{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,31]],"date-time":"2026-10-31T16:43:19Z","timestamp":1793464999548,"version":"4.5.0"},"reference-count":45,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2024,2,21]],"date-time":"2024-02-21T00:00:00Z","timestamp":1708473600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Neurosci."],"abstract":"<jats:p>Disease-associated variants of <jats:italic>TUBA4A<\/jats:italic> (alpha-tubulin 4A) have recently been identified in familial ALS. Interestingly, a downregulation of TUBA4A protein expression was observed in familial as well as sporadic ALS brain tissue. To investigate whether a decreased TUBA4A expression could be a driving factor in ALS pathogenesis, we assessed whether <jats:italic>TUBA4A<\/jats:italic> knockdown in zebrafish could recapitulate an ALS-like phenotype. For this, we injected an antisense oligonucleotide morpholino in zebrafish embryos targeting the zebrafish <jats:italic>TUBA4A<\/jats:italic> orthologue. An antibody against synaptic vesicle 2 was used to visualize motor axons in the spinal cord, allowing the analysis of embryonic ventral root projections. Motor behavior was assessed using the touch-evoked escape response. In <jats:italic>post-mortem<\/jats:italic> ALS motor cortex, we observed reduced TUBA4A levels. The knockdown of the zebrafish <jats:italic>TUBA4A<\/jats:italic> orthologue induced a motor axonopathy and a significantly disturbed motor behavior. Both phenotypes were dose-dependent and could be rescued by the addition of human wild-type <jats:italic>TUBA4A<\/jats:italic> mRNA. Thus, <jats:italic>TUBA4A<\/jats:italic> downregulation as observed in ALS <jats:italic>post-mortem<\/jats:italic> motor cortex could be modeled in zebrafish and induced a motor axonopathy and motor behavior defects reflecting a motor neuron disease phenotype, as previously described in embryonic zebrafish models of ALS. The rescue with human wild-type <jats:italic>TUBA4A<\/jats:italic> mRNA suggests functional conservation and strengthens the causal relation between TUBA4A protein levels and phenotype severity. Furthermore, the loss of TUBA4A induces significant changes in post-translational modifications of tubulin, such as acetylation, detyrosination and polyglutamylation. Our data unveil an important role for <jats:italic>TUBA4A<\/jats:italic> in ALS pathogenesis, and extend the relevance of <jats:italic>TUBA4A<\/jats:italic> to the majority of ALS patients, in addition to cases bearing <jats:italic>TUBA4A<\/jats:italic> mutations.<\/jats:p>","DOI":"10.3389\/fncel.2024.1340240","type":"journal-article","created":{"date-parts":[[2024,2,21]],"date-time":"2024-02-21T04:43:31Z","timestamp":1708490611000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":6,"title":["TUBA4A downregulation as observed in ALS post-mortem motor cortex causes ALS-related abnormalities in zebrafish"],"prefix":"10.3389","volume":"18","author":[{"given":"Evelien","family":"Van Schoor","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dufie","family":"Strubbe","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elke","family":"Braems","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jochen","family":"Weishaupt","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Albert C.","family":"Ludolph","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Philip","family":"Van Damme","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dietmar Rudolf","family":"Thal","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Val\u00e9rie","family":"Bercier","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ludo","family":"Van Den Bosch","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2024,2,21]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"617","DOI":"10.1038\/nrneurol.2013.203","article-title":"The epidemiology of ALS: a conspiracy of genes, environment and time.","volume":"9","author":"Al-Chalabi","year":"2013","journal-title":"Nat. Rev. Neurol."},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1186\/s13024-019-0327-3","article-title":"Dynactin1 depletion leads to neuromuscular synapse instability and functional abnormalities.","volume":"14","author":"Bercier","year":"2019","journal-title":"Mol. Neurodegeneration"},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.1016\/j.neulet.2021.135656","article-title":"Tubulin polyglutamylation, a regulator of microtubule functions, can cause neurodegeneration.","volume":"746","author":"Bodakuntla","year":"2021","journal-title":"Neurosci. Lett."},{"key":"B4","doi-asserted-by":"publisher","first-page":"604","DOI":"10.1016\/j.drudis.2020.11.033","article-title":"Microtubule-targeting agents and neurodegeneration.","volume":"26","author":"Boiarska","year":"2021","journal-title":"Drug Discov. Today"},{"key":"B5","doi-asserted-by":"publisher","first-page":"389","DOI":"10.1007\/s00401-006-0127-z","article-title":"Staging of Alzheimer disease-associated neurofibrillary pathology using paraffin sections and immunocytochemistry.","volume":"112","author":"Braak","year":"2006","journal-title":"Acta Neuropathol."},{"key":"B6","doi-asserted-by":"publisher","first-page":"819","DOI":"10.1016\/j.ajhg.2010.03.019","article-title":"Tuba8 is expressed at low levels in the developing mouse and human brain.","volume":"86","author":"Braun","year":"2010","journal-title":"Am. J. Hum. Genet."},{"key":"B7","doi-asserted-by":"publisher","first-page":"58","DOI":"10.1016\/j.mcn.2017.03.002","article-title":"Tubulins and brain development \u2013 The origins of functional specification.","volume":"84","author":"Breuss","year":"2017","journal-title":"Mol. Cell. Neurosci."},{"key":"B8","doi-asserted-by":"publisher","first-page":"293","DOI":"10.1080\/146608200300079536","article-title":"El escorial revisited: revised criteria for the diagnosis of amyotrophic lateral sclerosis.","volume":"1","author":"Brooks","year":"2000","journal-title":"Amyotroph. Lateral Scler."},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.1016\/j.neulet.2021.135900","article-title":"Microtubule acetylation: a reading key to neural physiology and degeneration.","volume":"755","author":"Cappelletti","year":"2021","journal-title":"Neurosci. Lett."},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.3389\/fncel.2020.594975","article-title":"The neglected genes of ALS: cytoskeletal dynamics impact synaptic degeneration in ALS.","volume":"14","author":"Castellanos-Montiel","year":"2020","journal-title":"Front. Cell. Neurosci."},{"key":"B11","doi-asserted-by":"publisher","first-page":"521","DOI":"10.1002\/cm.21290","article-title":"The emerging role of the tubulin code: from the tubulin molecule to neuronal function and disease.","volume":"73","author":"Chakraborti","year":"2016","journal-title":"Cytoskeleton"},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.3389\/fncel.2016.00204","article-title":"A case for microtubule vulnerability in amyotrophic lateral sclerosis: altered dynamics during disease.","volume":"10","author":"Clark","year":"2016","journal-title":"Front. Cell. Neurosci."},{"key":"B13","doi-asserted-by":"publisher","first-page":"497","DOI":"10.1016\/j.clinph.2007.09.143","article-title":"Electrodiagnostic criteria for diagnosis of ALS.","volume":"119","author":"de Carvalho","year":"2008","journal-title":"Clin. Neurophysiol."},{"key":"B14","doi-asserted-by":"publisher","first-page":"53","DOI":"10.1080\/17482960802521126","article-title":"Awaji diagnostic algorithm increases sensitivity of El Escorial criteria for ALS diagnosis.","volume":"10","author":"de Carvalho","year":"2009","journal-title":"Amyotroph. Lateral Scler."},{"key":"B15","doi-asserted-by":"publisher","first-page":"232","DOI":"10.1016\/j.expneurol.2003.10.004","article-title":"Amyotrophic lateral sclerosis is a distal axonopathy: evidence in mice and man.","volume":"185","author":"Fischer","year":"2004","journal-title":"Exp. Neurol."},{"key":"B16","doi-asserted-by":"publisher","first-page":"133","DOI":"10.1016\/j.semcdb.2019.07.010","article-title":"Axonal transport defects and neurodegeneration: molecular mechanisms and therapeutic implications.","volume":"99","author":"Guo","year":"2020","journal-title":"Semin. Cell Dev. Biol."},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-022-31776-5","article-title":"Disruption of tubulin-alpha4a polyglutamylation prevents aggregation of hyper-phosphorylated tau and microglia activation in mice.","volume":"13","author":"Hausrat","year":"2022","journal-title":"Nat. Commun."},{"key":"B18","doi-asserted-by":"publisher","first-page":"4301","DOI":"10.1007\/s00018-018-2873-1","article-title":"Dysregulation of a novel miR-1825\/TBCB\/TUBA4A pathway in sporadic and familial ALS.","volume":"75","author":"Helferich","year":"2018","journal-title":"Cell. Mol. Life Sci."},{"key":"B19","doi-asserted-by":"publisher","first-page":"307","DOI":"10.1038\/s41580-020-0214-3","article-title":"The tubulin code and its role in controlling microtubule properties and functions.","volume":"21","author":"Janke","year":"2020","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"B20","doi-asserted-by":"publisher","first-page":"236","DOI":"10.1002\/ana.20379","article-title":"Gene expression profile of spinal motor neurons in sporadic amyotrophic lateral sclerosis.","volume":"57","author":"Jiang","year":"2005","journal-title":"Ann. Neurol."},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.1371\/JOURNAL.PGEN.1002214","article-title":"FUS and TARDBP but not SOD1 interact in genetic models of amyotrophic lateral sclerosis.","volume":"7","author":"Kabashi","year":"2011","journal-title":"PLoS Genet."},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0013368","article-title":"Progranulin is neurotrophic in vivo and protects against a mutant TDP-43 induced axonopathy.","volume":"5","author":"Laird","year":"2010","journal-title":"PLoS one"},{"key":"B23","doi-asserted-by":"publisher","first-page":"214","DOI":"10.1002\/cm.20436","article-title":"Tumoral and tissue-specific expression of the major human beta-tubulin isotypes.","volume":"67","author":"Leandro-Garc\u00eda","year":"2010","journal-title":"Cytoskeleton"},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.15252\/embj.2018100440","article-title":"Excessive tubulin polyglutamylation causes neurodegeneration and perturbs neuronal transport.","volume":"37","author":"Magiera","year":"2018","journal-title":"EMBO J."},{"key":"B25","doi-asserted-by":"publisher","first-page":"209","DOI":"10.1016\/j.neuro.2019.09.015","article-title":"Influence of selenium on the emergence of neuro tubule defects in a neuron-like cell line and its implications for amyotrophic lateral sclerosis.","volume":"75","author":"Maraldi","year":"2019","journal-title":"Neurotoxicology"},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.1212\/NXG.0000000000000596","article-title":"Novel TUBA4A variant associated with familial frontotemporal dementia.","volume":"7","author":"Mol","year":"2021","journal-title":"Neurol. Genet."},{"key":"B27","doi-asserted-by":"publisher","first-page":"253","DOI":"10.1002\/dneu.22774","article-title":"Tubulin post-translational modifications control neuronal development and functions.","volume":"81","author":"Moutin","year":"2021","journal-title":"Dev. Neurobiol."},{"key":"B28","doi-asserted-by":"publisher","first-page":"863","DOI":"10.1007\/s00401-017-1708-8","article-title":"Motor neuron vulnerability and resistance in amyotrophic lateral sclerosis.","volume":"133","author":"Nijssen","year":"2017","journal-title":"Acta Neuropathol."},{"key":"B29","doi-asserted-by":"publisher","first-page":"891","DOI":"10.1111\/nan.12712","article-title":"An autopsy case of pure nigropathy with TUBA4A nonsense mutation.","volume":"47","author":"Okada","year":"2021","journal-title":"Neuropathol. Appl. Neurobiol."},{"key":"B30","doi-asserted-by":"publisher","first-page":"177.e9","DOI":"10.1016\/j.neurobiolaging.2016.12.008","article-title":"Investigating the role of ALS genes CHCHD10 and TUBA4A in Belgian FTD-ALS spectrum patients.","volume":"51","author":"Perrone","year":"2017","journal-title":"Neurobiol. Aging"},{"key":"B32","doi-asserted-by":"publisher","first-page":"46","DOI":"10.1016\/j.semcdb.2021.12.006","article-title":"The detyrosination\/re-tyrosination cycle of tubulin and its role and dysfunction in neurons and cardiomyocytes.","volume":"137","author":"Sanyal","year":"2021","journal-title":"Sem. Cell Dev. Biol."},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.3390\/ijms21197354","article-title":"Microtubule dysfunction: a common feature of neurodegenerative diseases.","volume":"21","author":"Sferra","year":"2020","journal-title":"Int. J. Mol. Sci."},{"key":"B34","doi-asserted-by":"publisher","first-page":"324","DOI":"10.1016\/j.neuron.2014.09.027","article-title":"Exome-wide rare variant analysis identifies TUBA4A mutations associated with familial ALS.","volume":"84","author":"Smith","year":"2014","journal-title":"Neuron"},{"key":"B35","doi-asserted-by":"publisher","first-page":"2655","DOI":"10.1523\/JNEUROSCI.23-07-02655.2003","article-title":"Visualization of microtubule growth in cultured neurons via the use of EB3-GFP (end-binding Protein 3-green fluorescent protein).","volume":"23","author":"Stepanova","year":"2003","journal-title":"J. Neurosci."},{"key":"B36","doi-asserted-by":"publisher","first-page":"2213","DOI":"10.1016\/j.stemcr.2021.03.029","article-title":"Human motor units in microfluidic devices are impaired by FUS mutations and improved by HDAC6 inhibition.","volume":"16","author":"Stoklund Dittlau","year":"2021","journal-title":"Stem. Cell Rep."},{"key":"B37","doi-asserted-by":"publisher","first-page":"427","DOI":"10.1007\/s00401-017-1796-5","article-title":"A zebrafish model for C9orf72 ALS reveals RNA toxicity as a pathogenic mechanism.","volume":"135","author":"Swinnen","year":"2018","journal-title":"Acta Neuropathol."},{"key":"B38","doi-asserted-by":"publisher","first-page":"197","DOI":"10.1038\/nature20413","article-title":"Decoding ALS: from genes to mechanism.","volume":"539","author":"Taylor","year":"2016","journal-title":"Nature"},{"key":"B31","doi-asserted-by":"publisher","first-page":"733","DOI":"10.1093\/jnen\/59.8.733","article-title":"Sequence of A\u03b2-protein deposition in the human medial temporal lobe.","volume":"59","author":"Thal","year":"2000","journal-title":"J. Neuropathol. Exp. Neurol."},{"key":"B39","author":"Thisse","year":"2004","journal-title":"Fast Release Clones: a High Throughput Expression Analysis., ZFIN Direct Data Submission."},{"key":"B40","doi-asserted-by":"publisher","first-page":"286","DOI":"10.1016\/j.gde.2011.01.003","article-title":"Phenotypic spectrum of the tubulin-related disorders and functional implications of disease-causing mutations.","volume":"21","author":"Tischfield","year":"2011","journal-title":"Curr. Opin. Genet. Dev."},{"key":"B41","doi-asserted-by":"publisher","first-page":"1418","DOI":"10.1038\/nm.2901","article-title":"EPHA4 is a disease modifier of amyotrophic lateral sclerosis in animal models and in humans.","volume":"18","author":"Van Hoecke","year":"2012","journal-title":"Nat. Med."},{"key":"B42","doi-asserted-by":"publisher","DOI":"10.3390\/biom12030440","article-title":"Frontotemporal lobar degeneration case with an N-Terminal TUBA4A mutation exhibits reduced TUBA4A levels in the brain and TDP-43 pathology.","volume":"12","author":"Van Schoor","year":"2022","journal-title":"Biomolecules"},{"key":"B43","doi-asserted-by":"publisher","first-page":"2981","DOI":"10.1021\/acschembio.6b00507","article-title":"Molecular determinants of tubulin\u2019s C-terminal tail conformational ensemble.","volume":"11","author":"Wall","year":"2016","journal-title":"ACS Chem. Biol."},{"key":"B44","doi-asserted-by":"publisher","DOI":"10.1101\/2022.02.17.480930","article-title":"Asymmetric requirement for \u03b1-tubulin over \u03b2-tubulin.","volume":"222","author":"Wethekam","year":"2022","journal-title":"J. Cell Biol."},{"key":"B45","doi-asserted-by":"publisher","first-page":"546","DOI":"10.1002\/dvg.20766","article-title":"In vivo labeling of zebrafish motor neurons using an mnx1 enhancer and Gal4\/UAS.","volume":"49","author":"Zelenchuk","year":"2011","journal-title":"Genesis"}],"container-title":["Frontiers in Cellular Neuroscience"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fncel.2024.1340240\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,2,23]],"date-time":"2024-02-23T14:13:34Z","timestamp":1708697614000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fncel.2024.1340240\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,2,21]]},"references-count":45,"alternative-id":["10.3389\/fncel.2024.1340240"],"URL":"https:\/\/doi.org\/10.3389\/fncel.2024.1340240","relation":{},"ISSN":["1662-5102"],"issn-type":[{"value":"1662-5102","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,2,21]]},"article-number":"1340240"}}