Generation of a Homozygous Transgenic Rat Strain Stably Expressing a Calcium Sensor Protein for Direct Examination of Calcium Signaling

Kornelia Szebenyi, Andras Füredi, Orsolya Kolacsek, Enik Pergel, Z. Bösze, Balazs Bender, Peter Vajdovich, J. Tóvári, L. Homolya, G. Szakács, Laszlo Heja, A. Enyedi, B. Sarkadi, A. Apáti, Tamas I. Orban

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Abstract

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+/Ca2+ 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.

Original languageEnglish
Article number12645
JournalScientific Reports
Volume5
DOIs
Publication statusPublished - Aug 3 2015

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Transgenic Rats
Calcium Signaling
Calcium
Pharmacology
Proteins
Toxicology
Blood Cells
Fluorescence
Transposases
Zygote
Laboratory Animals
Pathologic Processes
Drug Discovery
Drug-Related Side Effects and Adverse Reactions
Transgenes
Cardiac Myocytes
Reperfusion
Homeostasis
Kidney
Messenger RNA

ASJC Scopus subject areas

  • General

Cite this

Generation of a Homozygous Transgenic Rat Strain Stably Expressing a Calcium Sensor Protein for Direct Examination of Calcium Signaling. / Szebenyi, Kornelia; Füredi, Andras; Kolacsek, Orsolya; Pergel, Enik; Bösze, Z.; Bender, Balazs; Vajdovich, Peter; Tóvári, J.; Homolya, L.; Szakács, G.; Heja, Laszlo; Enyedi, A.; Sarkadi, B.; Apáti, A.; Orban, Tamas I.

In: Scientific Reports, Vol. 5, 12645, 03.08.2015.

Research output: Contribution to journalArticle

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AU - Pergel, Enik

AU - Bösze, Z.

AU - Bender, Balazs

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AU - Tóvári, J.

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AU - Szakács, G.

AU - Heja, Laszlo

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