title: Measurement and 3D-visualization of cell-cycle length using double labelling with two thymidine analogues applied in early heart development.
abstract: Organ development is a complex spatial process in which local differences in cell proliferation rate play a key role. Understanding this role requires the measurement of the length of the cell cycle at every position of the three-dimensional (3D) structure. This measurement can be accomplished by exposing the developing embryo to two different thymidine analogues for two different durations immediately followed by tissue fixation. This paper presents a method and a dedicated computer program to measure the resulting labelling indices and subsequently calculate and visualize local cell cycle lengths within the 3D morphological context of a developing organ. By applying this method to the developing heart, we show a large difference in cell cycle lengths between the early heart tube and the adjacent mesenchyme of the pericardial wall. Later in development, a local increase in cell size was found to be associated with a decrease in cell cycle length in the region where the chamber myocardium starts to develop. The combined application of halogenated-thymidine double exposure and image processing enables the automated study of local cell cycle parameters in single specimens in a full 3D context. It can be applied in a wide range of research fields ranging from embryonic development to tissue regeneration and cancer research.
authors.affiliation: Department of Anatomy, Embryology & Physiology, Academic Medical Center, Amsterdam, The Netherlands.
authors.forename: Bouke A
authors.initials: BA
authors.lastname: de Boer
authors.forename: Gert
authors.initials: G
authors.lastname: van den Berg
authors.forename: Alexandre T
authors.initials: AT
authors.lastname: Soufan
authors.forename: Piet A J
authors.initials: PA
authors.lastname: de Boer
authors.forename: Jaco
authors.initials: J
authors.lastname: Hagoort
authors.forename: Maurice J B
authors.initials: MJ
authors.lastname: van den Hoff
authors.forename: Antoon F M
authors.initials: AF
authors.lastname: Moorman
authors.forename: Jan M
authors.initials: JM
authors.lastname: Ruijter
mesh_terms: D000818:Animals; D002453:Cell Cycle; D002642:Chick Embryo; D003198:Computer Simulation; D006321:Heart / Q000196:embryology*; D021621:Imaging, Three-Dimensional; D057054:Molecular Imaging; D009206:Myocardium / Q000378:metabolism*; D038081:Organogenesis / Q000502:physiology; D013194:Staining and Labeling; D013936:Thymidine / Q000031:analogs & derivatives
chemical_list: D013936:Thymidine