Methods for Inferring Cell Cycle Parameters Using Thymidine Analogues

Several procedures have been developed to infer cell-cycle time and the duration of the cycle phases, including tritiated thymidine autoradiography and the immunodetection of thymidine analogues after their incorporation into replicating DNA. These methods have supplied important insights to reveal,...

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Autor: Martí-Clúa, Joaquín|||0000-0002-6774-0092
Tipo de recurso: artículo
Fecha de publicación:2023
País:España
Institución:Universitat Autònoma de Barcelona
Repositorio:Dipòsit Digital de Documents de la UAB
Idioma:inglés
OAI Identifier:oai:ddd.uab.cat:282354
Acceso en línea:https://ddd.uab.cat/record/282354
https://dx.doi.org/urn:doi:10.3390/biology12060885
Access Level:acceso abierto
Palabra clave:Cell cycle parameters
Cell kinetics
S-phase
Thymidine analogues
Toxicity
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spelling Methods for Inferring Cell Cycle Parameters Using Thymidine AnaloguesMartí-Clúa, Joaquín|||0000-0002-6774-0092Cell cycle parametersCell kineticsS-phaseThymidine analoguesToxicitySeveral procedures have been developed to infer cell-cycle time and the duration of the cycle phases, including tritiated thymidine autoradiography and the immunodetection of thymidine analogues after their incorporation into replicating DNA. These methods have supplied important insights to reveal, on fixed tissue sections and using light or electron microscopy, the proliferative behavior of different cell types under different experimental contexts. Thymidine analogous labeling has provided knowledge about cell growth kinetics that would never have been obtained by histological methods alone. Tritiated thymidine autoradiography, 5-bromo-2'-deoxyuridine (BrdU) 5-chloro-2'-deoxyuridine (CldU), 5-iodo-2'-deoxyuridine (IdU), and 5-ethynyl-2'-deoxyiridine (EdU) labeling have been used for identifying the fraction of cells undergoing the S-phase of the cell cycle and to follow the fate of these cells during the embryonic, perinatal, and adult life in several species of vertebrate. In this current review, I will discuss the dosage and times of exposition to the aforementioned thymidine analogues to label most of the cells undergoing the S-phase of the cell cycle. I will also show how to infer, in an asynchronous cell population, the duration of the G, S, and G phases, as well as the growth fraction and the span of the whole cell cycle on the base of some labeling schemes involving a single administration, continuous nucleotide analogue delivery, and double labeling with two thymidine analogues. In this context, the choice of the optimal dose of BrdU, CldU, IdU, and EdU to label S-phase cells is a pivotal aspect to produce neither cytotoxic effects nor alter cell cycle progression. I hope that the information presented in this review can be of use as a reference for researchers involved in the genesis of tissues and organs. 22023-01-0120232023-01-01Articlehttp://purl.org/coar/resource_type/c_6501VoRhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleapplication/pdfhttps://ddd.uab.cat/record/282354https://dx.doi.org/urn:doi:10.3390/biology12060885reponame:Dipòsit Digital de Documents de la UABinstname:Universitat Autònoma de BarcelonaInglésengopen accesshttp://purl.org/coar/access_right/c_abf2Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.https://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:ddd.uab.cat:2823542026-06-06T12:50:31Z
dc.title.none.fl_str_mv Methods for Inferring Cell Cycle Parameters Using Thymidine Analogues
title Methods for Inferring Cell Cycle Parameters Using Thymidine Analogues
spellingShingle Methods for Inferring Cell Cycle Parameters Using Thymidine Analogues
Martí-Clúa, Joaquín|||0000-0002-6774-0092
Cell cycle parameters
Cell kinetics
S-phase
Thymidine analogues
Toxicity
title_short Methods for Inferring Cell Cycle Parameters Using Thymidine Analogues
title_full Methods for Inferring Cell Cycle Parameters Using Thymidine Analogues
title_fullStr Methods for Inferring Cell Cycle Parameters Using Thymidine Analogues
title_full_unstemmed Methods for Inferring Cell Cycle Parameters Using Thymidine Analogues
title_sort Methods for Inferring Cell Cycle Parameters Using Thymidine Analogues
dc.creator.none.fl_str_mv Martí-Clúa, Joaquín|||0000-0002-6774-0092
author Martí-Clúa, Joaquín|||0000-0002-6774-0092
author_facet Martí-Clúa, Joaquín|||0000-0002-6774-0092
author_role author
dc.subject.none.fl_str_mv Cell cycle parameters
Cell kinetics
S-phase
Thymidine analogues
Toxicity
topic Cell cycle parameters
Cell kinetics
S-phase
Thymidine analogues
Toxicity
description Several procedures have been developed to infer cell-cycle time and the duration of the cycle phases, including tritiated thymidine autoradiography and the immunodetection of thymidine analogues after their incorporation into replicating DNA. These methods have supplied important insights to reveal, on fixed tissue sections and using light or electron microscopy, the proliferative behavior of different cell types under different experimental contexts. Thymidine analogous labeling has provided knowledge about cell growth kinetics that would never have been obtained by histological methods alone. Tritiated thymidine autoradiography, 5-bromo-2'-deoxyuridine (BrdU) 5-chloro-2'-deoxyuridine (CldU), 5-iodo-2'-deoxyuridine (IdU), and 5-ethynyl-2'-deoxyiridine (EdU) labeling have been used for identifying the fraction of cells undergoing the S-phase of the cell cycle and to follow the fate of these cells during the embryonic, perinatal, and adult life in several species of vertebrate. In this current review, I will discuss the dosage and times of exposition to the aforementioned thymidine analogues to label most of the cells undergoing the S-phase of the cell cycle. I will also show how to infer, in an asynchronous cell population, the duration of the G, S, and G phases, as well as the growth fraction and the span of the whole cell cycle on the base of some labeling schemes involving a single administration, continuous nucleotide analogue delivery, and double labeling with two thymidine analogues. In this context, the choice of the optimal dose of BrdU, CldU, IdU, and EdU to label S-phase cells is a pivotal aspect to produce neither cytotoxic effects nor alter cell cycle progression. I hope that the information presented in this review can be of use as a reference for researchers involved in the genesis of tissues and organs.
publishDate 2023
dc.date.none.fl_str_mv 2
2023-01-01
2023
2023-01-01
dc.type.none.fl_str_mv Article
http://purl.org/coar/resource_type/c_6501
VoR
http://purl.org/coar/version/c_970fb48d4fbd8a85
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv https://ddd.uab.cat/record/282354
https://dx.doi.org/urn:doi:10.3390/biology12060885
url https://ddd.uab.cat/record/282354
https://dx.doi.org/urn:doi:10.3390/biology12060885
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
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dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
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eu_rights_str_mv openAccess
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dc.source.none.fl_str_mv reponame:Dipòsit Digital de Documents de la UAB
instname:Universitat Autònoma de Barcelona
instname_str Universitat Autònoma de Barcelona
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