Measurement of S-phase duration of adult stem cells in the flatwormMacrostomum lignanoby double replication labelling and quantitative colocalization analysis
| Autor: | Wouter Houthoofd, Maxime Willems, Freija Verdoodt, Winnok H. De Vos, Wim Bert, Ineke Dhondt |
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| Rok vydání: | 2012 |
| Předmět: |
DYNAMICS
S-phase Population PLATYHELMINTHES CYCLE KINETICS PLANARIAN colocalization S Phase chemistry.chemical_compound ROOT REGENERATION Idoxuridine Animals Regeneration education Cell Proliferation neoblast Macrostomum lignano education.field_of_study biology PROLIFERATION Biology and Life Sciences Colocalization Cell Biology General Medicine Cell cycle biology.organism_classification Deoxyuridine Immunohistochemistry BROMODEOXYURIDINE Cell biology Adult Stem Cells DENTATE GYRUS chemistry Platyhelminths Planarian cell cycle Stem cell IODODEOXYURIDINE Bromodeoxyuridine Adult stem cell |
| Zdroj: | CELL BIOLOGY INTERNATIONAL |
| ISSN: | 1095-8355 1065-6995 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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