{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,3]],"date-time":"2026-10-03T12:15:43Z","timestamp":1791029743824,"version":"4.1.0"},"reference-count":31,"publisher":"National Academy of Sciences","issue":"51","content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2010,12,21]]},"abstract":"<jats:p>\n                    The cytosolic free Ca\n                    <jats:sup>2+<\/jats:sup>\n                    transients elicited by muscle fiber excitation are well characterized, but little is known about the free [Ca\n                    <jats:sup>2+<\/jats:sup>\n                    ] dynamics within the sarcoplasmic reticulum (SR). A targetable ratiometric FRET-based calcium indicator (D1ER Cameleon) allowed us to investigate SR Ca\n                    <jats:sup>2+<\/jats:sup>\n                    dynamics and analyze the impact of calsequestrin (CSQ) on SR [Ca\n                    <jats:sup>2+<\/jats:sup>\n                    ] in enzymatically dissociated flexor digitorum brevis muscle fibers from WT and CSQ-KO mice lacking isoform 1 (CSQ-KO) or both isoforms [CSQ-double KO (DKO)]. At rest, free SR [Ca\n                    <jats:sup>2+<\/jats:sup>\n                    ] did not differ between WT, CSQ-KO, and CSQ-DKO fibers. During sustained contractions, changes were rather small in WT, reflecting powerful buffering of CSQ, whereas in CSQ-KO fibers, significant drops in SR [Ca\n                    <jats:sup>2+<\/jats:sup>\n                    ] occurred. Their amplitude increased with stimulation frequency between 1 and 60 Hz. At 60 Hz, the SR became virtually depleted of Ca\n                    <jats:sup>2+<\/jats:sup>\n                    , both in CSQ-KO and CSQ-DKO fibers. In CSQ-KO fibers, cytosolic free calcium detected with Fura-2 declined during repetitive stimulation, indicating that SR calcium content was insufficient for sustained contractile activity. SR Ca\n                    <jats:sup>2+<\/jats:sup>\n                    reuptake during and after stimulation trains appeared to be governed by three temporally distinct processes with rate constants of 50, 1\u20135, and 0.3 s\n                    <jats:sup>\u22121<\/jats:sup>\n                    (at 26 \u00b0C), reflecting activity of the SR Ca\n                    <jats:sup>2+<\/jats:sup>\n                    pump and interplay of luminal and cytosolic Ca\n                    <jats:sup>2+<\/jats:sup>\n                    buffers and pointing to store-operated calcium entry (SOCE). SOCE might play an essential role during muscle contractures responsible for the malignant hyperthermia-like syndrome in mice lacking CSQ.\n                  <\/jats:p>","DOI":"10.1073\/pnas.1009168108","type":"journal-article","created":{"date-parts":[[2010,12,6]],"date-time":"2010-12-06T23:17:29Z","timestamp":1291677449000},"page":"22326-22331","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":89,"title":["Massive alterations of sarcoplasmic reticulum free calcium in skeletal muscle fibers lacking calsequestrin revealed by a genetically encoded probe"],"prefix":"10.1073","volume":"107","author":[{"given":"M.","family":"Canato","sequence":"first","affiliation":[{"name":"Departments of aHuman Anatomy and Physiology and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M.","family":"Scorzeto","sequence":"additional","affiliation":[{"name":"Departments of aHuman Anatomy and Physiology and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M.","family":"Giacomello","sequence":"additional","affiliation":[{"name":"Experimental Veterinary Sciences, University of Padua, 35121 Padua, Italy;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"F.","family":"Protasi","sequence":"additional","affiliation":[{"name":"CeSI, Department of Basic and Applied Medical Sciences, University G. d'Annunzio, I-66013 Chieti, Italy;"},{"name":"IIM-Interuniversity Institute of Myology and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"C.","family":"Reggiani","sequence":"additional","affiliation":[{"name":"Departments of aHuman Anatomy and Physiology and"},{"name":"IIM-Interuniversity Institute of Myology and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"G. 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