Modulation of Agrin and RhoA Pathways Ameliorates Movement Defects and Synapse Morphology in MYO9A-Depleted Zebrafish
Congenital myasthenic syndromes (CMS) are a group of rare, inherited disorders characterised by impaired function of the neuromuscular junction (NMJ). This is due to defects in one of the many proteins associated with the NMJ. In three patients with CMS, missense mutations in a gene encoding an unco...
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MDPI AG
2019-08-01
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| Online Access: | https://www.mdpi.com/2073-4409/8/8/848 |
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| author | Emily O’Connor George Cairns Sally Spendiff David Burns Stefan Hettwer Armin Mäder Juliane Müller Rita Horvath Clarke Slater Andreas Roos Hanns Lochmüller |
| author_facet | Emily O’Connor George Cairns Sally Spendiff David Burns Stefan Hettwer Armin Mäder Juliane Müller Rita Horvath Clarke Slater Andreas Roos Hanns Lochmüller |
| author_sort | Emily O’Connor |
| collection | DOAJ |
| description | Congenital myasthenic syndromes (CMS) are a group of rare, inherited disorders characterised by impaired function of the neuromuscular junction (NMJ). This is due to defects in one of the many proteins associated with the NMJ. In three patients with CMS, missense mutations in a gene encoding an unconventional myosin protein, MYO9A, were identified as likely causing their disorder. Preliminary studies revealed a potential involvement of the RhoA/ROCK pathway and of a key NMJ protein, agrin, in the pathophysiology of MYO9A-depletion. In this study, a CRISPR/Cas9 approach was used to generate genetic mutants of <i>MYO9A</i> zebrafish orthologues, <i>myo9aa/ab,</i> to expand and refine the morphological analysis of the NMJ. Injection of NT1654, a synthetic agrin fragment compound, improved NMJ structure and zebrafish movement in the absence of Myo9aa/ab. In addition, treatment of zebrafish with fasudil, a ROCK inhibitor, also provided improvements to the morphology of NMJs in early development, as well as rescuing movement defects, but not to the same extent as NT1654 and not at later time points. Therefore, this study highlights a role for MYO9A at the NMJ, the first unconventional myosin motor protein associated with a neuromuscular disease, and provides a potential mechanism of action of MYO9A-pathophysiology. |
| first_indexed | 2024-03-12T07:34:05Z |
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| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-12T07:34:05Z |
| publishDate | 2019-08-01 |
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| spelling | doaj.art-58e2aa45b59e444297f9c29f803bb62e2023-09-02T21:34:40ZengMDPI AGCells2073-44092019-08-018884810.3390/cells8080848cells8080848Modulation of Agrin and RhoA Pathways Ameliorates Movement Defects and Synapse Morphology in MYO9A-Depleted ZebrafishEmily O’Connor0George Cairns1Sally Spendiff2David Burns3Stefan Hettwer4Armin Mäder5Juliane Müller6Rita Horvath7Clarke Slater8Andreas Roos9Hanns Lochmüller10John Walton Muscular Dystrophy Research Centre, MRC Centre for Neuromuscular Diseases, Institute of Genetic Medicine, Newcastle University, Newcastle upon Tyne NE1 3BZ, UKInterdisciplinary School of Health Sciences, Faculty of Health Sciences, University of Ottawa, Ottawa, ON K1H 8L1, CanadaChildren’s Hospital of Eastern Ontario Research Institute, University of Ottawa, Ottawa, ON K1H 8L1, CanadaInstitute of Cell and Molecular Biosciences, Newcastle University, Newcastle Upon Tyne NE2 4HH, UKNeurotune AG. Wagistrasse 27a, 8952 Schlieren, SwitzerlandNeurotune AG. Wagistrasse 27a, 8952 Schlieren, SwitzerlandDepartment of Clinical Neurosciences, University of Cambridge, Cambridge CB2 0QQ, UKDepartment of Clinical Neurosciences, University of Cambridge, Cambridge CB2 0QQ, UKInstitute of Neuroscience, Newcastle University, Newcastle Upon Tyne NE2 4HH, UKDepartment of Neuropediatrics, Centre for Neuromuscular Disorders in Children, University Hospital Essen, University of Duisburg-Essen, 45122 Essen, GermanyChildren’s Hospital of Eastern Ontario Research Institute, University of Ottawa, Ottawa, ON K1H 8L1, CanadaCongenital myasthenic syndromes (CMS) are a group of rare, inherited disorders characterised by impaired function of the neuromuscular junction (NMJ). This is due to defects in one of the many proteins associated with the NMJ. In three patients with CMS, missense mutations in a gene encoding an unconventional myosin protein, MYO9A, were identified as likely causing their disorder. Preliminary studies revealed a potential involvement of the RhoA/ROCK pathway and of a key NMJ protein, agrin, in the pathophysiology of MYO9A-depletion. In this study, a CRISPR/Cas9 approach was used to generate genetic mutants of <i>MYO9A</i> zebrafish orthologues, <i>myo9aa/ab,</i> to expand and refine the morphological analysis of the NMJ. Injection of NT1654, a synthetic agrin fragment compound, improved NMJ structure and zebrafish movement in the absence of Myo9aa/ab. In addition, treatment of zebrafish with fasudil, a ROCK inhibitor, also provided improvements to the morphology of NMJs in early development, as well as rescuing movement defects, but not to the same extent as NT1654 and not at later time points. Therefore, this study highlights a role for MYO9A at the NMJ, the first unconventional myosin motor protein associated with a neuromuscular disease, and provides a potential mechanism of action of MYO9A-pathophysiology.https://www.mdpi.com/2073-4409/8/8/848myosin IXaunconventional myosinNT1654fasudilneuromuscular junctionMyo9aaMyo9ab |
| spellingShingle | Emily O’Connor George Cairns Sally Spendiff David Burns Stefan Hettwer Armin Mäder Juliane Müller Rita Horvath Clarke Slater Andreas Roos Hanns Lochmüller Modulation of Agrin and RhoA Pathways Ameliorates Movement Defects and Synapse Morphology in MYO9A-Depleted Zebrafish Cells myosin IXa unconventional myosin NT1654 fasudil neuromuscular junction Myo9aa Myo9ab |
| title | Modulation of Agrin and RhoA Pathways Ameliorates Movement Defects and Synapse Morphology in MYO9A-Depleted Zebrafish |
| title_full | Modulation of Agrin and RhoA Pathways Ameliorates Movement Defects and Synapse Morphology in MYO9A-Depleted Zebrafish |
| title_fullStr | Modulation of Agrin and RhoA Pathways Ameliorates Movement Defects and Synapse Morphology in MYO9A-Depleted Zebrafish |
| title_full_unstemmed | Modulation of Agrin and RhoA Pathways Ameliorates Movement Defects and Synapse Morphology in MYO9A-Depleted Zebrafish |
| title_short | Modulation of Agrin and RhoA Pathways Ameliorates Movement Defects and Synapse Morphology in MYO9A-Depleted Zebrafish |
| title_sort | modulation of agrin and rhoa pathways ameliorates movement defects and synapse morphology in myo9a depleted zebrafish |
| topic | myosin IXa unconventional myosin NT1654 fasudil neuromuscular junction Myo9aa Myo9ab |
| url | https://www.mdpi.com/2073-4409/8/8/848 |
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