{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,26]],"date-time":"2026-06-26T11:47:00Z","timestamp":1782474420746,"version":"3.54.5"},"reference-count":56,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2026,5,30]],"date-time":"2026-05-30T00:00:00Z","timestamp":1780099200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"},{"start":{"date-parts":[[2026,5,30]],"date-time":"2026-05-30T00:00:00Z","timestamp":1780099200000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Current Protocols"],"published-print":{"date-parts":[[2026,6]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Mitochondrial calcium (mCa\n                    <jats:sup>2+<\/jats:sup>\n                    ) homeostasis promotes oxidative metabolism within the physiological range; however, dysregulation can trigger necrotic cell death in diseases such as cardiac ischemia\u2010reperfusion injury, muscular dystrophy, and neurodegenerative disorders, including Alzheimer's disease. It is widely understood that mitochondria exhibit rapid Ca\n                    <jats:sup>2+<\/jats:sup>\n                    uptake primarily mediated by the mitochondrial calcium uniporter (MCU) complex, and that Ca\n                    <jats:sup>2+<\/jats:sup>\n                    is exported via a combination of Na\n                    <jats:sup>+<\/jats:sup>\n                    \/Ca\n                    <jats:sup>2+<\/jats:sup>\n                    and H\n                    <jats:sup>+<\/jats:sup>\n                    \/Ca\n                    <jats:sup>2+<\/jats:sup>\n                    exchange processes. However, the proteins which mediate mCa\n                    <jats:sup>2+<\/jats:sup>\n                    transport have only been partially identified. A particular challenge in determining which proteins mediate mCa\n                    <jats:sup>2+<\/jats:sup>\n                    efflux and their relative contributions to mCa\n                    <jats:sup>2+<\/jats:sup>\n                    homeostasis is the lack of a clear, reproducible assay for mCa\n                    <jats:sup>2+<\/jats:sup>\n                    efflux applicable across genotypes. Here, we provide instructions for an optimized fluorometric method to measure mCa\n                    <jats:sup>2+<\/jats:sup>\n                    efflux in isolated mitochondria that establishes robust Ca\n                    <jats:sup>2+<\/jats:sup>\n                    efflux signals, differentiates between total and Na\n                    <jats:sup>+<\/jats:sup>\n                    \u2010independent Ca\n                    <jats:sup>2+<\/jats:sup>\n                    efflux modalities, and generates highly reproducible data, allowing comparisons across tissues and genotypes. \u00a9 2026 The Author(s). Current Protocols published by Wiley Periodicals LLC.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 1<\/jats:bold>\n                    : Total Calcium Efflux Assay in Isolated Mitochondria\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Alternate Protocol 1<\/jats:bold>\n                    : Na\n                    <jats:sup>+<\/jats:sup>\n                    \u2010Independent Calcium Efflux Assay in Isolated Mitochondria\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Support Protocol 1<\/jats:bold>\n                    : Isolation of Cardiac Mitochondria\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Support Protocol 2<\/jats:bold>\n                    : Isolation of Skeletal Muscle Mitochondria\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Support Protocol 3<\/jats:bold>\n                    : Isolation of Liver Mitochondria\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Support Protocol 4<\/jats:bold>\n                    : Isolation of Cell Line Mitochondria\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 2<\/jats:bold>\n                    : Analysis of Calcium Efflux\n                  <\/jats:p>","DOI":"10.1002\/cpz1.70393","type":"journal-article","created":{"date-parts":[[2026,5,30]],"date-time":"2026-05-30T14:35:41Z","timestamp":1780151741000},"source":"Crossref","is-referenced-by-count":0,"title":["Measuring Calcium Efflux in Isolated Primary Mitochondria"],"prefix":"10.1002","volume":"6","author":[{"given":"Ta'Aliyah M.","family":"Jones","sequence":"first","affiliation":[{"name":"Department of Integrative Physiology Baylor College of Medicine  Houston TX USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stephen","family":"Hurst","sequence":"additional","affiliation":[{"name":"Department of Integrative Physiology Baylor College of Medicine  Houston TX USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael J.","family":"Bround","sequence":"additional","affiliation":[{"name":"Department of Integrative Physiology Baylor College of Medicine  Houston TX USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,5,30]]},"reference":[{"key":"e_1_2_12_2_1_1","doi-asserted-by":"publisher","DOI":"10.1152\/ajpcell.00502.2020"},{"key":"e_1_2_12_2_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.yjmcc.2018.12.019"},{"key":"e_1_2_12_2_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbc.2023.104708"},{"key":"e_1_2_12_2_4_1","doi-asserted-by":"publisher","DOI":"10.15252\/embr.202254978"},{"key":"e_1_2_12_2_5_1","doi-asserted-by":"publisher","DOI":"10.3389\/fphys.2017.00839"},{"key":"e_1_2_12_2_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/0014\u20105793(82)80476\u20106"},{"key":"e_1_2_12_2_7_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1432\u20101033.1983.tb07578.x"},{"key":"e_1_2_12_2_8_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598\u2010024\u201057340\u20103"},{"key":"e_1_2_12_2_9_1","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.adi2767"},{"key":"e_1_2_12_2_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/0022\u20102828(74)90077\u20107"},{"key":"e_1_2_12_2_11_1","doi-asserted-by":"publisher","DOI":"10.2337\/db20\u20100779"},{"key":"e_1_2_12_2_12_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1432\u20101033.1977.tb11839.x"},{"key":"e_1_2_12_2_13_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M113.540898"},{"key":"e_1_2_12_2_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbabio.2009.01.005"},{"key":"e_1_2_12_2_15_1","doi-asserted-by":"publisher","DOI":"10.1096\/fj.14\u2010256453"},{"key":"e_1_2_12_2_16_1","doi-asserted-by":"publisher","DOI":"10.1038\/nm.1868"},{"key":"e_1_2_12_2_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.neurobiolaging.2009.03.003"},{"key":"e_1_2_12_2_18_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0702228104"},{"key":"e_1_2_12_2_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbamem.2005.02.018"},{"key":"e_1_2_12_2_20_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.yjmcc.2022.03.001"},{"key":"e_1_2_12_2_21_1","doi-asserted-by":"publisher","DOI":"10.1038\/s42255\u2010025\u201001250\u20109"},{"key":"e_1_2_12_2_22_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41418\u2010018\u20100191\u20107"},{"key":"e_1_2_12_2_23_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi2018909"},{"key":"e_1_2_12_2_24_1","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCRESAHA.119.316369"},{"key":"e_1_2_12_2_25_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tem.2024.04.005"},{"key":"e_1_2_12_2_26_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2023.113465"},{"key":"e_1_2_12_2_27_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1175145"},{"key":"e_1_2_12_2_28_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1308558110"},{"key":"e_1_2_12_2_29_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ajhg.2022.07.007"},{"key":"e_1_2_12_2_30_1","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.aaw4597"},{"key":"e_1_2_12_2_31_1","doi-asserted-by":"publisher","DOI":"10.1172\/jci.insight.121689"},{"key":"e_1_2_12_2_32_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2015.06.002"},{"key":"e_1_2_12_2_33_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cmet.2014.12.001"},{"key":"e_1_2_12_2_34_1","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCULATIONAHA.118.037968"},{"key":"e_1_2_12_2_35_1","doi-asserted-by":"publisher","DOI":"10.1172\/jci.insight.134063"},{"key":"e_1_2_12_2_36_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature22082"},{"key":"e_1_2_12_2_37_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2015.06.017"},{"key":"e_1_2_12_2_38_1","doi-asserted-by":"publisher","DOI":"10.3389\/fphys.2020.510600"},{"key":"e_1_2_12_2_39_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M403607200"},{"key":"e_1_2_12_2_40_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0091-679X(06)80001-4"},{"key":"e_1_2_12_2_41_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0908099107"},{"key":"e_1_2_12_2_42_1","doi-asserted-by":"publisher","DOI":"10.1038\/ncb2868"},{"key":"e_1_2_12_2_43_1","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.adp7444"},{"key":"e_1_2_12_2_44_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2023.112735"},{"key":"e_1_2_12_2_45_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.yjmcc.2022.11.003"},{"key":"e_1_2_12_2_46_1","doi-asserted-by":"publisher","DOI":"10.1038\/srep34174"},{"key":"e_1_2_12_2_47_1","doi-asserted-by":"publisher","DOI":"10.1085\/jgp.201311096"},{"key":"e_1_2_12_2_48_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ceca.2024.102907"},{"key":"e_1_2_12_2_49_1","doi-asserted-by":"crossref","unstructured":"Vetralla 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