title: Generation of a Homozygous Transgenic Rat Strain Stably Expressing a Calcium Sensor Protein for Direct Examination of Calcium Signaling.
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(+)/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.
authors.affiliation: Institute of Enzymology, Research Centre for Natural Sciences, Hungarian Academy of Sciences, Budapest, Hungary.
authors.forename: Kornélia
authors.initials: K
authors.lastname: Szebényi
authors.affiliation: Institute of Enzymology, Research Centre for Natural Sciences, Hungarian Academy of Sciences, Budapest, Hungary.
authors.forename: András
authors.initials: A
authors.lastname: Füredi
authors.affiliation: Institute of Enzymology, Research Centre for Natural Sciences, Hungarian Academy of Sciences, Budapest, Hungary.
authors.forename: Orsolya
authors.initials: O
authors.lastname: Kolacsek
authors.affiliation: Institute of Enzymology, Research Centre for Natural Sciences, Hungarian Academy of Sciences, Budapest, Hungary.
authors.forename: Enikő
authors.initials: E
authors.lastname: Pergel
authors.affiliation: NARIC-ABC, Gödöllő, Hungary.
authors.forename: Zsuzsanna
authors.initials: Z
authors.lastname: Bősze
authors.affiliation: ImmunoGenes Ltd., Budakeszi, Hungary.
authors.forename: Balázs
authors.initials: B
authors.lastname: Bender
authors.affiliation: Department of Clinical Pathology and Oncology, Faculty of Veterinary Science, Szent István University, Budapest, Hungary.
authors.forename: Péter
authors.initials: P
authors.lastname: Vajdovich
authors.affiliation: Department of Experimental Pharmacology, National Institute of Oncology, Budapest, Hungary.
authors.forename: József
authors.initials: J
authors.lastname: Tóvári
authors.affiliation: Institute of Enzymology, Research Centre for Natural Sciences, Hungarian Academy of Sciences, Budapest, Hungary.
authors.forename: László
authors.initials: L
authors.lastname: Homolya
authors.affiliation: Institute of Enzymology, Research Centre for Natural Sciences, Hungarian Academy of Sciences, Budapest, Hungary.
authors.forename: Gergely
authors.initials: G
authors.lastname: Szakács
authors.affiliation: Institute of Organic Chemistry, Research Centre for Natural Sciences, Hungarian Academy of Sciences, Budapest, Hungary.
authors.forename: László
authors.initials: L
authors.lastname: Héja
authors.affiliation: 2nd Institute of Pathology, Semmelweis University, Budapest, Hungary.
authors.forename: Ágnes
authors.initials: Á
authors.lastname: Enyedi
authors.affiliation: 1] Institute of Enzymology, Research Centre for Natural Sciences, Hungarian Academy of Sciences, Budapest, Hungary [2] Department of Biophysics and Radiation Biology, Semmelweis University, Budapest, Hungary and MTA-SE Molecular Biophysics Research Group, Budapest, Hungary.
authors.forename: Balázs
authors.initials: B
authors.lastname: Sarkadi
authors.affiliation: Institute of Enzymology, Research Centre for Natural Sciences, Hungarian Academy of Sciences, Budapest, Hungary.
authors.forename: Ágota
authors.initials: Á
authors.lastname: Apáti
authors.affiliation: Institute of Enzymology, Research Centre for Natural Sciences, Hungarian Academy of Sciences, Budapest, Hungary.
authors.forename: Tamás I
authors.initials: TI
authors.lastname: Orbán
mesh_terms: D000818:Animals; D020013:Calcium Signaling / Q000187:drug effects / Q000235:genetics; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2 / Q000235:genetics* / Q000378:metabolism; D015687:Cell Hypoxia / Q000187:drug effects; D002478:Cells, Cultured; D005260:Female; D005818:Genetic Engineering / Q000379:methods; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D006720:Homozygote; D008297:Male; D015767:Mefloquine / Q000494:pharmacology; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000378:metabolism; D011401:Promoter Regions, Genetic; D017207:Rats, Sprague-Dawley; D055647:Rats, Transgenic / Q000235:genetics*; D017208:Rats, Wistar; D013890:Thiourea / Q000031:analogs & derivatives / Q000494:pharmacology
chemical_list: C101670:2-(2-(4-(4-nitrobenzyloxy)phenyl)ethyl)isothiourea methanesulfonate; D049452:Green Fluorescent Proteins; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D013890:Thiourea; D015767:Mefloquine