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Generation of a Homozygous Transgenic Rat Strain Stably Expressing a Calcium Sensor Protein for Direct Examination of Calcium Signaling
In drug discovery, prediction of selectivity and toxicity require the evaluation of cellular calcium homeostasis. The rat is a preferred laboratory animal for pharmacology and toxicology studies, while currently no calcium indicator protein expressing rat model is available. We established a transge...
Autores principales: | , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4522653/ https://www.ncbi.nlm.nih.gov/pubmed/26234466 http://dx.doi.org/10.1038/srep12645 |
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author | Szebényi, Kornélia Füredi, András Kolacsek, Orsolya Pergel, Enikő Bősze, Zsuzsanna Bender, Balázs Vajdovich, Péter Tóvári, József Homolya, László Szakács, Gergely Héja, László Enyedi, Ágnes Sarkadi, Balázs Apáti, Ágota Orbán, Tamás I. |
author_facet | Szebényi, Kornélia Füredi, András Kolacsek, Orsolya Pergel, Enikő Bősze, Zsuzsanna Bender, Balázs Vajdovich, Péter Tóvári, József Homolya, László Szakács, Gergely Héja, László Enyedi, Ágnes Sarkadi, Balázs Apáti, Ágota Orbán, Tamás I. |
author_sort | Szebényi, Kornélia |
collection | PubMed |
description | In drug discovery, prediction of selectivity and toxicity require the evaluation of cellular calcium homeostasis. The rat is a preferred laboratory animal for pharmacology and toxicology studies, while currently no calcium indicator protein expressing rat model is available. We established a transgenic rat strain stably expressing the GCaMP2 fluorescent calcium sensor by a transposon-based methodology. Zygotes were co-injected with mRNA of transposase and a CAG-GCaMP2 expressing construct, and animals with one transgene copy were pre-selected by measuring fluorescence in blood cells. A homozygous rat strain was generated with high sensor protein expression in the heart, kidney, liver, and blood cells. No pathological alterations were found in these animals, and fluorescence measurements in cardiac tissue slices and primary cultures demonstrated the applicability of this system for studying calcium signaling. We show here that the GCaMP2 expressing rat cardiomyocytes allow the prediction of cardiotoxic drug side-effects, and provide evidence for the role of Na(+)/Ca(2+) exchanger and its beneficial pharmacological modulation in cardiac reperfusion. Our data indicate that drug-induced alterations and pathological processes can be followed by using this rat model, suggesting that transgenic rats expressing a calcium-sensitive protein provide a valuable system for pharmacological and toxicological studies. |
format | Online Article Text |
id | pubmed-4522653 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-45226532015-08-06 Generation of a Homozygous Transgenic Rat Strain Stably Expressing a Calcium Sensor Protein for Direct Examination of Calcium Signaling Szebényi, Kornélia Füredi, András Kolacsek, Orsolya Pergel, Enikő Bősze, Zsuzsanna Bender, Balázs Vajdovich, Péter Tóvári, József Homolya, László Szakács, Gergely Héja, László Enyedi, Ágnes Sarkadi, Balázs Apáti, Ágota Orbán, Tamás I. Sci Rep Article In drug discovery, prediction of selectivity and toxicity require the evaluation of cellular calcium homeostasis. The rat is a preferred laboratory animal for pharmacology and toxicology studies, while currently no calcium indicator protein expressing rat model is available. We established a transgenic rat strain stably expressing the GCaMP2 fluorescent calcium sensor by a transposon-based methodology. Zygotes were co-injected with mRNA of transposase and a CAG-GCaMP2 expressing construct, and animals with one transgene copy were pre-selected by measuring fluorescence in blood cells. A homozygous rat strain was generated with high sensor protein expression in the heart, kidney, liver, and blood cells. No pathological alterations were found in these animals, and fluorescence measurements in cardiac tissue slices and primary cultures demonstrated the applicability of this system for studying calcium signaling. We show here that the GCaMP2 expressing rat cardiomyocytes allow the prediction of cardiotoxic drug side-effects, and provide evidence for the role of Na(+)/Ca(2+) exchanger and its beneficial pharmacological modulation in cardiac reperfusion. Our data indicate that drug-induced alterations and pathological processes can be followed by using this rat model, suggesting that transgenic rats expressing a calcium-sensitive protein provide a valuable system for pharmacological and toxicological studies. Nature Publishing Group 2015-08-03 /pmc/articles/PMC4522653/ /pubmed/26234466 http://dx.doi.org/10.1038/srep12645 Text en Copyright © 2015, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Article Szebényi, Kornélia Füredi, András Kolacsek, Orsolya Pergel, Enikő Bősze, Zsuzsanna Bender, Balázs Vajdovich, Péter Tóvári, József Homolya, László Szakács, Gergely Héja, László Enyedi, Ágnes Sarkadi, Balázs Apáti, Ágota Orbán, Tamás I. Generation of a Homozygous Transgenic Rat Strain Stably Expressing a Calcium Sensor Protein for Direct Examination of Calcium Signaling |
title | Generation of a Homozygous Transgenic Rat Strain Stably Expressing a Calcium Sensor Protein for Direct Examination of Calcium Signaling |
title_full | Generation of a Homozygous Transgenic Rat Strain Stably Expressing a Calcium Sensor Protein for Direct Examination of Calcium Signaling |
title_fullStr | Generation of a Homozygous Transgenic Rat Strain Stably Expressing a Calcium Sensor Protein for Direct Examination of Calcium Signaling |
title_full_unstemmed | Generation of a Homozygous Transgenic Rat Strain Stably Expressing a Calcium Sensor Protein for Direct Examination of Calcium Signaling |
title_short | Generation of a Homozygous Transgenic Rat Strain Stably Expressing a Calcium Sensor Protein for Direct Examination of Calcium Signaling |
title_sort | generation of a homozygous transgenic rat strain stably expressing a calcium sensor protein for direct examination of calcium signaling |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4522653/ https://www.ncbi.nlm.nih.gov/pubmed/26234466 http://dx.doi.org/10.1038/srep12645 |
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