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Transgenic systems for unequivocal identification of cardiac myocyte nuclei and analysis of cardiomyocyte cell cycle status

Raulf, Alexandra; Horder, Hannes; Tarnawski, Laura LU ; Geisen, Caroline; Ottersbach, Annika; Roell, Wilhelm; Jovinge, Stefan LU ; Fleischmann, Bernd K. and Hesse, Michael (2015) In Basic Research in Cardiology 110(3). p.33-33
Abstract
Even though the mammalian heart has been investigated for many years, there are still uncertainties in the fields of cardiac cell biology and regeneration with regard to exact fractions of cardiomyocytes (CMs) at different developmental stages, their plasticity after cardiac lesion and also their basal turnover rate. A main shortcoming is the accurate identification of CM and the demonstration of CM division. Therefore, an in vivo model taking advantage of a live reporter-based identification of CM nuclei and their cell cycle status is needed. In this technical report, we describe the generation and characterization of embryonic stem cells and transgenic mice expressing a fusion protein of human histone 2B and the red fluorescence protein... (More)
Even though the mammalian heart has been investigated for many years, there are still uncertainties in the fields of cardiac cell biology and regeneration with regard to exact fractions of cardiomyocytes (CMs) at different developmental stages, their plasticity after cardiac lesion and also their basal turnover rate. A main shortcoming is the accurate identification of CM and the demonstration of CM division. Therefore, an in vivo model taking advantage of a live reporter-based identification of CM nuclei and their cell cycle status is needed. In this technical report, we describe the generation and characterization of embryonic stem cells and transgenic mice expressing a fusion protein of human histone 2B and the red fluorescence protein mCherry under control of the CM specific alpha MHC promoter. This fluorescence label allows unequivocal identification and quantitation of CM nuclei and nuclearity in isolated cells and native tissue slices. In ventricles of adults, we determined a fraction of <20 % CMs and binucleation of 77-90 %, while in atria a CM fraction of 30 % and a binucleation index of 14 % were found. We combined this transgenic system with the CAG-eGFP-anillin transgene, which identifies cell division and established a novel screening assay for cell cycle-modifying substances in isolated, postnatal CMs. Our transgenic live reporter-based system enables reliable identification of CM nuclei and determination of CM fractions and nuclearity in heart tissue. In combination with CAG-eGFP-anillin-mice, the cell cycle status of CMs can be monitored in detail enabling screening for proliferation-inducing substances in vitro and in vivo. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Genetically altered mice, Cardiac myocyte, Cell cycle, Proliferation, miRNA
in
Basic Research in Cardiology
volume
110
issue
3
pages
33 - 33
publisher
Springer
external identifiers
  • wos:000354607300013
  • scopus:84928738908
ISSN
0300-8428
DOI
10.1007/s00395-015-0489-2
language
English
LU publication?
yes
id
38e7a1e9-e099-4597-b9cf-19e4e1ff5596 (old id 7422066)
date added to LUP
2015-06-26 14:17:32
date last changed
2017-11-19 04:02:54
@article{38e7a1e9-e099-4597-b9cf-19e4e1ff5596,
  abstract     = {Even though the mammalian heart has been investigated for many years, there are still uncertainties in the fields of cardiac cell biology and regeneration with regard to exact fractions of cardiomyocytes (CMs) at different developmental stages, their plasticity after cardiac lesion and also their basal turnover rate. A main shortcoming is the accurate identification of CM and the demonstration of CM division. Therefore, an in vivo model taking advantage of a live reporter-based identification of CM nuclei and their cell cycle status is needed. In this technical report, we describe the generation and characterization of embryonic stem cells and transgenic mice expressing a fusion protein of human histone 2B and the red fluorescence protein mCherry under control of the CM specific alpha MHC promoter. This fluorescence label allows unequivocal identification and quantitation of CM nuclei and nuclearity in isolated cells and native tissue slices. In ventricles of adults, we determined a fraction of &lt;20 % CMs and binucleation of 77-90 %, while in atria a CM fraction of 30 % and a binucleation index of 14 % were found. We combined this transgenic system with the CAG-eGFP-anillin transgene, which identifies cell division and established a novel screening assay for cell cycle-modifying substances in isolated, postnatal CMs. Our transgenic live reporter-based system enables reliable identification of CM nuclei and determination of CM fractions and nuclearity in heart tissue. In combination with CAG-eGFP-anillin-mice, the cell cycle status of CMs can be monitored in detail enabling screening for proliferation-inducing substances in vitro and in vivo.},
  author       = {Raulf, Alexandra and Horder, Hannes and Tarnawski, Laura and Geisen, Caroline and Ottersbach, Annika and Roell, Wilhelm and Jovinge, Stefan and Fleischmann, Bernd K. and Hesse, Michael},
  issn         = {0300-8428},
  keyword      = {Genetically altered mice,Cardiac myocyte,Cell cycle,Proliferation,miRNA},
  language     = {eng},
  number       = {3},
  pages        = {33--33},
  publisher    = {Springer},
  series       = {Basic Research in Cardiology},
  title        = {Transgenic systems for unequivocal identification of cardiac myocyte nuclei and analysis of cardiomyocyte cell cycle status},
  url          = {http://dx.doi.org/10.1007/s00395-015-0489-2},
  volume       = {110},
  year         = {2015},
}