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Measurement of S-phase duration of adult stem cells in the flatworm Macrostomum lignano by double replication labelling and quantitative colocalization analysis

Freija Verdoodt (UGent) , Maxime Willems (UGent) , Ineke Dhondt (UGent) , Wouter Houthoofd (UGent) , Wim Bert (UGent) and Winnok De Vos (UGent)
(2012) CELL BIOLOGY INTERNATIONAL. 36(12). p.1251-1259
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Abstract
Platyhelminthes are highly attractive models for addressing fundamental aspects of stem cell biology in vivo. These organisms possess a unique stem cell system comprised of neoblasts that are the only proliferating cells during adulthood. We have investigated T-s (S-phase duration) of neoblasts during homoeostasis and regeneration in the flatworm, Macrostomum lignano. A double immunohistochemical technique was used, performing sequential pulses with the thymidine analogues CldU (chlorodeoxyuridine) and IdU (iododeoxyuridine), separated by variable chase times in the presence of colchicine. Owing to the localized nature of the fluorescent signals (cell nuclei) and variable levels of autofluorescence, standard intensity-based colocalization analyses could not be applied to accurately determine the colocalization. Therefore, an object-based colocalization approach was devised to score the relative number of double-positive cells. Using this approach, T-s (S-phase duration) in the main population of neoblasts was similar to 13 h. During early regeneration, no significant change in T-s was observed.
Keywords
colocalization, cell cycle, neoblast, platyhelminthes, regeneration, S-phase, CYCLE KINETICS, DENTATE GYRUS, REGENERATION, PLANARIAN, DYNAMICS, BROMODEOXYURIDINE, PLATYHELMINTHES, PROLIFERATION, ROOT, IODODEOXYURIDINE

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Citation

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MLA
Verdoodt, Freija et al. “Measurement of S-phase Duration of Adult Stem Cells in the Flatworm Macrostomum Lignano by Double Replication Labelling and Quantitative Colocalization Analysis.” CELL BIOLOGY INTERNATIONAL 36.12 (2012): 1251–1259. Print.
APA
Verdoodt, F., Willems, M., Dhondt, I., Houthoofd, W., Bert, W., & De Vos, W. (2012). Measurement of S-phase duration of adult stem cells in the flatworm Macrostomum lignano by double replication labelling and quantitative colocalization analysis. CELL BIOLOGY INTERNATIONAL, 36(12), 1251–1259.
Chicago author-date
Verdoodt, Freija, Maxime Willems, Ineke Dhondt, Wouter Houthoofd, Wim Bert, and Winnok De Vos. 2012. “Measurement of S-phase Duration of Adult Stem Cells in the Flatworm Macrostomum Lignano by Double Replication Labelling and Quantitative Colocalization Analysis.” Cell Biology International 36 (12): 1251–1259.
Chicago author-date (all authors)
Verdoodt, Freija, Maxime Willems, Ineke Dhondt, Wouter Houthoofd, Wim Bert, and Winnok De Vos. 2012. “Measurement of S-phase Duration of Adult Stem Cells in the Flatworm Macrostomum Lignano by Double Replication Labelling and Quantitative Colocalization Analysis.” Cell Biology International 36 (12): 1251–1259.
Vancouver
1.
Verdoodt F, Willems M, Dhondt I, Houthoofd W, Bert W, De Vos W. Measurement of S-phase duration of adult stem cells in the flatworm Macrostomum lignano by double replication labelling and quantitative colocalization analysis. CELL BIOLOGY INTERNATIONAL. 2012;36(12):1251–9.
IEEE
[1]
F. Verdoodt, M. Willems, I. Dhondt, W. Houthoofd, W. Bert, and W. De Vos, “Measurement of S-phase duration of adult stem cells in the flatworm Macrostomum lignano by double replication labelling and quantitative colocalization analysis,” CELL BIOLOGY INTERNATIONAL, vol. 36, no. 12, pp. 1251–1259, 2012.
@article{3082538,
  abstract     = {Platyhelminthes are highly attractive models for addressing fundamental aspects of stem cell biology in vivo. These organisms possess a unique stem cell system comprised of neoblasts that are the only proliferating cells during adulthood. We have investigated T-s (S-phase duration) of neoblasts during homoeostasis and regeneration in the flatworm, Macrostomum lignano. A double immunohistochemical technique was used, performing sequential pulses with the thymidine analogues CldU (chlorodeoxyuridine) and IdU (iododeoxyuridine), separated by variable chase times in the presence of colchicine. Owing to the localized nature of the fluorescent signals (cell nuclei) and variable levels of autofluorescence, standard intensity-based colocalization analyses could not be applied to accurately determine the colocalization. Therefore, an object-based colocalization approach was devised to score the relative number of double-positive cells. Using this approach, T-s (S-phase duration) in the main population of neoblasts was similar to 13 h. During early regeneration, no significant change in T-s was observed.},
  author       = {Verdoodt, Freija and Willems, Maxime and Dhondt, Ineke and Houthoofd, Wouter and Bert, Wim and De Vos, Winnok},
  issn         = {1065-6995},
  journal      = {CELL BIOLOGY INTERNATIONAL},
  keywords     = {colocalization,cell cycle,neoblast,platyhelminthes,regeneration,S-phase,CYCLE KINETICS,DENTATE GYRUS,REGENERATION,PLANARIAN,DYNAMICS,BROMODEOXYURIDINE,PLATYHELMINTHES,PROLIFERATION,ROOT,IODODEOXYURIDINE},
  language     = {eng},
  number       = {12},
  pages        = {1251--1259},
  title        = {Measurement of S-phase duration of adult stem cells in the flatworm Macrostomum lignano by double replication labelling and quantitative colocalization analysis},
  url          = {http://dx.doi.org/10.1042/CBI20120187},
  volume       = {36},
  year         = {2012},
}

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