| Three-dimensional analysis of DNA replication foci: a comparative study on species and cell type in situ. | |
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MedLine Citation:
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PMID: 10817674 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Chromatin morphology of interphase nuclei in most cell lines of quail (Coturnix coturnix japonica) and chick (Gallus gallus domesticus) embryos shows typical interspecies differences. This intrinsic marker has been used in quail/chick chimerisation experiments, where also differences between cell types were noted. We asked whether similar differences between species and between cell types could be observed in S phase nuclei in situ. In this report, we used bromodeoxyuridine (BrdU) pulse labelling and anti-BrdU immunofluorescence to detect DNA replication foci in the nuclei of identified cells. In the central nervous system of 5- to 7-day-old quail and chick embryos, mesoderm-derived cells with strikingly different morphology and topographical distribution were studied: endothelial, i.e. polarised cells forming continuous tubes, and macrophages, i.e. non-polarised, ameboid or ramified individual cells. Using confocal microscopy, replication foci in the nuclei were assessed quantitatively and three-dimensional visualisations were produced. We consistently observed that: (1) chick, but never quail, nuclei displayed completely confluent replication sites, independent of cell type, and (2) macrophages, but not endothelial cells, had distinct perinucleolar replication sites, independent of species. We thus demonstrate a new relationship between cell type and spatial arrangement of DNA replication sites, and conclude that interspecies differences of chromatin distribution are conserved throughout S phase. Our results strongly recommend that work done on nuclear structure in vitro should not be extrapolated without reservation to cells in vivo. |
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Authors:
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P H Cossmann; P S Eggli; H Kurz |
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Publication Detail:
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Type: Comparative Study; Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: Histochemistry and cell biology Volume: 113 ISSN: 0948-6143 ISO Abbreviation: Histochem. Cell Biol. Publication Date: 2000 Mar |
Date Detail:
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Created Date: 2000-06-26 Completed Date: 2000-06-26 Revised Date: 2007-11-14 |
Medline Journal Info:
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Nlm Unique ID: 9506663 Medline TA: Histochem Cell Biol Country: GERMANY |
Other Details:
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Languages: eng Pagination: 195-205 Citation Subset: IM |
Affiliation:
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Anatomisches Institut, Lehrstuhl II, Albert-Ludwigs-Universität Freiburg, Germany. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Bromodeoxyuridine / metabolism Cell Nucleus / physiology Chick Embryo* / cytology Coturnix / embryology* DNA / analysis*, biosynthesis DNA Replication / physiology* Image Processing, Computer-Assisted Microscopy, Confocal S Phase / physiology* |
| Grant Support | |
ID/Acronym/Agency:
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N01-HD-2-3144/HD/NICHD NIH HHS |
| Chemical | |
Reg. No./Substance:
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59-14-3/Bromodeoxyuridine; 9007-49-2/DNA |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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