title: Visualization of the Nucleolus in Living Cells with Cell-Penetrating Fluorescent Peptides.
abstract: The nucleolus is the hallmark of nuclear compartmentalization and has been shown to exert multiple roles in cellular metabolism besides its main function as the place of ribosomal RNA synthesis and assembly of ribosomes. The nucleolus plays also a major role in nuclear organization as the largest compartment within the nucleus. The prominent structure of the nucleolus can be detected using contrast light microscopy providing an approximate localization of the nucleolus, but this approach does not allow to determine accurately the three-dimensional structure of the nucleolus in cells and tissues. Immunofluorescence staining with antibodies specific to nucleolar proteins albeit very useful is time consuming, normally antibodies recognize their epitopes only within a small range of species and is applicable only in fixed cells. Here, we present a simple method to selectively and accurately label this ubiquitous subnuclear compartment in living cells of a large range of species using a fluorescently labeled cell-penetrating peptide.
authors: Instituto de Medicina Molecular, Faculdade de Medicina, Universidade de Lisboa, 1649-028, Lisbon, Portugal.
authors: Robert M
authors: RM
authors: Martin
authors: Department of Biology, Technische Universität Darmstadt, Schnittspahnstr. 10, 64287, Darmstadt, Germany.
authors: Henry D
authors: HD
authors: Herce
authors: Department of Biology, Technische Universität Darmstadt, Schnittspahnstr. 10, 64287, Darmstadt, Germany.
authors: Anne K
authors: AK
authors: Ludwig
authors: Department of Biology, Technische Universität Darmstadt, Schnittspahnstr. 10, 64287, Darmstadt, Germany. cardoso@bio.tu-darmstadt.de.
authors: M Cristina
authors: MC
authors: Cardoso
mesh_terms: D000818:Animals; D015415:Biomarkers; D002460:Cell Line; D002466:Cell Nucleolus / Q000378:metabolism*; D057846:Cell-Penetrating Peptides / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D006801:Humans; D051379:Mice; D018613:Microscopy, Confocal; D057054:Molecular Imaging / Q000379:methods; D010455:Peptides / Q000378:metabolism; D012335:RNA, Ribosomal; D051381:Rats; D012270:Ribosomes; D013194:Staining and Labeling; D015027:Zebrafish
chemical_list: D015415:Biomarkers; D057846:Cell-Penetrating Peptides; D010455:Peptides; D012335:RNA, Ribosomal; C015462:polyarginine
keywords: Cell permeable nucleolar marker; Cell-penetrating peptide; Fluorescence microscopy; Living cells; Nucleolus; Poly-arginine