{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T11:22:27Z","timestamp":1792668147717,"version":"4.5.0"},"reference-count":53,"publisher":"Ovid Technologies (Wolters Kluwer Health)","issue":"8","content-domain":{"domain":["www.ahajournals.org"],"crossmark-restriction":true},"short-container-title":["Circulation Research"],"published-print":{"date-parts":[[2019,4,12]]},"abstract":"<jats:sec>\n                    <jats:title>\n                      <jats:underline>Rationale:<\/jats:underline>\n                    <\/jats:title>\n                    <jats:p>\n                      Subcellular Ca\n                      <jats:sup>2+<\/jats:sup>\n                      indicators have yet to be developed for the myofilament where disease mutation or small molecules may alter contractility through myofilament Ca\n                      <jats:sup>2+<\/jats:sup>\n                      sensitivity. Here, we develop and characterize genetically encoded Ca\n                      <jats:sup>2+<\/jats:sup>\n                      indicators restricted to the myofilament to directly visualize Ca\n                      <jats:sup>2+<\/jats:sup>\n                      changes in the sarcomere.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>\n                      <jats:underline>Objective:<\/jats:underline>\n                    <\/jats:title>\n                    <jats:p>\n                      To produce and validate myofilament-restricted Ca\n                      <jats:sup>2+<\/jats:sup>\n                      imaging probes in an adenoviral transduction adult cardiomyocyte model using drugs that alter myofilament function (MYK-461, omecamtiv mecarbil, and levosimendan) or following cotransduction of 2 established hypertrophic cardiomyopathy disease-causing mutants (cTnT [Troponin T] R92Q and cTnI [Troponin I] R145G) that alter myofilament Ca\n                      <jats:sup>2+<\/jats:sup>\n                      handling.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>\n                      <jats:underline>Methods and Results:<\/jats:underline>\n                    <\/jats:title>\n                    <jats:p>\n                      When expressed in adult ventricular cardiomyocytes RGECO-TnT (Troponin T)\/TnI (Troponin I) sensors localize correctly to the sarcomere without contractile impairment. Both sensors report cyclical changes in fluorescence in paced cardiomyocytes with reduced Ca\n                      <jats:sup>2+<\/jats:sup>\n                      on and increased Ca\n                      <jats:sup>2+<\/jats:sup>\n                      off rates compared with unconjugated RGECO. RGECO-TnT\/TnI revealed changes to localized Ca\n                      <jats:sup>2+<\/jats:sup>\n                      handling conferred by MYK-461 and levosimendan, including an increase in Ca\n                      <jats:sup>2+<\/jats:sup>\n                      binding rates with both levosimendan and MYK-461 not detected by an unrestricted protein sensor. Coadenoviral transduction of RGECO-TnT\/TnI with hypertrophic cardiomyopathy causing thin filament mutants showed that the mutations increase myofilament [Ca\n                      <jats:sup>2+<\/jats:sup>\n                      ] in systole, lengthen time to peak systolic [Ca\n                      <jats:sup>2+<\/jats:sup>\n                      ], and delay [Ca\n                      <jats:sup>2+<\/jats:sup>\n                      ] release. This contrasts with the effect of the same mutations on cytoplasmic Ca\n                      <jats:sup>2+<\/jats:sup>\n                      , when measured using unrestricted RGECO where changes to peak systolic Ca\n                      <jats:sup>2+<\/jats:sup>\n                      are inconsistent between the 2 mutations. These data contrast with previous findings using chemical dyes that show no alteration of [Ca\n                      <jats:sup>2+<\/jats:sup>\n                      ] transient amplitude or time to peak Ca\n                      <jats:sup>2+<\/jats:sup>\n                      .\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>\n                      <jats:underline>Conclusions:<\/jats:underline>\n                    <\/jats:title>\n                    <jats:p>\n                      RGECO-TnT\/TnI are functionally equivalent. They visualize Ca\n                      <jats:sup>2+<\/jats:sup>\n                      within the myofilament and reveal unrecognized aspects of small molecule and disease-associated mutations in living cells.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1161\/circresaha.118.314600","type":"journal-article","created":{"date-parts":[[2019,2,8]],"date-time":"2019-02-08T08:53:25Z","timestamp":1549616005000},"page":"1228-1239","update-policy":"https:\/\/doi.org\/10.1161\/crossmarkpolicy","source":"Crossref","is-referenced-by-count":54,"title":["Measurement of Myofilament-Localized Calcium Dynamics in Adult Cardiomyocytes and the Effect of Hypertrophic Cardiomyopathy Mutations"],"prefix":"10.1161","volume":"124","author":[{"given":"Alexander J.","family":"Sparrow","sequence":"first","affiliation":[{"name":"From the Division of Cardiovascular Medicine, Radcliffe Department of Medicine (A.J.S., K.S., S.P., Y.-F.C., C.N.B., F.A.B., H.W., C.S.R., P.R., M.J.D.), University of Oxford, United Kingdom"},{"name":"BHF Centre of Research Excellence (A.J.S., S.P., Y.-F.C., C.N.B., F.A.B., H.W., C.S.R., P.R., M.J.D.), University of Oxford, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kolja","family":"Sievert","sequence":"additional","affiliation":[{"name":"From the Division of Cardiovascular Medicine, Radcliffe Department of Medicine (A.J.S., K.S., S.P., Y.-F.C., C.N.B., F.A.B., H.W., C.S.R., P.R., M.J.D.), University of Oxford, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Suketu","family":"Patel","sequence":"additional","affiliation":[{"name":"From the Division of Cardiovascular Medicine, Radcliffe Department of Medicine (A.J.S., K.S., S.P., Y.-F.C., C.N.B., F.A.B., H.W., C.S.R., P.R., M.J.D.), University of Oxford, United Kingdom"},{"name":"BHF Centre of Research Excellence (A.J.S., S.P., Y.-F.C., C.N.B., F.A.B., H.W., C.S.R., P.R., M.J.D.), University of Oxford, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu-Fen","family":"Chang","sequence":"additional","affiliation":[{"name":"From the Division of Cardiovascular Medicine, Radcliffe Department of Medicine (A.J.S., K.S., S.P., Y.-F.C., C.N.B., F.A.B., H.W., C.S.R., P.R., M.J.D.), University of Oxford, United Kingdom"},{"name":"BHF Centre of Research Excellence (A.J.S., S.P., Y.-F.C., C.N.B., F.A.B., H.W., C.S.R., P.R., M.J.D.), University of Oxford, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Connor N.","family":"Broyles","sequence":"additional","affiliation":[{"name":"From the Division of Cardiovascular Medicine, Radcliffe Department of Medicine (A.J.S., K.S., S.P., Y.-F.C., C.N.B., F.A.B., H.W., C.S.R., P.R., M.J.D.), University of Oxford, United Kingdom"},{"name":"BHF Centre of Research Excellence (A.J.S., S.P., Y.-F.C., C.N.B., F.A.B., H.W., C.S.R., P.R., M.J.D.), University of Oxford, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Frances A.","family":"Brook","sequence":"additional","affiliation":[{"name":"From the Division of Cardiovascular Medicine, Radcliffe Department of Medicine (A.J.S., K.S., S.P., Y.-F.C., C.N.B., F.A.B., H.W., C.S.R., P.R., M.J.D.), University of Oxford, United Kingdom"},{"name":"BHF Centre of Research Excellence (A.J.S., S.P., Y.-F.C., C.N.B., F.A.B., H.W., C.S.R., P.R., M.J.D.), University of Oxford, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hugh","family":"Watkins","sequence":"additional","affiliation":[{"name":"From the Division of Cardiovascular Medicine, Radcliffe Department of Medicine (A.J.S., K.S., S.P., Y.-F.C., C.N.B., F.A.B., H.W., C.S.R., P.R., M.J.D.), University of Oxford, United Kingdom"},{"name":"BHF Centre of Research Excellence (A.J.S., S.P., Y.-F.C., C.N.B., F.A.B., H.W., C.S.R., P.R., M.J.D.), University of Oxford, United Kingdom"},{"name":"Department of Cardiology, Oxford University NHS Hospitals Trust, United Kingdom (H.W., M.J.D.)"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael A.","family":"Geeves","sequence":"additional","affiliation":[{"name":"Department of Biosciences, University of Kent, Canterbury, United Kingdom (M.A.G.)"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Charles S.","family":"Redwood","sequence":"additional","affiliation":[{"name":"From the Division of Cardiovascular Medicine, Radcliffe Department of Medicine (A.J.S., K.S., S.P., Y.-F.C., C.N.B., F.A.B., H.W., C.S.R., P.R., M.J.D.), University of Oxford, United Kingdom"},{"name":"BHF Centre of Research Excellence (A.J.S., S.P., Y.-F.C., C.N.B., F.A.B., H.W., C.S.R., P.R., M.J.D.), University of Oxford, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paul","family":"Robinson","sequence":"additional","affiliation":[{"name":"From the Division of Cardiovascular Medicine, Radcliffe Department of Medicine (A.J.S., K.S., S.P., Y.-F.C., C.N.B., F.A.B., H.W., C.S.R., P.R., M.J.D.), University of Oxford, United Kingdom"},{"name":"BHF Centre of Research Excellence (A.J.S., S.P., Y.-F.C., C.N.B., F.A.B., H.W., C.S.R., P.R., M.J.D.), University of Oxford, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matthew J.","family":"Daniels","sequence":"additional","affiliation":[{"name":"From the Division of Cardiovascular Medicine, Radcliffe Department of Medicine (A.J.S., K.S., S.P., Y.-F.C., C.N.B., F.A.B., H.W., C.S.R., P.R., M.J.D.), University of Oxford, United Kingdom"},{"name":"BHF Centre of Research Excellence (A.J.S., S.P., Y.-F.C., C.N.B., F.A.B., H.W., C.S.R., P.R., M.J.D.), University of Oxford, United Kingdom"},{"name":"BHF Centre of Regenerative Medicine (M.J.D.), University of Oxford, United Kingdom"},{"name":"Department of Cardiology, Oxford University NHS Hospitals Trust, United Kingdom (H.W., M.J.D.)"},{"name":"Department of Biotechnology, Graduate School of Engineering, Osaka University, Suita, Japan 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