An easy protocol for studying chromatin and recombination protein dynamics during Arabidopsis thaliana meiosis: immunodetection of cohesins, histones and MLH1 - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue Cytogenetic and Genome Research Année : 2010

An easy protocol for studying chromatin and recombination protein dynamics during Arabidopsis thaliana meiosis: immunodetection of cohesins, histones and MLH1

Nathalie Vrielynck
  • Fonction : Auteur
  • PersonId : 1203077
Sylvia Le Guin
  • Fonction : Auteur
  • PersonId : 970571
Raphaël Mercier
Mathilde Grelon

Résumé

Plant meiosis studies have enjoyed a fantastic boom in recent years with the use of Arabidopsis thaliana as a model not only for molecular genetics and genomics but also for cytogenetics. In this article we describe a new protocol for immunolabelling meiotic proteins that allows the detection of a large range of proteins on strongly spread chromosomes throughout the entire meiotic process. We used this method to immunodetect MLH1, a crucial component of the meiotic recombination machinery, and found that it can be visualised as foci from pachytene to diakinesis, where it co-localises with chiasmata. The mean MLH1 foci number per meiotic cell at diakinesis was 8.4 for WS-4 and 9.95 for Col-0, with the number of foci per bivalent ranging from 1 to 5. We also analysed MLH1 distribution within bivalents and found that they were not restricted to specific chromosomal regions. The analysis of MLH1 foci formation in the Atzip4 mutant, where class I crossover (CO) formation is prevented, revealed that residual chiasmata were not labelled by MLH1, strongly suggesting that MLH1 antibodies only label class I COs in Arabidopsis. It thus appears that the 'obligatory CO' is systematically labeled by MLH1 and is generated through the class I pathway
Fichier non déposé

Dates et versions

hal-01203932 , version 1 (23-09-2015)

Identifiants

Citer

Liudmila Chelysheva, Laurie Grandont, Nathalie Vrielynck, Sylvia Le Guin, Raphaël Mercier, et al.. An easy protocol for studying chromatin and recombination protein dynamics during Arabidopsis thaliana meiosis: immunodetection of cohesins, histones and MLH1. Cytogenetic and Genome Research, 2010, 129 (1-3), pp.143-153. ⟨10.1159/000314096⟩. ⟨hal-01203932⟩
59 Consultations
0 Téléchargements

Altmetric

Partager

Gmail Facebook X LinkedIn More