{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2024,7,5]],"date-time":"2024-07-05T18:42:54Z","timestamp":1720204974081},"reference-count":39,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2016,6,7]],"date-time":"2016-06-07T00:00:00Z","timestamp":1465257600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2016,6,7]],"date-time":"2016-06-07T00:00:00Z","timestamp":1465257600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>An unresolved mystery in the field of spinal muscular atrophy (SMA) is why a reduction of the ubiquitously expressed Smn protein causes defects mostly in motoneurons. We addressed the possibility that this restricted vulnerability stems from elevated Smn expression in motoneurons. To explore this, we established an <jats:italic>ex vivo<\/jats:italic> zebrafish culture system of GFP-marked motoneurons to quantitatively measure Smn protein and <jats:italic>smn<\/jats:italic> mRNA levels as well as promoter activity in motoneurons versus other cell types. Importantly, we uncovered that Smn levels are elevated in motoneurons by means of transcriptional activation. In addition, we identified the ETS family transcription factor Etv5b to be responsible for increased <jats:italic>smn<\/jats:italic> transcription in motoneurons. Moreover, we established that the additional supply of Smn protein in motoneurons is necessary for proper axonogenesis in a cell-autonomous manner. These findings demonstrate the reliance of motoneurons on more Smn, thereby adding a novel piece of evidence for their increased vulnerability under SMA conditions.<\/jats:p>","DOI":"10.1038\/srep27470","type":"journal-article","created":{"date-parts":[[2016,6,7]],"date-time":"2016-06-07T10:32:23Z","timestamp":1465295543000},"update-policy":"http:\/\/dx.doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Transcriptional enhancement of Smn levels in motoneurons is crucial for proper axon morphology in zebrafish"],"prefix":"10.1038","volume":"6","author":[{"given":"Zolt\u00e1n","family":"Spir\u00f3","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Angela","family":"Koh","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shermaine","family":"Tay","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kelvin","family":"See","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christoph","family":"Winkler","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2016,6,7]]},"reference":[{"key":"BFsrep27470_CR1","doi-asserted-by":"publisher","first-page":"276","DOI":"10.1001\/archneur.1971.00490030102014","volume":"25","author":"G Werdnig","year":"1971","unstructured":"Werdnig, G. Two early infantile hereditary cases of progressive muscular atrophy simulating dystrophy, but on a neural basis. 1891. Arch Neurol 25, 276\u2013278 (1971).","journal-title":"Arch Neurol"},{"key":"BFsrep27470_CR2","doi-asserted-by":"publisher","first-page":"460","DOI":"10.1038\/ng1207","volume":"34","author":"T Kashima","year":"2003","unstructured":"Kashima, T. & Manley, J. L. A negative element in SMN2 exon 7 inhibits splicing in spinal muscular atrophy. Nat Genet 34, 460\u2013463, doi: 10.1038\/ng1207 (2003).","journal-title":"Nat Genet"},{"key":"BFsrep27470_CR3","doi-asserted-by":"publisher","first-page":"155","DOI":"10.1016\/0092-8674(95)90460-3","volume":"80","author":"S Lefebvre","year":"1995","unstructured":"Lefebvre, S. et al. Identification and characterization of a spinal muscular atrophy-determining gene. Cell 80, 155\u2013165 (1995).","journal-title":"Cell"},{"key":"BFsrep27470_CR4","doi-asserted-by":"publisher","first-page":"1177","DOI":"10.1093\/hmg\/8.7.1177","volume":"8","author":"UR Monani","year":"1999","unstructured":"Monani, U. R. et al. A single nucleotide difference that alters splicing patterns distinguishes the SMA gene SMN1 from the copy gene SMN2. Hum Mol Genet 8, 1177\u20131183 (1999).","journal-title":"Hum Mol Genet"},{"key":"BFsrep27470_CR5","doi-asserted-by":"publisher","first-page":"5665","DOI":"10.1093\/hmg\/ddv292","volume":"24","author":"SE Gombash","year":"2015","unstructured":"Gombash, S. E. et al. SMN deficiency disrupts gastrointestinal and enteric nervous system function in mice. Hum Mol Genet 24, 5665, doi: 10.1093\/hmg\/ddv292 (2015).","journal-title":"Hum Mol Genet"},{"key":"BFsrep27470_CR6","doi-asserted-by":"publisher","first-page":"2235","DOI":"10.1093\/hmg\/ddt612","volume":"23","author":"G Hunter","year":"2014","unstructured":"Hunter, G., Aghamaleky Sarvestany, A., Roche, S. L., Symes, R. C. & Gillingwater, T. H. SMN-dependent intrinsic defects in Schwann cells in mouse models of spinal muscular atrophy. Hum Mol Genet 23, 2235\u20132250, doi: 10.1093\/hmg\/ddt612 (2014).","journal-title":"Hum Mol Genet"},{"key":"BFsrep27470_CR7","doi-asserted-by":"publisher","first-page":"5524","DOI":"10.1093\/hmg\/ddv283","volume":"24","author":"VL McGovern","year":"2015","unstructured":"McGovern, V. L. et al. SMN expression is required in motor neurons to rescue electrophysiological deficits in the SMNDelta7 mouse model of SMA. Hum Mol Genet 24, 5524\u20135541, doi: 10.1093\/hmg\/ddv283 (2015).","journal-title":"Hum Mol Genet"},{"key":"BFsrep27470_CR8","doi-asserted-by":"publisher","first-page":"8703","DOI":"10.1523\/JNEUROSCI.0204-12.2012","volume":"32","author":"TL Martinez","year":"2012","unstructured":"Martinez, T. L. et al. Survival motor neuron protein in motor neurons determines synaptic integrity in spinal muscular atrophy. J Neurosci 32, 8703\u20138715, doi: 10.1523\/JNEUROSCI.0204-12.2012 (2012).","journal-title":"J Neurosci"},{"key":"BFsrep27470_CR9","doi-asserted-by":"publisher","first-page":"e75866","DOI":"10.1371\/journal.pone.0075866","volume":"8","author":"X Paez-Colasante","year":"2013","unstructured":"Paez-Colasante, X. et al. Improvement of neuromuscular synaptic phenotypes without enhanced survival and motor function in severe spinal muscular atrophy mice selectively rescued in motor neurons. PLoS One 8, e75866, doi: 10.1371\/journal.pone.0075866 (2013).","journal-title":"PLoS One"},{"key":"BFsrep27470_CR10","doi-asserted-by":"publisher","first-page":"12005","DOI":"10.1523\/JNEUROSCI.2208-10.2010","volume":"30","author":"GH Park","year":"2010","unstructured":"Park, G. H., Maeno-Hikichi, Y., Awano, T., Landmesser, L. T. & Monani, U. R. Reduced survival of motor neuron (SMN) protein in motor neuronal progenitors functions cell autonomously to cause spinal muscular atrophy in model mice expressing the human centromeric (SMN2) gene. J Neurosci 30, 12005\u201312019, doi: 10.1523\/JNEUROSCI.2208-10.2010 (2010).","journal-title":"J Neurosci"},{"key":"BFsrep27470_CR11","doi-asserted-by":"publisher","first-page":"123","DOI":"10.1038\/nature10485","volume":"478","author":"Y Hua","year":"2011","unstructured":"Hua, Y. et al. Peripheral SMN restoration is essential for long-term rescue of a severe spinal muscular atrophy mouse model. Nature 478, 123\u2013126, doi: 10.1038\/nature10485 (2011).","journal-title":"Nature"},{"key":"BFsrep27470_CR12","doi-asserted-by":"publisher","first-page":"346","DOI":"10.1093\/hmg\/ddu447","volume":"24","author":"T Hao le","year":"2015","unstructured":"Hao le, T., Duy, P. Q., Jontes, J. D. & Beattie, C. E. Motoneuron development influences dorsal root ganglia survival and Schwann cell development in a vertebrate model of spinal muscular atrophy. Human molecular genetics 24, 346\u2013360, doi: 10.1093\/hmg\/ddu447 (2015).","journal-title":"Human molecular genetics"},{"key":"BFsrep27470_CR13","doi-asserted-by":"publisher","first-page":"919","DOI":"10.1083\/jcb.200303168","volume":"162","author":"ML McWhorter","year":"2003","unstructured":"McWhorter, M. L., Monani, U. R., Burghes, A. H. & Beattie, C. E. Knockdown of the survival motor neuron (Smn) protein in zebrafish causes defects in motor axon outgrowth and pathfinding. The Journal of cell biology 162, 919\u2013931, doi: 10.1083\/jcb.200303168 (2003).","journal-title":"The Journal of cell biology"},{"key":"BFsrep27470_CR14","doi-asserted-by":"publisher","first-page":"2320","DOI":"10.1101\/gad.342005","volume":"19","author":"C Winkler","year":"2005","unstructured":"Winkler, C. et al. Reduced U snRNP assembly causes motor axon degeneration in an animal model for spinal muscular atrophy. Genes & development 19, 2320\u20132330, doi: 10.1101\/gad.342005 (2005).","journal-title":"Genes & development"},{"key":"BFsrep27470_CR15","doi-asserted-by":"publisher","first-page":"7378","DOI":"10.1038\/ncomms8378","volume":"6","author":"J Giacomotto","year":"2015","unstructured":"Giacomotto, J., Rinkwitz, S. & Becker, T. S. Effective heritable gene knockdown in zebrafish using synthetic microRNAs. Nat Commun 6, 7378, doi: 10.1038\/ncomms8378 (2015).","journal-title":"Nat Commun"},{"key":"BFsrep27470_CR16","doi-asserted-by":"publisher","first-page":"2612","DOI":"10.1093\/hmg\/ddt110","volume":"22","author":"T Hao le","year":"2013","unstructured":"Hao le, T. et al. Temporal requirement for SMN in motoneuron development. Human molecular genetics 22, 2612\u20132625, doi: 10.1093\/hmg\/ddt110 (2013).","journal-title":"Human molecular genetics"},{"key":"BFsrep27470_CR17","doi-asserted-by":"publisher","first-page":"1754","DOI":"10.1093\/hmg\/ddt567","volume":"23","author":"K See","year":"2014","unstructured":"See, K. et al. SMN deficiency alters Nrxn2 expression and splicing in zebrafish and mouse models of spinal muscular atrophy. Hum Mol Genet 23, 1754\u20131770, doi: 10.1093\/hmg\/ddt567 (2014).","journal-title":"Hum Mol Genet"},{"key":"BFsrep27470_CR18","doi-asserted-by":"publisher","first-page":"19348","DOI":"10.1073\/pnas.1319280110","volume":"110","author":"Z Zhang","year":"2013","unstructured":"Zhang, Z. et al. Dysregulation of synaptogenesis genes antecedes motor neuron pathology in spinal muscular atrophy. Proc Natl Acad Sci USA 110, 19348\u201319353, doi: 10.1073\/pnas.1319280110 (2013).","journal-title":"Proc Natl Acad Sci USA"},{"key":"BFsrep27470_CR19","doi-asserted-by":"publisher","first-page":"1430","DOI":"10.1080\/15476286.2014.996494","volume":"11","author":"Q Huo","year":"2014","unstructured":"Huo, Q. et al. Splicing changes in SMA mouse motoneurons and SMN-depleted neuroblastoma cells: evidence for involvement of splicing regulatory proteins. RNA Biol 11, 1430\u20131446, doi: 10.1080\/15476286.2014.996494 (2014).","journal-title":"RNA Biol"},{"key":"BFsrep27470_CR20","doi-asserted-by":"publisher","first-page":"801","DOI":"10.1083\/jcb.200304128","volume":"163","author":"W Rossoll","year":"2003","unstructured":"Rossoll, W. et al. Smn, the spinal muscular atrophy-determining gene product, modulates axon growth and localization of beta-actin mRNA in growth cones of motoneurons. The Journal of cell biology 163, 801\u2013812, doi: 10.1083\/jcb.200304128 (2003).","journal-title":"The Journal of cell biology"},{"key":"BFsrep27470_CR21","doi-asserted-by":"publisher","first-page":"93","DOI":"10.1093\/hmg\/11.1.93","volume":"11","author":"W Rossoll","year":"2002","unstructured":"Rossoll, W. et al. Specific interaction of Smn, the spinal muscular atrophy determining gene product, with hnRNP-R and gry-rbp\/hnRNP-Q: a role for Smn in RNA processing in motor axons? Hum Mol Genet 11, 93\u2013105 (2002).","journal-title":"Hum Mol Genet"},{"key":"BFsrep27470_CR22","doi-asserted-by":"publisher","first-page":"e110846","DOI":"10.1371\/journal.pone.0110846","volume":"9","author":"B Dombert","year":"2014","unstructured":"Dombert, B., Sivadasan, R., Simon, C. M., Jablonka, S. & Sendtner, M. Presynaptic localization of Smn and hnRNP R in axon terminals of embryonic and postnatal mouse motoneurons. PLoS One 9, e110846, doi: 10.1371\/journal.pone.0110846 (2014).","journal-title":"PLoS One"},{"key":"BFsrep27470_CR23","doi-asserted-by":"publisher","first-page":"1789","DOI":"10.1261\/rna.047373.114","volume":"20","author":"L Saal","year":"2014","unstructured":"Saal, L., Briese, M., Kneitz, S., Glinka, M. & Sendtner, M. Subcellular transcriptome alterations in a cell culture model of spinal muscular atrophy point to widespread defects in axonal growth and presynaptic differentiation. RNA 20, 1789\u20131802, doi: 10.1261\/rna.047373.114 (2014).","journal-title":"RNA"},{"key":"BFsrep27470_CR24","doi-asserted-by":"publisher","first-page":"293","DOI":"10.1046\/j.1460-9568.1999.00428.x","volume":"11","author":"C Bechade","year":"1999","unstructured":"Bechade, C. et al. Subcellular distribution of survival motor neuron (SMN) protein: possible involvement in nucleocytoplasmic and dendritic transport. Eur J Neurosci 11, 293\u2013304 (1999).","journal-title":"Eur J Neurosci"},{"key":"BFsrep27470_CR25","doi-asserted-by":"publisher","first-page":"265","DOI":"10.1038\/ng0797-265","volume":"16","author":"S Lefebvre","year":"1997","unstructured":"Lefebvre, S. et al. Correlation between severity and SMN protein level in spinal muscular atrophy. Nat Genet 16, 265\u2013269, doi: 10.1038\/ng0797-265 (1997).","journal-title":"Nat Genet"},{"key":"BFsrep27470_CR26","doi-asserted-by":"publisher","first-page":"143","DOI":"10.1038\/nn2025","volume":"11","author":"S Kucenas","year":"2008","unstructured":"Kucenas, S. et al. CNS-derived glia ensheath peripheral nerves and mediate motor root development. Nat Neurosci 11, 143\u2013151, doi: 10.1038\/nn2025 (2008).","journal-title":"Nat Neurosci"},{"key":"BFsrep27470_CR27","doi-asserted-by":"publisher","first-page":"729","DOI":"10.1038\/sj.ejhg.5201217","volume":"12","author":"B Boda","year":"2004","unstructured":"Boda, B. et al. Survival motor neuron SMN1 and SMN2 gene promoters: identical sequences and differential expression in neurons and non-neuronal cells. Eur J Hum Genet 12, 729\u2013737, doi: 10.1038\/sj.ejhg.5201217 (2004).","journal-title":"Eur J Hum Genet"},{"key":"BFsrep27470_CR28","doi-asserted-by":"publisher","first-page":"330","DOI":"10.1016\/S0167-4781(99)00060-3","volume":"1445","author":"UR Monani","year":"1999","unstructured":"Monani, U. R., McPherson, J. D. & Burghes, A. H. Promoter analysis of the human centromeric and telomeric survival motor neuron genes (SMNC and SMNT). Biochim Biophys Acta 1445, 330\u2013336 (1999).","journal-title":"Biochim Biophys Acta"},{"key":"BFsrep27470_CR29","doi-asserted-by":"publisher","first-page":"877","DOI":"10.1016\/S0896-6273(02)00863-2","volume":"35","author":"J Livet","year":"2002","unstructured":"Livet, J. et al. ETS gene Pea3 controls the central position and terminal arborization of specific motor neuron pools. Neuron 35, 877\u2013892 (2002).","journal-title":"Neuron"},{"key":"BFsrep27470_CR30","doi-asserted-by":"publisher","first-page":"1979","DOI":"10.1101\/gr.209601","volume":"11","author":"T Kudoh","year":"2001","unstructured":"Kudoh, T. et al. A gene expression screen in zebrafish embryogenesis. Genome Res 11, 1979\u20131987, doi: 10.1101\/gr.209601 (2001).","journal-title":"Genome Res"},{"key":"BFsrep27470_CR31","doi-asserted-by":"publisher","first-page":"777","DOI":"10.1016\/j.modgep.2006.01.008","volume":"6","author":"M Roussigne","year":"2006","unstructured":"Roussigne, M. & Blader, P. Divergence in regulation of the PEA3 family of ETS transcription factors. Gene Expr Patterns 6, 777\u2013782, doi: 10.1016\/j.modgep.2006.01.008 (2006).","journal-title":"Gene Expr Patterns"},{"key":"BFsrep27470_CR32","doi-asserted-by":"publisher","first-page":"87","DOI":"10.1093\/nar\/29.1.87","volume":"29","author":"J Sprague","year":"2001","unstructured":"Sprague, J., Doerry, E., Douglas, S. & Westerfield, M. The Zebrafish Information Network (ZFIN): a resource for genetic, genomic and developmental research. Nucleic Acids Res 29, 87\u201390 (2001).","journal-title":"Nucleic Acids Res"},{"key":"BFsrep27470_CR33","doi-asserted-by":"publisher","first-page":"600","DOI":"10.1016\/j.devcel.2009.02.005","volume":"16","author":"J Mao","year":"2009","unstructured":"Mao, J., McGlinn, E., Huang, P., Tabin, C. J. & McMahon, A. P. Fgf-dependent Etv4\/5 activity is required for posterior restriction of Sonic Hedgehog and promoting outgrowth of the vertebrate limb. Dev Cell 16, 600\u2013606, doi: 10.1016\/j.devcel.2009.02.005 (2009).","journal-title":"Dev Cell"},{"key":"BFsrep27470_CR34","doi-asserted-by":"publisher","first-page":"126","DOI":"10.1128\/MCB.06077-11","volume":"32","author":"M Ruggiu","year":"2012","unstructured":"Ruggiu, M. et al. A role for SMN exon 7 splicing in the selective vulnerability of motor neurons in spinal muscular atrophy. Mol Cell Biol 32, 126\u2013138, doi: 10.1128\/MCB.06077-11 (2012).","journal-title":"Mol Cell Biol"},{"key":"BFsrep27470_CR35","doi-asserted-by":"publisher","first-page":"290","DOI":"10.1016\/j.ydbio.2012.03.001","volume":"365","author":"V Arkhipova","year":"2012","unstructured":"Arkhipova, V. et al. Characterization and regulation of the hb9\/mnx1 beta-cell progenitor specific enhancer in zebrafish. Developmental biology 365, 290\u2013302, doi: 10.1016\/j.ydbio.2012.03.001 (2012).","journal-title":"Developmental biology"},{"key":"BFsrep27470_CR36","doi-asserted-by":"publisher","first-page":"1133","DOI":"10.1038\/nprot.2006.165","volume":"1","author":"M Rembold","year":"2006","unstructured":"Rembold, M., Lahiri, K., Foulkes, N. S. & Wittbrodt, J. Transgenesis in fish: efficient selection of transgenic fish by co-injection with a fluorescent reporter construct. Nat Protoc 1, 1133\u20131139, doi: 10.1038\/nprot.2006.165 (2006).","journal-title":"Nat Protoc"},{"key":"BFsrep27470_CR37","doi-asserted-by":"publisher","first-page":"e57539","DOI":"10.1371\/journal.pone.0057539","volume":"8","author":"Z Chen","year":"2013","unstructured":"Chen, Z. et al. Primary neuron culture for nerve growth and axon guidance studies in zebrafish (Danio rerio). PLoS One 8, e57539, doi: 10.1371\/journal.pone.0057539 (2013).","journal-title":"PLoS One"},{"key":"BFsrep27470_CR38","doi-asserted-by":"publisher","first-page":"269","DOI":"10.1038\/320269a0","volume":"320","author":"JS Eisen","year":"1986","unstructured":"Eisen, J. S., Myers, P. Z. & Westerfield, M. Pathway selection by growth cones of identified motoneurones in live zebra fish embryos. Nature 320, 269\u2013271, doi: 10.1038\/320269a0 (1986).","journal-title":"Nature"},{"key":"BFsrep27470_CR39","doi-asserted-by":"publisher","first-page":"465","DOI":"10.1089\/zeb.2006.3.465","volume":"3","author":"MC Welten","year":"2006","unstructured":"Welten, M. C. et al. ZebraFISH: fluorescent in situ hybridization protocol and three-dimensional imaging of gene expression patterns. Zebrafish 3, 465\u2013476, doi: 10.1089\/zeb.2006.3.465 (2006).","journal-title":"Zebrafish"}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/srep27470.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/srep27470","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/srep27470.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,1,5]],"date-time":"2023-01-05T05:28:18Z","timestamp":1672896498000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/srep27470"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2016,6,7]]},"references-count":39,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2016,6,20]]}},"alternative-id":["BFsrep27470"],"URL":"https:\/\/doi.org\/10.1038\/srep27470","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2016,6,7]]},"assertion":[{"value":"8 December 2015","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"19 May 2016","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"7 June 2016","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing financial interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"27470"}}