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GNrep mouse: A reporter mouse for front–rear cell polarity

Cell migration is essential during development, regeneration, homeostasis, and disease. Depending on the microenvironment, cells use different mechanisms to migrate. Yet, all modes of migration require the establishment of an intracellular front–rear polarity axis for directional movement. Although...

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Detalles Bibliográficos
Autores principales: Barbacena, Pedro, Ouarné, Marie, Haigh, Jody J., Vasconcelos, Francisca F., Pezzarossa, Anna, Franco, Claudio A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley & Sons, Inc. 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6618267/
https://www.ncbi.nlm.nih.gov/pubmed/30990965
http://dx.doi.org/10.1002/dvg.23299
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author Barbacena, Pedro
Ouarné, Marie
Haigh, Jody J.
Vasconcelos, Francisca F.
Pezzarossa, Anna
Franco, Claudio A.
author_facet Barbacena, Pedro
Ouarné, Marie
Haigh, Jody J.
Vasconcelos, Francisca F.
Pezzarossa, Anna
Franco, Claudio A.
author_sort Barbacena, Pedro
collection PubMed
description Cell migration is essential during development, regeneration, homeostasis, and disease. Depending on the microenvironment, cells use different mechanisms to migrate. Yet, all modes of migration require the establishment of an intracellular front–rear polarity axis for directional movement. Although front–rear polarity can be easily identified in in vitro conditions, its assessment in vivo by live‐imaging is challenging due to tissue complexity and lack of reliable markers. Here, we describe a novel and unique double fluorescent reporter mouse line to study front–rear cell polarity in living tissues, called GNrep. This mouse line simultaneously labels Golgi complexes and nuclei allowing the assignment of a nucleus‐to‐Golgi axis to each cell, which functions as a readout for cell front–rear polarity. As a proof‐of‐principle, we validated the efficiency of the GNrep line using an endothelial‐specific Cre mouse line. We show that the GNrep labels the nucleus and the Golgi apparatus of endothelial cells with very high efficiency and high specificity. Importantly, the features of fluorescent intensity and localization for both mCherry and eGFP fluorescent intensity and localization allow automated segmentation and assignment of polarity vectors in complex tissues, making GNrep a great tool to study cell behavior in large‐scale automated analyses. Altogether, the GNrep mouse line, in combination with different Cre recombinase lines, is a novel and unique tool to study of front–rear polarity in mice, both in fixed tissues or in intravital live imaging. This new line will be instrumental to understand cell migration and polarity in development, homeostasis, and disease.
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spelling pubmed-66182672019-07-22 GNrep mouse: A reporter mouse for front–rear cell polarity Barbacena, Pedro Ouarné, Marie Haigh, Jody J. Vasconcelos, Francisca F. Pezzarossa, Anna Franco, Claudio A. Genesis Technology Reports Cell migration is essential during development, regeneration, homeostasis, and disease. Depending on the microenvironment, cells use different mechanisms to migrate. Yet, all modes of migration require the establishment of an intracellular front–rear polarity axis for directional movement. Although front–rear polarity can be easily identified in in vitro conditions, its assessment in vivo by live‐imaging is challenging due to tissue complexity and lack of reliable markers. Here, we describe a novel and unique double fluorescent reporter mouse line to study front–rear cell polarity in living tissues, called GNrep. This mouse line simultaneously labels Golgi complexes and nuclei allowing the assignment of a nucleus‐to‐Golgi axis to each cell, which functions as a readout for cell front–rear polarity. As a proof‐of‐principle, we validated the efficiency of the GNrep line using an endothelial‐specific Cre mouse line. We show that the GNrep labels the nucleus and the Golgi apparatus of endothelial cells with very high efficiency and high specificity. Importantly, the features of fluorescent intensity and localization for both mCherry and eGFP fluorescent intensity and localization allow automated segmentation and assignment of polarity vectors in complex tissues, making GNrep a great tool to study cell behavior in large‐scale automated analyses. Altogether, the GNrep mouse line, in combination with different Cre recombinase lines, is a novel and unique tool to study of front–rear polarity in mice, both in fixed tissues or in intravital live imaging. This new line will be instrumental to understand cell migration and polarity in development, homeostasis, and disease. John Wiley & Sons, Inc. 2019-04-16 2019-06 /pmc/articles/PMC6618267/ /pubmed/30990965 http://dx.doi.org/10.1002/dvg.23299 Text en © 2019 The Authors. Genesis published by Wiley Periodicals, Inc. This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.
spellingShingle Technology Reports
Barbacena, Pedro
Ouarné, Marie
Haigh, Jody J.
Vasconcelos, Francisca F.
Pezzarossa, Anna
Franco, Claudio A.
GNrep mouse: A reporter mouse for front–rear cell polarity
title GNrep mouse: A reporter mouse for front–rear cell polarity
title_full GNrep mouse: A reporter mouse for front–rear cell polarity
title_fullStr GNrep mouse: A reporter mouse for front–rear cell polarity
title_full_unstemmed GNrep mouse: A reporter mouse for front–rear cell polarity
title_short GNrep mouse: A reporter mouse for front–rear cell polarity
title_sort gnrep mouse: a reporter mouse for front–rear cell polarity
topic Technology Reports
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6618267/
https://www.ncbi.nlm.nih.gov/pubmed/30990965
http://dx.doi.org/10.1002/dvg.23299
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