title: Generation and Characterization of a Tissue-Specific Centrosome Indicator Mouse Line.
abstract: Centrosomes are major microtubule organizing centers (MTOCs) that play an important role in chromosome segregation during cell division. Centrosomes provide a stable anchor for microtubules, constituting the centers of the spindle poles in mitotic cells, and determining the orientation of cell division. However, visualization of centrosomes is challenging because of their small size. Especially in mouse tissues, it has been extremely challenging to observe centrosomes belonging to a specific cell type of interest among multiple comingled cell types. To overcome this obstacle, we generated a tissue-specific centrosome indicator. In this mouse line, a construct containing a floxed neomyocin resistance gene with a triplicate polyA sequence followed by an EGFP-Centrin1 fusion cassette was knocked into the Rosa locus. Upon Cre-mediated excision, EGFP-Centrin1 was expressed under the control of the Rosa locus. Experiments utilizing mouse embryo fibroblasts (MEFs) demonstrated the feasibility of real-time imaging, and showed that EGFP-Centrin1 expression mirrored the endogenous centrosome cycle, undergoing precisely one round of duplication through the cell cycle. Moreover, experiments using embryo and adult mouse tissues demonstrated that EGFP-Centrin1 specifically mirrors the localization of endogenous centrosomes. genesis 54:286-296, 2016. © 2016 The Authors. Genesis Published by Wiley Periodicals, Inc.
authors.affiliation: Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California, San Diego, La Jolla, California.
authors.forename: Maretoshi
authors.initials: M
authors.lastname: Hirai
authors.affiliation: Department of Medicine, University of California, San Diego, La Jolla, California.
authors.forename: Ju
authors.initials: J
authors.lastname: Chen
authors.affiliation: Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California, San Diego, La Jolla, California.|Department of Medicine, University of California, San Diego, La Jolla, California.|Department of Pharmacology, University of California, San Diego, La Jolla, California.
authors.forename: Sylvia M
authors.initials: SM
authors.lastname: Evans
mesh_terms: D000818:Animals; D002135:Calcium-Binding Proteins / Q000096:biosynthesis / Q000235:genetics*; D002455:Cell Division / Q000235:genetics*; D018385:Centrosome; D002868:Chromosomal Proteins, Non-Histone / Q000096:biosynthesis / Q000235:genetics*; D020090:Chromosome Segregation / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics; D051379:Mice; D022101:Microtubule-Organizing Center; D008938:Mitosis / Q000235:genetics*
chemical_list: D002135:Calcium-Binding Proteins; D002868:Chromosomal Proteins, Non-Histone; C485184:enhanced green fluorescent protein; C056608:caltractin; D049452:Green Fluorescent Proteins
keywords: cell cycle; cell division; centrin; centriole