A cytological approach to studying meiotic recombination and chromosome dynamics in Arabidopsis thaliana male meiocytes in three dimensions

Aurélie Hurel, Dylan Phillips, Nathalie Vrielynck, Christine Mézard , Mathilde Grelon, Nicolas Christophorou

Research output: Contribution to journalArticlepeer-review

36 Citations (Scopus)
173 Downloads (Pure)

Abstract

During meiotic prophase I chromosomes undergo dramatic conformational changes that accompany chromosome condensation, pairing and recombination between homologs. These changes include the anchoring of telomeres to the nuclear envelope and their clustering to form a bouquet. In plants, these events have been studied and illustrated in intact meiocytes of species with large genomes. Arabidopsis thaliana is an excellent genetic model in which major molecular pathways that control synapsis and recombination between homologs have been uncovered. Yet the study of chromosome dynamics is hampered by current cytological methods that disrupt the three-dimensional (3D) architecture of the nucleus. Here we set up a protocol to preserve the 3D configuration of A. thaliana meiocytes. We showed that this technique is compatible with the use of a variety of antibodies that label structural and recombination proteins and were able to highlight the presence of clustered synapsis initiation centers at the nuclear periphery. By using fluorescence in situ hybridization we also studied the behavior of chromosomes during pre-meiotic G 2 and prophase I, revealing the existence of a telomere bouquet during A. thaliana male meiosis. In addition we showed that the number of telomeres in a bouquet and its volume vary greatly, thus revealing the complexity of telomere behavior during meiotic prophase I. Finally, by using probes that label subtelomeric regions of individual chromosomes, we revealed differential localization behaviors of chromosome ends. Our protocol opens new areas of research for investigating chromosome dynamics in A. thaliana meiocytes.

Original languageEnglish
Pages (from-to)385-396
Number of pages12
JournalPlant Journal
Volume95
Issue number2
Early online date22 Apr 2018
DOIs
Publication statusPublished - 31 Jul 2018

Keywords

  • 3D
  • Arabidopsis thaliana
  • Meiosis
  • chromosome dynamics
  • synapsis initiation centers
  • technical advance
  • telomere bouquet
  • Meiosis/genetics
  • Imaging, Three-Dimensional/methods
  • Arabidopsis/genetics
  • Prophase
  • Recombination, Genetic/genetics
  • Chromosomes, Plant/genetics
  • Telomere/metabolism

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