosophila melanogaster
  • Meiosis
  • Prophase I
  • Spermatogenesis

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