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BAC transgenic mice to study the expression of P2X2 and P2Y(1) receptors

Extracellular purines are important signaling molecules involved in numerous physiological and pathological processes via the activation of P2 receptors. Information about the spatial and temporal P2 receptor (P2R) expression and its regulation remains crucial for the understanding of the role of P2...

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Autores principales: Grohmann, Marcus, Schumacher, Michaela, Günther, Janka, Singheiser, Stefan M., Nußbaum, Tanja, Wildner, Florian, Gerevich, Zoltan, Jabs, Ronald, Hirnet, Daniela, Lohr, Christian, Illes, Peter, Schmalzing, Günther, Franke, Heike, Hausmann, Ralf
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Netherlands 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8410928/
https://www.ncbi.nlm.nih.gov/pubmed/34050505
http://dx.doi.org/10.1007/s11302-021-09792-9
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author Grohmann, Marcus
Schumacher, Michaela
Günther, Janka
Singheiser, Stefan M.
Nußbaum, Tanja
Wildner, Florian
Gerevich, Zoltan
Jabs, Ronald
Hirnet, Daniela
Lohr, Christian
Illes, Peter
Schmalzing, Günther
Franke, Heike
Hausmann, Ralf
author_facet Grohmann, Marcus
Schumacher, Michaela
Günther, Janka
Singheiser, Stefan M.
Nußbaum, Tanja
Wildner, Florian
Gerevich, Zoltan
Jabs, Ronald
Hirnet, Daniela
Lohr, Christian
Illes, Peter
Schmalzing, Günther
Franke, Heike
Hausmann, Ralf
author_sort Grohmann, Marcus
collection PubMed
description Extracellular purines are important signaling molecules involved in numerous physiological and pathological processes via the activation of P2 receptors. Information about the spatial and temporal P2 receptor (P2R) expression and its regulation remains crucial for the understanding of the role of P2Rs in health and disease. To identify cells carrying P2X2Rs in situ, we have generated BAC transgenic mice that express the P2X2R subunits as fluorescent fusion protein (P2X2-TagRFP). In addition, we generated a BAC P2Y(1)R TagRFP reporter mouse expressing a TagRFP reporter for the P2RY1 gene expression. We demonstrate expression of the P2X2R in a subset of DRG neurons, the brain stem, the hippocampus, as well as on Purkinje neurons of the cerebellum. However, the weak fluorescence intensity in our P2X2R-TagRFP mouse precluded tracking of living cells. Our P2Y(1)R reporter mice confirmed the widespread expression of the P2RY1 gene in the CNS and indicate for the first time P2RY1 gene expression in mouse Purkinje cells, which so far has only been described in rats and humans. Our P2R transgenic models have advanced the understanding of purinergic transmission, but BAC transgenic models appeared not always to be straightforward and permanent reliable. We noticed a loss of fluorescence intensity, which depended on the number of progeny generations. These problems are discussed and may help to provide more successful animal models, even if in future more versatile and adaptable nuclease-mediated genome-editing techniques will be the methods of choice. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s11302-021-09792-9.
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spelling pubmed-84109282021-09-16 BAC transgenic mice to study the expression of P2X2 and P2Y(1) receptors Grohmann, Marcus Schumacher, Michaela Günther, Janka Singheiser, Stefan M. Nußbaum, Tanja Wildner, Florian Gerevich, Zoltan Jabs, Ronald Hirnet, Daniela Lohr, Christian Illes, Peter Schmalzing, Günther Franke, Heike Hausmann, Ralf Purinergic Signal Original Article Extracellular purines are important signaling molecules involved in numerous physiological and pathological processes via the activation of P2 receptors. Information about the spatial and temporal P2 receptor (P2R) expression and its regulation remains crucial for the understanding of the role of P2Rs in health and disease. To identify cells carrying P2X2Rs in situ, we have generated BAC transgenic mice that express the P2X2R subunits as fluorescent fusion protein (P2X2-TagRFP). In addition, we generated a BAC P2Y(1)R TagRFP reporter mouse expressing a TagRFP reporter for the P2RY1 gene expression. We demonstrate expression of the P2X2R in a subset of DRG neurons, the brain stem, the hippocampus, as well as on Purkinje neurons of the cerebellum. However, the weak fluorescence intensity in our P2X2R-TagRFP mouse precluded tracking of living cells. Our P2Y(1)R reporter mice confirmed the widespread expression of the P2RY1 gene in the CNS and indicate for the first time P2RY1 gene expression in mouse Purkinje cells, which so far has only been described in rats and humans. Our P2R transgenic models have advanced the understanding of purinergic transmission, but BAC transgenic models appeared not always to be straightforward and permanent reliable. We noticed a loss of fluorescence intensity, which depended on the number of progeny generations. These problems are discussed and may help to provide more successful animal models, even if in future more versatile and adaptable nuclease-mediated genome-editing techniques will be the methods of choice. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s11302-021-09792-9. Springer Netherlands 2021-05-28 2021-09 /pmc/articles/PMC8410928/ /pubmed/34050505 http://dx.doi.org/10.1007/s11302-021-09792-9 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Original Article
Grohmann, Marcus
Schumacher, Michaela
Günther, Janka
Singheiser, Stefan M.
Nußbaum, Tanja
Wildner, Florian
Gerevich, Zoltan
Jabs, Ronald
Hirnet, Daniela
Lohr, Christian
Illes, Peter
Schmalzing, Günther
Franke, Heike
Hausmann, Ralf
BAC transgenic mice to study the expression of P2X2 and P2Y(1) receptors
title BAC transgenic mice to study the expression of P2X2 and P2Y(1) receptors
title_full BAC transgenic mice to study the expression of P2X2 and P2Y(1) receptors
title_fullStr BAC transgenic mice to study the expression of P2X2 and P2Y(1) receptors
title_full_unstemmed BAC transgenic mice to study the expression of P2X2 and P2Y(1) receptors
title_short BAC transgenic mice to study the expression of P2X2 and P2Y(1) receptors
title_sort bac transgenic mice to study the expression of p2x2 and p2y(1) receptors
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8410928/
https://www.ncbi.nlm.nih.gov/pubmed/34050505
http://dx.doi.org/10.1007/s11302-021-09792-9
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