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Genetic Labeling of Cells Allows Identification and Tracking of Transgenic Platelets in Mice

Background: The use of knock-out mouse models is crucial to understand platelet activation and aggregation. Methods: Analysis of the global double fluorescent Cre reporter mouse mT/mG that has been crossbred with the megakaryocyte/platelet specific PF4-Cre mouse. Results: Platelets show bright mT (P...

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Autores principales: Krüger, Irena, Reusswig, Friedrich, Krott, Kim Jürgen, Lersch, Celina Fabienne, Spelleken, Martina, Elvers, Margitta
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8037568/
https://www.ncbi.nlm.nih.gov/pubmed/33918229
http://dx.doi.org/10.3390/ijms22073710
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author Krüger, Irena
Reusswig, Friedrich
Krott, Kim Jürgen
Lersch, Celina Fabienne
Spelleken, Martina
Elvers, Margitta
author_facet Krüger, Irena
Reusswig, Friedrich
Krott, Kim Jürgen
Lersch, Celina Fabienne
Spelleken, Martina
Elvers, Margitta
author_sort Krüger, Irena
collection PubMed
description Background: The use of knock-out mouse models is crucial to understand platelet activation and aggregation. Methods: Analysis of the global double fluorescent Cre reporter mouse mT/mG that has been crossbred with the megakaryocyte/platelet specific PF4-Cre mouse. Results: Platelets show bright mT (PF4-Cre negative) and mG (PF4-Cre positive) fluorescence. However, a small proportion of leukocytes was positive for mG fluorescence in PF4-Cre positive mice. In mT/mG;PF4-Cre mice, platelets, and megakaryocytes can be tracked by their specific fluorescence in blood smear, hematopoietic organs and upon thrombus formation. No differences in platelet activation and thrombus formation was observed between mT/mG;PF4-Cre positive and negative mice. Furthermore, hemostasis and in vivo thrombus formation was comparable between genotypes as analyzed by intravital microscopy. Transplantation studies revealed that bone marrow of mT/mG;PF4-Cre mice can be transferred to C57BL/6 mice. Conclusions: The mT/mG Cre reporter mouse is an appropriate model for real-time visualization of platelets, the analysis of cell morphology and the identification of non-recombined platelets. Thus, mT/mG;PF4-Cre mice are important for the analysis of platelet-specific knockout mice. However, a small proportion of leukocytes exhibit mG fluorescence. Therefore, the analysis of platelets beyond hemostasis and thrombosis should be critically evaluated when recombination of immune cells is increased.
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spelling pubmed-80375682021-04-12 Genetic Labeling of Cells Allows Identification and Tracking of Transgenic Platelets in Mice Krüger, Irena Reusswig, Friedrich Krott, Kim Jürgen Lersch, Celina Fabienne Spelleken, Martina Elvers, Margitta Int J Mol Sci Article Background: The use of knock-out mouse models is crucial to understand platelet activation and aggregation. Methods: Analysis of the global double fluorescent Cre reporter mouse mT/mG that has been crossbred with the megakaryocyte/platelet specific PF4-Cre mouse. Results: Platelets show bright mT (PF4-Cre negative) and mG (PF4-Cre positive) fluorescence. However, a small proportion of leukocytes was positive for mG fluorescence in PF4-Cre positive mice. In mT/mG;PF4-Cre mice, platelets, and megakaryocytes can be tracked by their specific fluorescence in blood smear, hematopoietic organs and upon thrombus formation. No differences in platelet activation and thrombus formation was observed between mT/mG;PF4-Cre positive and negative mice. Furthermore, hemostasis and in vivo thrombus formation was comparable between genotypes as analyzed by intravital microscopy. Transplantation studies revealed that bone marrow of mT/mG;PF4-Cre mice can be transferred to C57BL/6 mice. Conclusions: The mT/mG Cre reporter mouse is an appropriate model for real-time visualization of platelets, the analysis of cell morphology and the identification of non-recombined platelets. Thus, mT/mG;PF4-Cre mice are important for the analysis of platelet-specific knockout mice. However, a small proportion of leukocytes exhibit mG fluorescence. Therefore, the analysis of platelets beyond hemostasis and thrombosis should be critically evaluated when recombination of immune cells is increased. MDPI 2021-04-02 /pmc/articles/PMC8037568/ /pubmed/33918229 http://dx.doi.org/10.3390/ijms22073710 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Krüger, Irena
Reusswig, Friedrich
Krott, Kim Jürgen
Lersch, Celina Fabienne
Spelleken, Martina
Elvers, Margitta
Genetic Labeling of Cells Allows Identification and Tracking of Transgenic Platelets in Mice
title Genetic Labeling of Cells Allows Identification and Tracking of Transgenic Platelets in Mice
title_full Genetic Labeling of Cells Allows Identification and Tracking of Transgenic Platelets in Mice
title_fullStr Genetic Labeling of Cells Allows Identification and Tracking of Transgenic Platelets in Mice
title_full_unstemmed Genetic Labeling of Cells Allows Identification and Tracking of Transgenic Platelets in Mice
title_short Genetic Labeling of Cells Allows Identification and Tracking of Transgenic Platelets in Mice
title_sort genetic labeling of cells allows identification and tracking of transgenic platelets in mice
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8037568/
https://www.ncbi.nlm.nih.gov/pubmed/33918229
http://dx.doi.org/10.3390/ijms22073710
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