CRISPR/Cas9-mediated nexilin deficiency interferes with cardiac contractile function in zebrafish in vivo
Abstract Nexilin (NEXN) plays a crucial role in stabilizing the sarcomeric Z-disk of striated muscle fibers and, when mutated, leads to dilated cardiomyopathy in humans. Due to its early neonatal lethality in mice, the detailed impact of the constitutive homozygous NEXN knockout on heart and skeleta...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2023-12-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-023-50065-9 |
| _version_ | 1797377008643080192 |
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| author | Janessa Hofeichner Bernd Martin Gahr Magdalena Huber Alena Boos Wolfgang Rottbauer Steffen Just |
| author_facet | Janessa Hofeichner Bernd Martin Gahr Magdalena Huber Alena Boos Wolfgang Rottbauer Steffen Just |
| author_sort | Janessa Hofeichner |
| collection | DOAJ |
| description | Abstract Nexilin (NEXN) plays a crucial role in stabilizing the sarcomeric Z-disk of striated muscle fibers and, when mutated, leads to dilated cardiomyopathy in humans. Due to its early neonatal lethality in mice, the detailed impact of the constitutive homozygous NEXN knockout on heart and skeletal muscle morphology and function is insufficiently investigated. Here, we characterized a constitutive homozygous CRISPR/Cas9-mediated nexn knockout zebrafish model. We found that Nexn deficient embryos developed significantly reduced cardiac contractility and under stressed conditions also impaired skeletal muscle organization whereas skeletal muscle function seemed not to be affected. Remarkably, in contrast to nexn morphants, CRISPR/Cas9 nexn −/− knockout embryos showed a milder phenotype without the development of a pronounced pericardial edema or blood congestion. nexn-specific expression analysis as well as whole transcriptome profiling suggest some degree of compensatory mechanisms. Transcripts of numerous essential sarcomeric proteins were massively induced and may mediate a sarcomere stabilizing function in nexn −/− knockout embryos. Our findings demonstrate the successful generation and characterization of a constitutive homozygous nexn knockout line enabling the detailed investigation of the role of nexn on heart and skeletal muscle development and function as well as to assess putative compensatory mechanisms induced by the loss of Nexn. |
| first_indexed | 2024-03-08T19:46:47Z |
| format | Article |
| id | doaj.art-e08620d4a73f433e931fbe5cc9844ed3 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-03-08T19:46:47Z |
| publishDate | 2023-12-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-e08620d4a73f433e931fbe5cc9844ed32023-12-24T12:18:51ZengNature PortfolioScientific Reports2045-23222023-12-0113111510.1038/s41598-023-50065-9CRISPR/Cas9-mediated nexilin deficiency interferes with cardiac contractile function in zebrafish in vivoJanessa Hofeichner0Bernd Martin Gahr1Magdalena Huber2Alena Boos3Wolfgang Rottbauer4Steffen Just5Molecular Cardiology, Department of Internal Medicine II, Ulm UniversityMolecular Cardiology, Department of Internal Medicine II, Ulm UniversityMolecular Cardiology, Department of Internal Medicine II, Ulm UniversityMolecular Cardiology, Department of Internal Medicine II, Ulm UniversityDepartment of Internal Medicine II, Ulm UniversityMolecular Cardiology, Department of Internal Medicine II, Ulm UniversityAbstract Nexilin (NEXN) plays a crucial role in stabilizing the sarcomeric Z-disk of striated muscle fibers and, when mutated, leads to dilated cardiomyopathy in humans. Due to its early neonatal lethality in mice, the detailed impact of the constitutive homozygous NEXN knockout on heart and skeletal muscle morphology and function is insufficiently investigated. Here, we characterized a constitutive homozygous CRISPR/Cas9-mediated nexn knockout zebrafish model. We found that Nexn deficient embryos developed significantly reduced cardiac contractility and under stressed conditions also impaired skeletal muscle organization whereas skeletal muscle function seemed not to be affected. Remarkably, in contrast to nexn morphants, CRISPR/Cas9 nexn −/− knockout embryos showed a milder phenotype without the development of a pronounced pericardial edema or blood congestion. nexn-specific expression analysis as well as whole transcriptome profiling suggest some degree of compensatory mechanisms. Transcripts of numerous essential sarcomeric proteins were massively induced and may mediate a sarcomere stabilizing function in nexn −/− knockout embryos. Our findings demonstrate the successful generation and characterization of a constitutive homozygous nexn knockout line enabling the detailed investigation of the role of nexn on heart and skeletal muscle development and function as well as to assess putative compensatory mechanisms induced by the loss of Nexn.https://doi.org/10.1038/s41598-023-50065-9 |
| spellingShingle | Janessa Hofeichner Bernd Martin Gahr Magdalena Huber Alena Boos Wolfgang Rottbauer Steffen Just CRISPR/Cas9-mediated nexilin deficiency interferes with cardiac contractile function in zebrafish in vivo Scientific Reports |
| title | CRISPR/Cas9-mediated nexilin deficiency interferes with cardiac contractile function in zebrafish in vivo |
| title_full | CRISPR/Cas9-mediated nexilin deficiency interferes with cardiac contractile function in zebrafish in vivo |
| title_fullStr | CRISPR/Cas9-mediated nexilin deficiency interferes with cardiac contractile function in zebrafish in vivo |
| title_full_unstemmed | CRISPR/Cas9-mediated nexilin deficiency interferes with cardiac contractile function in zebrafish in vivo |
| title_short | CRISPR/Cas9-mediated nexilin deficiency interferes with cardiac contractile function in zebrafish in vivo |
| title_sort | crispr cas9 mediated nexilin deficiency interferes with cardiac contractile function in zebrafish in vivo |
| url | https://doi.org/10.1038/s41598-023-50065-9 |
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