title: Imaging Nuclear Morphology and Organization in Cleared Plant Tissues Treated with Cell Cycle Inhibitors.
abstract: Synchronization of root cells through chemical treatment can generate a large number of cells blocked in specific cell cycle phases. In plants, this approach can be employed for cell suspension cultures and plant seedlings. To identify plant cells in the course of the cell cycle, especially during mitosis in meristematic tissues, chemical inhibitors can be used to block cell cycle progression. Herein, we present a simplified and easy-to-apply protocol to visualize mitotic figures, nuclei morphology, and organization in whole Arabidopsis root apexes. The procedure is based on tissue clearing, and fluorescent staining of nuclear DNA with DAPI. The protocol allows carrying out bulk analysis of nuclei and cell cycle phases in root cells and will be valuable to investigate mutants like overexpressing lines of genes disturbing the plant cell cycle.
authors.affiliation: Laboratório de Interação Molecular Planta-Praga, Embrapa Recursos Genéticos e Biotecnologia, PqEB, Av. W5 Norte final, Brasília, DF, 70770-900, Brazil.
authors.forename: José Dijair Antonino
authors.initials: JD
authors.lastname: de Souza Junior
authors.affiliation: Laboratório de Interação Molecular Planta-Praga, Embrapa Recursos Genéticos e Biotecnologia, PqEB, Av. W5 Norte final, Brasília, DF, 70770-900, Brazil.
authors.forename: Maria Fatima Grossi
authors.initials: MF
authors.lastname: de Sa
authors.affiliation: Institut National de la Recherche Agronomique, UMR 1355 ISA/Centre National de la Recherche Scientifique, UMR 7254 ISA/Université de Nice-Sophia Antipolis, UMR ISA, 400 route des Chappes, Sophia-Antipolis, France.
authors.forename: Gilbert
authors.initials: G
authors.lastname: Engler
authors.affiliation: Institut National de la Recherche Agronomique, UMR 1355 ISA/Centre National de la Recherche Scientifique, UMR 7254 ISA/Université de Nice-Sophia Antipolis, UMR ISA, 400 route des Chappes, Sophia-Antipolis, France. Janice.Almeida-Engler@sophia.inra.fr.|Institut Sophia Agrobiotech, 400 route des Chappes, BP 167, Sophia-Antipolis, Cedex, 0690, France. Janice.Almeida-Engler@sophia.inra.fr.
authors.forename: Janice de Almeida
authors.initials: Jde A
authors.lastname: Engler
mesh_terms: D017360:Arabidopsis / Q000166:cytology* / Q000187:drug effects / Q000648:ultrastructure*; D018929:Cell Culture Techniques / Q000379:methods; D002453:Cell Cycle / Q000187:drug effects; D002467:Cell Nucleus / Q000187:drug effects / Q000648:ultrastructure*; D018744:DNA, Plant / Q000032:analysis; D005456:Fluorescent Dyes / Q000032:analysis; D007211:Indoles / Q000032:analysis; D018613:Microscopy, Confocal / Q000379:methods*; D008938:Mitosis; D061848:Optical Imaging / Q000379:methods; D018517:Plant Roots / Q000166:cytology / Q000187:drug effects / Q000648:ultrastructure; D013194:Staining and Labeling / Q000379:methods
chemical_list: D018744:DNA, Plant; D005456:Fluorescent Dyes; D007211:Indoles; C007293:DAPI
keywords: Arabidopsis; Cell cycle; Inhibitors; Nuclei; Roots