title: Non-invasive phenotyping and drug testing in single cardiomyocytes or beta-cells by calcium imaging and optogenetics.
abstract: Identification of drug induced electrical instability of the heart curtails development, and introduction, of potentially proarrhythmic drugs. This problem usually requires complimentary contact based approaches such as patch-clamp electrophysiology combined with field stimulation electrodes to observe and control the cell. This produces data with high signal to noise but requires direct physical contact generally preventing high-throughput, or prolonged, phenotyping of single cells or tissues. Combining genetically encoded optogenetic control and spectrally compatible calcium indicator tools into a single adenoviral vector allows the analogous capability for cell control with simultaneous cellular phenotyping without the need for contact. This combination can be applied to single rodent primary adult cardiomyocytes, and human stem cell derived cardiomyocytes, enabling contactless small molecule evaluation for inhibitors of sodium, potassium and calcium channels suggesting it may be useful for early toxicity work. In pancreatic beta-cells it reveals the effects of glucose and the KATP inhibitor gliclazide.
authors.affiliation: Division of Cardiovascular Medicine, Radcliffe Department of Medicine, University of Oxford, Oxford, United Kingdom.
authors.forename: Yu-Fen
authors.initials: YF
authors.lastname: Chang
authors.affiliation: Division of Cardiovascular Medicine, Radcliffe Department of Medicine, University of Oxford, Oxford, United Kingdom.
authors.forename: Connor N
authors.initials: CN
authors.lastname: Broyles
authors.affiliation: Division of Cardiovascular Medicine, Radcliffe Department of Medicine, University of Oxford, Oxford, United Kingdom.
authors.forename: Frances A
authors.initials: FA
authors.lastname: Brook
authors.affiliation: Division of Cardiovascular Medicine, Radcliffe Department of Medicine, University of Oxford, Oxford, United Kingdom.
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Davies
authors.affiliation: Division of Cardiovascular Medicine, Radcliffe Department of Medicine, University of Oxford, Oxford, United Kingdom.
authors.forename: Cameron W
authors.initials: CW
authors.lastname: Turtle
authors.affiliation: Department of Biotechnology, Graduate School of Engineering, Osaka University, Suita, Osaka, Japan.|Department of Biotechnology, School of Engineering, Osaka University, Suita, Osaka, Japan.|Department of Biomolecular Science and Engineering, The Institute of Scientific and Industrial Research, Osaka University, Ibaraki, Osaka, Japan.
authors.forename: Takeharu
authors.initials: T
authors.lastname: Nagai
authors.affiliation: Division of Cardiovascular Medicine, Radcliffe Department of Medicine, University of Oxford, Oxford, United Kingdom.|Department of Cardiology, Oxford University NHS Hospitals Trust, Oxford, United Kingdom.|BHF Centre of Regenerative Medicine, University of Oxford, Oxford, United Kingdom.
authors.forename: Matthew J
authors.identifier: http://orcid.org/0000-0002-9250-6214
authors.initials: MJ
authors.lastname: Daniels
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000378:metabolism; D002478:Cells, Cultured; D005907:Gliclazide / Q000494:pharmacology; D005947:Glucose / Q000378:metabolism; D006168:Guinea Pigs; D006801:Humans; D050417:Insulin-Secreting Cells / Q000187:drug effects* / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000187:drug effects*; D062308:Optogenetics / Q000379:methods; D010641:Phenotype; D011188:Potassium / Q000378:metabolism; D054852:Small Molecule Libraries / Q000494:pharmacology*; D013234:Stem Cells / Q000187:drug effects / Q000378:metabolism
chemical_list: D015220:Calcium Channels; D054852:Small Molecule Libraries; D005907:Gliclazide; D005947:Glucose; D011188:Potassium; D002118:Calcium