{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,27]],"date-time":"2026-03-27T16:06:53Z","timestamp":1774627613380,"version":"3.50.1"},"reference-count":53,"publisher":"Oxford University Press (OUP)","issue":"8","license":[{"start":{"date-parts":[[2021,3,9]],"date-time":"2021-03-09T00:00:00Z","timestamp":1615248000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004837","name":"Spanish Ministry of Science and Innovation","doi-asserted-by":"publisher","award":["PID2019-110272R"],"award-info":[{"award-number":["PID2019-110272R"]}],"id":[{"id":"10.13039\/501100004837","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004837","name":"Spanish Ministry of Science and Innovation","doi-asserted-by":"publisher","award":["BES-2014-069808"],"award-info":[{"award-number":["BES-2014-069808"]}],"id":[{"id":"10.13039\/501100004837","id-type":"DOI","asserted-by":"publisher"}]},{"name":"SMA-Europe"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2021,5,17]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>Spinal muscular atrophy (SMA) is an autosomal recessive degenerative motor neuron disease characterized by symmetrical muscle weakness and atrophy of limb and trunk muscles being the most severe genetic disease in children. In SMA mouse models, motor nerve terminals display neurotransmitter release reduction, endocytosis decrease and mitochondria alterations. The relationship between these changes is, however, not well understood. In the present study, we investigated whether the endocytosis impairment could be related to the functional alteration of the presynaptic mitochondria during action potential (AP) firing. To this aim, we generated a Synaptophysin-pHluorin (SypHy) transgenic mouse, crossed it with Taiwanese SMA mice, and recorded exo- and endocytosis and mitochondria Ca2+ signaling in real-time at ex vivo motor nerve terminals of Taiwanese-SypHy mice. The experiments were performed at the beginning of the motor symptoms to get an integrated view of the nerve terminal\u2019s functional state before degeneration. Our electrophysiological and live imaging results demonstrated that the mitochondria\u2019s capacity to increase matrix-free Ca2+ in SMA mice was significantly limited during nerve AP firing, except when the rate of Ca2+ entry to the cytosol was considerably reduced. These results indicate that both the mitochondrial Ca2+ signaling alterations and the secretion machinery defects are significant players in the dysfunction of the presynaptic terminal in SMA.<\/jats:p>","DOI":"10.1093\/hmg\/ddab065","type":"journal-article","created":{"date-parts":[[2021,2,25]],"date-time":"2021-02-25T20:12:40Z","timestamp":1614283960000},"page":"629-643","source":"Crossref","is-referenced-by-count":15,"title":["Calcium is reduced in presynaptic mitochondria of motor nerve terminals during neurotransmission in SMA mice"],"prefix":"10.1093","volume":"30","author":[{"given":"Mario","family":"Lopez-Manzaneda","sequence":"first","affiliation":[{"name":"Department of Medical Physiology and Biophysics, School of Medicine, University of Seville, 41009 Seville, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Julio","family":"Franco-Espin","sequence":"additional","affiliation":[{"name":"Department of Medical Physiology and Biophysics, School of Medicine, University of Seville, 41009 Seville, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rocio","family":"Tejero","sequence":"additional","affiliation":[{"name":"Department of Medical Physiology and Biophysics, School of Medicine, University of Seville, 41009 Seville, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Raquel","family":"Cano","sequence":"additional","affiliation":[{"name":"Department of Medical Physiology and Biophysics, School of Medicine, University of Seville, 41009 Seville, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4027-4044","authenticated-orcid":false,"given":"Lucia","family":"Tabares","sequence":"additional","affiliation":[{"name":"Department of Medical Physiology and Biophysics, School of Medicine, University of Seville, 41009 Seville, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2021,3,9]]},"reference":[{"key":"2021051711184667100_ref1","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1038\/ejhg.2011.134","article-title":"Pan-ethnic carrier screening and prenatal diagnosis for spinal muscular atrophy: clinical laboratory analysis of &gt;72 400 specimens","volume":"20","author":"Sugarman","year":"2012","journal-title":"Eur. 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Cell Res."},{"key":"2021051711184667100_ref21","doi-asserted-by":"crossref","first-page":"436","DOI":"10.1073\/pnas.0908099107","article-title":"NCLX is an essential component of mitochondrial Na+\/Ca2+ exchange","volume":"107","author":"Palty","year":"2010","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"key":"2021051711184667100_ref22","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.cophys.2018.02.009","article-title":"Maturation and heterogeneity of vertebrate motor synapses","volume":"4","author":"Tejero","year":"2018","journal-title":"Curr. Opin. Physio."},{"key":"2021051711184667100_ref23","doi-asserted-by":"crossref","first-page":"1003","DOI":"10.1016\/S0896-6273(00)81046-6","article-title":"SV2A and SV2B function as redundant Ca2+ regulators in neurotransmitter release","volume":"24","author":"Janz","year":"1999","journal-title":"Neuron"},{"key":"2021051711184667100_ref24","doi-asserted-by":"crossref","first-page":"691","DOI":"10.1038\/35087000","article-title":"SV2 modulates the size of the readily releasable pool of secretory vesicles","volume":"3","author":"Xu","year":"2001","journal-title":"Nat. Cell Biol."},{"key":"2021051711184667100_ref25","doi-asserted-by":"crossref","first-page":"5569","DOI":"10.1523\/JNEUROSCI.4781-09.2010","article-title":"Cotrafficking of SV2 and synaptotagmin at the synapse","volume":"30","author":"Yao","year":"2010","journal-title":"J. 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Neurosci."},{"key":"2021051711184667100_ref29","doi-asserted-by":"crossref","first-page":"6467","DOI":"10.1523\/JNEUROSCI.21-17-06467.2001","article-title":"ATP is required at an early step in compensatory endocytosis in synaptic terminals","volume":"21","author":"Heidelberger","year":"2001","journal-title":"J. Neurosci."},{"key":"2021051711184667100_ref30","doi-asserted-by":"crossref","first-page":"825","DOI":"10.1016\/j.cell.2013.12.042","article-title":"Activity-driven local ATP synthesis is required for synaptic function","volume":"156","author":"Rangaraju","year":"2014","journal-title":"Cell"},{"key":"2021051711184667100_ref31","doi-asserted-by":"crossref","first-page":"491","DOI":"10.1152\/jn.00012.2003","article-title":"Inhibition of mitochondrial Ca2+ uptake affects phasic release from motor terminals differently depending on external [Ca2+]","volume":"90","author":"Talbot","year":"2003","journal-title":"J. Neurophysiol."},{"key":"2021051711184667100_ref32","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1038\/35000001","article-title":"Chromaffin-cell stimulation triggers fast millimolar mitochondrial Ca2+ transients that modulate secretion","volume":"2","author":"Montero","year":"2000","journal-title":"Nat. Cell Biol."},{"key":"2021051711184667100_ref33","doi-asserted-by":"crossref","first-page":"833","DOI":"10.1083\/jcb.136.4.833","article-title":"Mitochondrial participation in the intracellular Ca2+ network","volume":"136","author":"Babcock","year":"1997","journal-title":"J. Cell Biol."},{"key":"2021051711184667100_ref34","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1111\/j.1469-7793.1998.059bo.x","article-title":"Evidence that mitochondria buffer physiological Ca2+ loads in lizard motor nerve terminals","volume":"509","author":"David","year":"1998","journal-title":"J. Physiol."},{"key":"2021051711184667100_ref35","doi-asserted-by":"crossref","first-page":"7290","DOI":"10.1523\/JNEUROSCI.20-19-07290.2000","article-title":"Stimulation-evoked increases in cytosolic [Ca2+] in mouse motor nerve terminals are limited by mitochondrial uptake and are temperature-dependent","volume":"20","author":"David","year":"2000","journal-title":"J. Neurosci."},{"key":"2021051711184667100_ref36","doi-asserted-by":"crossref","first-page":"197","DOI":"10.1016\/S0143-4160(02)00229-4","article-title":"Quantitative estimate of mitochondrial [Ca2+] in stimulated motor nerve terminals","volume":"33","author":"David","year":"2003","journal-title":"Cell Calcium"},{"key":"2021051711184667100_ref37","doi-asserted-by":"crossref","first-page":"3440","DOI":"10.1016\/S0021-9258(19)83641-4","article-title":"A new generation of Ca2+ indicators with greatly improved fluorescence properties","volume":"260","author":"Grynkiewicz","year":"1985","journal-title":"J. Biol. Chem."},{"key":"2021051711184667100_ref38","doi-asserted-by":"crossref","first-page":"7495","DOI":"10.1523\/JNEUROSCI.19-17-07495.1999","article-title":"Mitochondrial clearance of cytosolic Ca2+ in stimulated lizard motor nerve terminals proceeds without progressive elevation of mitochondrial matrix [Ca2+]","volume":"19","author":"David","year":"1999","journal-title":"J. Neurosci."},{"key":"2021051711184667100_ref39","doi-asserted-by":"crossref","first-page":"19062","DOI":"10.1074\/jbc.M212661200","article-title":"The relationship between free and total calcium concentrations in the matrix of liver and brain mitochondria","volume":"278","author":"Chalmers","year":"2003","journal-title":"J. Biol. Chem."},{"key":"2021051711184667100_ref40","doi-asserted-by":"crossref","first-page":"376","DOI":"10.1016\/j.celrep.2015.08.079","article-title":"PKA phosphorylation of NCLX reverses mitochondrial calcium overload and depolarization, promoting survival of PINK1- deficient dopaminergic neurons","volume":"13","author":"Kostic","year":"2015","journal-title":"Cell Rep."},{"key":"2021051711184667100_ref41","doi-asserted-by":"crossref","first-page":"333","DOI":"10.1111\/j.1432-1033.1982.tb06875.x","article-title":"The role of phosphate in the regulation of the independent calcium-efflux pathway of liver mitochondria","volume":"127","author":"Zoccarato","year":"1982","journal-title":"Eur. J. Biochem."},{"key":"2021051711184667100_ref42","doi-asserted-by":"crossref","first-page":"2007","DOI":"10.1073\/pnas.0810934106","article-title":"The \u03a8 m depolarization that accompanies mitochondrial Ca2+ uptake is greater in mutant SOD1 than in wild-type mouse motor terminals","volume":"106","author":"Nguyen","year":"2009","journal-title":"Proc. Natl. Acad. Sci."},{"key":"2021051711184667100_ref43","doi-asserted-by":"crossref","first-page":"100826","DOI":"10.1016\/j.isci.2020.100826","article-title":"R-roscovitine improves motoneuron function in mouse models for spinal muscular atrophy","volume":"23","author":"Tejero","year":"2020","journal-title":"iScience"},{"key":"2021051711184667100_ref44","doi-asserted-by":"crossref","first-page":"425","DOI":"10.1113\/jphysiol.2002.035196","article-title":"Mitochondrial Ca2+ uptake prevents desynchronization of quantal release and minimizes depletion during repetitive stimulation of mouse motor nerve terminals","volume":"548","author":"David","year":"2003","journal-title":"J. Physiol."},{"key":"2021051711184667100_ref45","doi-asserted-by":"crossref","first-page":"663","DOI":"10.1113\/jphysiol.2006.110841","article-title":"Extrusion of Ca2+ from mouse motor terminal mitochondria via a Na+-Ca2+ exchanger increases post-tetanic evoked release","volume":"574","author":"Garc\u00eda-Chac\u00f3n","year":"2006","journal-title":"J. Physiol."},{"key":"2021051711184667100_ref46","doi-asserted-by":"crossref","first-page":"1349","DOI":"10.1111\/j.1471-4159.2006.03619.x","article-title":"Neural mitochondrial Ca2+ capacity impairment precedes the onset of motor symptoms in G93A Cu\/Zn-superoxide dismutase mutant mice","volume":"96","author":"Damiano","year":"2006","journal-title":"J. Neurochem."},{"key":"2021051711184667100_ref47","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1083\/jcb.200703187","article-title":"Defective Ca2+ channel clustering in axon terminals disturbs excitability in motoneurons in spinal muscular atrophy","volume":"179","author":"Jablonka","year":"2007","journal-title":"J. Cell Biol."},{"key":"2021051711184667100_ref48","doi-asserted-by":"crossref","first-page":"277","DOI":"10.1023\/B:JOBB.0000041753.52832.f3","article-title":"The integration of mitochondrial calcium transport and storage","volume":"36","author":"Nicholls","year":"2004","journal-title":"J. Bioenerg. Biomembr."},{"key":"2021051711184667100_ref49","doi-asserted-by":"crossref","first-page":"391","DOI":"10.1152\/physrev.1990.70.2.391","article-title":"Role of calcium ions in regulation of mammalian intramitochondrial metabolism","volume":"70","author":"McCormack","year":"1990","journal-title":"Physiol. Rev."},{"key":"2021051711184667100_ref50","doi-asserted-by":"crossref","first-page":"13807","DOI":"10.1073\/pnas.96.24.13807","article-title":"Regulation of mitochondrial ATP synthesis by calcium: evidence for a long-term metabolic priming","volume":"96","author":"Jouaville","year":"1999","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"key":"2021051711184667100_ref51","doi-asserted-by":"crossref","first-page":"66","DOI":"10.1038\/71709","article-title":"A mouse model for spinal muscular atrophy","volume":"24","author":"Hsieh-Li","year":"2000","journal-title":"Nat. Genet."},{"key":"2021051711184667100_ref52","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1113\/jphysiol.1981.sp013586","article-title":"Non-linear summation of endplate potentials in the frog and mouse","volume":"311","author":"McLachlan","year":"1981","journal-title":"J. Physiol."},{"key":"2021051711184667100_ref53","doi-asserted-by":"crossref","first-page":"2000","DOI":"10.1523\/JNEUROSCI.4663-10.2011","article-title":"Active zones and the readily releasable pool of synaptic vesicles at the neuromuscular junction of the mouse","volume":"31","author":"Ruiz","year":"2011","journal-title":"J. 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