{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,2]],"date-time":"2025-11-02T09:53:42Z","timestamp":1762077222430,"version":"build-2065373602"},"reference-count":32,"publisher":"MDPI AG","issue":"22","license":[{"start":{"date-parts":[[2022,11,18]],"date-time":"2022-11-18T00:00:00Z","timestamp":1668729600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003554","name":"Lundbeck Foundation","doi-asserted-by":"publisher","award":["R192-2015-895","R223-2015-4222","R248-2016-2518","R155-2013-14113"],"award-info":[{"award-number":["R192-2015-895","R223-2015-4222","R248-2016-2518","R155-2013-14113"]}],"id":[{"id":"10.13039\/501100003554","id-type":"DOI","asserted-by":"publisher"}]},{"name":"EC Framework 7 Marie Curie Fellowship, Training Network Grant (NEURASYNC)","award":["R192-2015-895","R223-2015-4222","R248-2016-2518","R155-2013-14113"],"award-info":[{"award-number":["R192-2015-895","R223-2015-4222","R248-2016-2518","R155-2013-14113"]}]},{"name":"EU FP7 MEFOPA","award":["R192-2015-895","R223-2015-4222","R248-2016-2518","R155-2013-14113"],"award-info":[{"award-number":["R192-2015-895","R223-2015-4222","R248-2016-2518","R155-2013-14113"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Overexpression of \u03b1-synuclein with tyrosine mutated to phenylalanine at position 125 leads to a severe phenotype with motor impairment and neuropathology in Drosophila. Here, we hypothesized that tyrosine mutations would similarly lead to impaired motor performance with neuropathology in a rodent model. In transgenic mice (ASO), tyrosines at positions 125, 133, and 136 in human \u03b1-synuclein were mutated to phenylalanine and cloned into a Thy1.2 expression vector, which was used to create transgenic mouse lines on a mixed genetic background TgN(Thy-1-SNCA-YF)4Emfu (YF). The YF mice had a decreased lifespan and displayed a dramatic motor phenotype with paralysis of both hind- and forelegs. Post-translational modification of \u03b1-synuclein due to phosphorylation of serine 129 is often seen in inclusions in the brains of patients with \u03b1-synucleinopathies. We observed a slight but significant increase in phosphorylation of serine 129 in the cytosol in YF mice compared to age-matched human \u03b1-synuclein transgenic mice (ASO). Conversely, significantly decreased phosphorylation of serine 129 was seen in synaptosomes of YF mice that also contained higher amounts of soluble oligomers. YF mice deposited full-length \u03b1-synuclein aggregates in neurons widespread in the CNS with the main occurrence in the forebrain structures of the cerebral cortex, the basal ganglia, and limbic structures. Full-length \u03b1-synuclein labeling was also prominent in many nuclear regions of the brain stem, deep cerebellar nuclei, and cerebellar cortex. The study shows that the substitution of tyrosines to phenylalanine in \u03b1-synuclein at positions 125, 133, and 136 leads to severe toxicity in vivo. An insignificant change upon tyrosine substitution suggests that the phosphorylation of serine 129 is not the cause of the toxicity.<\/jats:p>","DOI":"10.3390\/cells11223673","type":"journal-article","created":{"date-parts":[[2022,11,21]],"date-time":"2022-11-21T03:11:21Z","timestamp":1669000281000},"page":"3673","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Mutation of Tyrosine Sites in the Human Alpha-Synuclein Gene Induces Neurotoxicity in Transgenic Mice with Soluble Alpha-Synuclein Oligomer Formation"],"prefix":"10.3390","volume":"11","author":[{"given":"Louise Berkhoudt","family":"Lassen","sequence":"first","affiliation":[{"name":"DANDRITE, Department of Biomedicine, Aarhus University, 8000 Aarhus, Denmark"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maj Schneider","family":"Thomsen","sequence":"additional","affiliation":[{"name":"Neurobiology Research and Drug Delivery, Department of Health Science and Technology, Aalborg University, 9220 Aalborg, Denmark"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Elisa","family":"Basso","sequence":"additional","affiliation":[{"name":"Center for Biostructural Imaging of Neurodegeneration, Department of Experimental Neurodegeneration, University Medical Center G\u00f6ttingen, 37075 G\u00f6ttingen, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ernst-Martin","family":"F\u00fcchtbauer","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology and Genetics, Aarhus University, 8000 Aarhus, Denmark"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Annette","family":"F\u00fcchtbauer","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology and Genetics, Aarhus University, 8000 Aarhus, Denmark"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tiago Fleming","family":"Outeiro","sequence":"additional","affiliation":[{"name":"Center for Biostructural Imaging of Neurodegeneration, Department of Experimental Neurodegeneration, University Medical Center G\u00f6ttingen, 37075 G\u00f6ttingen, Germany"},{"name":"Max Planck Institute for Multidisciplinary Sciences, 37075 G\u00f6ttingen, Germany"},{"name":"Translational and Clinical Research Institute, Faculty of Medical Sciences, Newcastle University, Framlington Place, Newcastle Upon Tyne NE2 4HH, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4439-9020","authenticated-orcid":false,"given":"Poul Henning","family":"Jensen","sequence":"additional","affiliation":[{"name":"DANDRITE, Department of Biomedicine, Aarhus University, 8000 Aarhus, Denmark"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9989-0207","authenticated-orcid":false,"given":"Torben","family":"Moos","sequence":"additional","affiliation":[{"name":"Neurobiology Research and Drug Delivery, Department of Health Science and Technology, Aalborg University, 9220 Aalborg, Denmark"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,11,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Upcott, M., Chaprov, K.D., and Buchman, V.L. (2021). Toward a Disease-Modifying Therapy of Alpha-Synucleinopathies: New Molecules and New Approaches Came into the Limelight. Molecules, 26.","DOI":"10.3390\/molecules26237351"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"29739","DOI":"10.1074\/jbc.M600933200","article-title":"Phosphorylation of Ser-129 Is the Dominant Pathological Modification of Alpha-Synuclein in Familial and Sporadic Lewy Body Disease","volume":"281","author":"Anderson","year":"2006","journal-title":"J. Biol. Chem."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Elfarrash, S., Jensen, N.M., Ferreira, N., Schmidt, S.I., Gregersen, E., Vestergaard, M.V., Nabavi, S., Meyer, M., and Jensen, P.H. (2021). Polo-like Kinase 2 Inhibition Reduces Serine-129 Phosphorylation of Physiological Nuclear Alpha-Synuclein but Not of the Aggregated Alpha-Synuclein. PLoS ONE, 16.","DOI":"10.1101\/2021.05.21.445104"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"160","DOI":"10.1038\/ncb748","article-title":"Alpha-Synuclein Is Phosphorylated in Synucleinopathy Lesions","volume":"4","author":"Fujiwara","year":"2002","journal-title":"Nat. Cell Biol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1186\/s40478-017-0452-6","article-title":"Mechanisms Underlying Extensive Ser129-Phosphorylation in \u03b1-Synuclein Aggregates","volume":"5","author":"Arawaka","year":"2017","journal-title":"Acta Neuropathol. Commun."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"16895","DOI":"10.1074\/jbc.M800747200","article-title":"Phosphorylation at Ser-129 but Not the Phosphomimics S129E\/D Inhibits the Fibrillation of Alpha-Synuclein","volume":"283","author":"Paleologou","year":"2008","journal-title":"J. Biol. Chem."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1093\/hmg\/ddy326","article-title":"Nuclear Localization and Phosphorylation Modulate Pathological Effects of Alpha-Synuclein","volume":"28","author":"Pinho","year":"2019","journal-title":"Hum. Mol. Genet."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1203","DOI":"10.1021\/cn5002254","article-title":"Efficient Modification of Alpha-Synuclein Serine 129 by Protein Kinase CK1 Requires Phosphorylation of Tyrosine 125 as a Priming Event","volume":"5","author":"Kosten","year":"2014","journal-title":"ACS Chem. Neurosci."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Kleinknecht, A., Popova, B., L\u00e1zaro, D.F., Pinho, R., Valerius, O., Outeiro, T.F., and Braus, G.H. (2016). C-Terminal Tyrosine Residue Modifications Modulate the Protective Phosphorylation of Serine 129 of \u03b1-Synuclein in a Yeast Model of Parkinson\u2019s Disease. PLoS Genet., 12.","DOI":"10.1371\/journal.pgen.1006098"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2123","DOI":"10.1074\/mcp.M800116-MCP200","article-title":"Proteomics Analysis Identifies Phosphorylation-Dependent Alpha-Synuclein Protein Interactions","volume":"7","author":"McFarland","year":"2008","journal-title":"Mol. Cell. Proteom."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1262","DOI":"10.1002\/pro.3412","article-title":"Exploring the Role of Post-Translational Modifications in Regulating \u03b1-Synuclein Interactions by Studying the Effects of Phosphorylation on Nanobody Binding","volume":"27","author":"Guilliams","year":"2018","journal-title":"Protein Sci."},{"key":"ref_12","first-page":"3257","article-title":"Tyrosine and Serine Phosphorylation of Alpha-Synuclein Have Opposing Effects on Neurotoxicity and Soluble Oligomer Formation","volume":"119","author":"Chen","year":"2009","journal-title":"J. Clin. Investig."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"182","DOI":"10.1186\/s40478-021-01281-9","article-title":"Tyrosine 136 Phosphorylation of \u03b1-Synuclein Aggregates in the Lewy Body Dementia Brain: Involvement of Serine 129 Phosphorylation by Casein Kinase 2","volume":"9","author":"Sano","year":"2021","journal-title":"Acta Neuropathol. Commun."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"617","DOI":"10.3390\/biom5020617","article-title":"Posttranslational Modifications and Clearing of \u03b1-Synuclein Aggregates in Yeast","volume":"5","author":"Popova","year":"2015","journal-title":"Biomolecules"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"568","DOI":"10.1002\/jnr.10231","article-title":"Differential Neuropathological Alterations in Transgenic Mice Expressing Alpha-Synuclein from the Platelet-Derived Growth Factor and Thy-1 Promoters","volume":"68","author":"Rockenstein","year":"2002","journal-title":"J. Neurosci. Res."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1046\/j.1471-4159.2001.00364.x","article-title":"Parkin Is Associated with Cellular Vesicles","volume":"78","author":"Kubo","year":"2001","journal-title":"J. Neurochem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"21500","DOI":"10.1074\/jbc.M000099200","article-title":"Microtubule-Associated Protein 1B Is a Component of Cortical Lewy Bodies and Binds Alpha-Synuclein Filaments","volume":"275","author":"Jensen","year":"2000","journal-title":"J. Biol. Chem."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"108","DOI":"10.1016\/j.nbd.2015.03.013","article-title":"Neurodegeneration with Inflammation Is Accompanied by Accumulation of Iron and Ferritin in Microglia and Neurons","volume":"81","author":"Thomsen","year":"2015","journal-title":"Neurobiol. Dis."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"676","DOI":"10.1038\/nmeth.2019","article-title":"Fiji: An Open-Source Platform for Biological-Image Analysis","volume":"9","author":"Schindelin","year":"2012","journal-title":"Nat Methods"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Berkhoudt Lassen, L., Gregersen, E., Kathrine Isager, A., Betzer, C., Hahn Kofoed, R., and Henning Jensen, P. (2018). ELISA Method to Detect \u03b1-Synuclein Oligomers in Cell and Animal Models. PLoS ONE, 13.","DOI":"10.1371\/journal.pone.0196056"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1002\/(SICI)1097-4547(19960715)45:2<161::AID-JNR8>3.0.CO;2-8","article-title":"OLN-93: A New Permanent Oligodendroglia Cell Line Derived From Primary Rat Brain Glial Cultures","volume":"45","author":"Heinrich","year":"1996","journal-title":"J. Neurosci. Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"10211","DOI":"10.1074\/jbc.M809671200","article-title":"Alpha-Synuclein Aggregation and Ser-129 Phosphorylation-Dependent Cell Death in Oligodendroglial Cells","volume":"284","author":"Kragh","year":"2009","journal-title":"J. Biol. Chem."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1016\/j.nbd.2013.12.002","article-title":"Prodegenerative I\u03baB\u03b1 Expression in Oligodendroglial \u03b1-Synuclein Models of Multiple System Atrophy","volume":"63","author":"Kragh","year":"2014","journal-title":"Neurobiol. Dis."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1469","DOI":"10.1016\/j.cell.2016.11.018","article-title":"Gut Microbiota Regulate Motor Deficits and Neuroinflammation in a Model of Parkinson\u2019s Disease","volume":"167","author":"Sampson","year":"2016","journal-title":"Cell"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Rabl, R., Breitschaedel, C., Flunkert, S., Duller, S., Amschl, D., Neddens, J., Niederkofler, V., Rockenstein, E., Masliah, E., and Roemer, H. (2017). Early Start of Progressive Motor Deficits in Line 61 \u03b1-Synuclein Transgenic Mice. BMC Neurosci., 18.","DOI":"10.1186\/s12868-017-0341-8"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1186\/1743-0003-2-20","article-title":"Gait Dynamics in Mouse Models of Parkinson\u2019s Disease and Huntington\u2019s Disease","volume":"2","author":"Amende","year":"2005","journal-title":"J. Neuroeng. Rehabil."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1016\/S0165-0270(01)00485-X","article-title":"A Simple Method to Measure Stride Length as an Index of Nigrostriatal Dysfunction in Mice","volume":"113","author":"Fernagut","year":"2002","journal-title":"J. Neurosci. Methods"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1440","DOI":"10.1126\/science.aan6160","article-title":"Structural Basis of Membrane Disruption and Cellular Toxicity by \u03b1-Synuclein Oligomers","volume":"358","author":"Fusco","year":"2017","journal-title":"Science"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"23179","DOI":"10.1074\/jbc.M802223200","article-title":"Serine 129 Phosphorylation of Alpha-Synuclein Induces Unfolded Protein Response-Mediated Cell Death","volume":"283","author":"Sugeno","year":"2008","journal-title":"J. Biol. Chem."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"425","DOI":"10.1016\/S0306-4522(99)00455-8","article-title":"The Absence of Reactive Astrocytosis Is Indicative of a Unique Inflammatory Process in Parkinson\u2019s Disease","volume":"95","author":"Mirza","year":"1999","journal-title":"Neuroscience"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1777","DOI":"10.1002\/jnr.22331","article-title":"Mice Expressing the A53T Mutant Form of Human Alpha-Synuclein Exhibit Hyperactivity and Reduced Anxiety-like Behavior","volume":"88","author":"Graham","year":"2010","journal-title":"J. Neurosci. Res."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1038\/nsmb.1395","article-title":"In Situ Observation of Protein Phosphorylation by High-Resolution NMR Spectroscopy","volume":"15","author":"Selenko","year":"2008","journal-title":"Nat. Struct. Mol. Biol."}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/11\/22\/3673\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:21:27Z","timestamp":1760145687000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/11\/22\/3673"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,11,18]]},"references-count":32,"journal-issue":{"issue":"22","published-online":{"date-parts":[[2022,11]]}},"alternative-id":["cells11223673"],"URL":"https:\/\/doi.org\/10.3390\/cells11223673","relation":{},"ISSN":["2073-4409"],"issn-type":[{"type":"electronic","value":"2073-4409"}],"subject":[],"published":{"date-parts":[[2022,11,18]]}}}