Electrophysiological properties of mouse and epitope-tagged human cardiac sodium channel Nav1.5 expressed in HEK293 cells [v2; ref status: indexed, http://f1000r.es/10d]
Background: The pore-forming subunit of the cardiac sodium channel, Nav1.5, has been previously found to be mutated in genetically determined arrhythmias. Nav1.5 associates with many proteins that regulate its function and cellular localisation. In order to identify more in situ Nav1.5 interacting p...
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| Format: | Article |
| Language: | English |
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F1000 Research Ltd
2013-04-01
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| Online Access: | http://f1000research.com/articles/2-48/v2 |
| _version_ | 1818134052873437184 |
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| author | Katja Reinhard Jean-Sébastien Rougier Jakob Ogrodnik Hugues Abriel |
| author_facet | Katja Reinhard Jean-Sébastien Rougier Jakob Ogrodnik Hugues Abriel |
| author_sort | Katja Reinhard |
| collection | DOAJ |
| description | Background: The pore-forming subunit of the cardiac sodium channel, Nav1.5, has been previously found to be mutated in genetically determined arrhythmias. Nav1.5 associates with many proteins that regulate its function and cellular localisation. In order to identify more in situ Nav1.5 interacting proteins, genetically-modified mice with a high-affinity epitope in the sequence of Nav1.5 can be generated. Methods: In this short study, we (1) compared the biophysical properties of the sodium current (INa) generated by the mouse Nav1.5 (mNav1.5) and human Nav1.5 (hNav1.5) constructs that were expressed in HEK293 cells, and (2) investigated the possible alterations of the biophysical properties of the human Nav1.5 construct that was modified with specific epitopes. Results: The biophysical properties of mNav1.5 were similar to the human homolog. Addition of epitopes either up-stream of the N-terminus of hNav1.5 or in the extracellular loop between the S5 and S6 transmembrane segments of domain 1, significantly decreased the amount of INa and slightly altered its biophysical properties. Adding green fluorescent protein (GFP) to the N-terminus did not modify any of the measured biophysical properties of hNav1.5. Conclusions: These findings have to be taken into account when planning to generate genetically-modified mouse models that harbour specific epitopes in the gene encoding mNav1.5. |
| first_indexed | 2024-12-11T09:02:29Z |
| format | Article |
| id | doaj.art-e5c86efb9c7546bd9d6ff46648ed5f53 |
| institution | Directory Open Access Journal |
| issn | 2046-1402 |
| language | English |
| last_indexed | 2024-12-11T09:02:29Z |
| publishDate | 2013-04-01 |
| publisher | F1000 Research Ltd |
| record_format | Article |
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| spelling | doaj.art-e5c86efb9c7546bd9d6ff46648ed5f532022-12-22T01:13:43ZengF1000 Research LtdF1000Research2046-14022013-04-01210.12688/f1000research.2-48.v21309Electrophysiological properties of mouse and epitope-tagged human cardiac sodium channel Nav1.5 expressed in HEK293 cells [v2; ref status: indexed, http://f1000r.es/10d]Katja Reinhard0Jean-Sébastien Rougier1Jakob Ogrodnik2Hugues Abriel3Current address: Centre for Integrative Neuroscience, University of Tübingen, Tübingen, 72076, GermanyDepartment of Clinical Research, University of Bern, Bern, 3010, SwitzerlandDepartment of Clinical Research, University of Bern, Bern, 3010, SwitzerlandDepartment of Clinical Research, University of Bern, Bern, 3010, SwitzerlandBackground: The pore-forming subunit of the cardiac sodium channel, Nav1.5, has been previously found to be mutated in genetically determined arrhythmias. Nav1.5 associates with many proteins that regulate its function and cellular localisation. In order to identify more in situ Nav1.5 interacting proteins, genetically-modified mice with a high-affinity epitope in the sequence of Nav1.5 can be generated. Methods: In this short study, we (1) compared the biophysical properties of the sodium current (INa) generated by the mouse Nav1.5 (mNav1.5) and human Nav1.5 (hNav1.5) constructs that were expressed in HEK293 cells, and (2) investigated the possible alterations of the biophysical properties of the human Nav1.5 construct that was modified with specific epitopes. Results: The biophysical properties of mNav1.5 were similar to the human homolog. Addition of epitopes either up-stream of the N-terminus of hNav1.5 or in the extracellular loop between the S5 and S6 transmembrane segments of domain 1, significantly decreased the amount of INa and slightly altered its biophysical properties. Adding green fluorescent protein (GFP) to the N-terminus did not modify any of the measured biophysical properties of hNav1.5. Conclusions: These findings have to be taken into account when planning to generate genetically-modified mouse models that harbour specific epitopes in the gene encoding mNav1.5.http://f1000research.com/articles/2-48/v2Animal GeneticsCardiovascular Physiology/Circulation |
| spellingShingle | Katja Reinhard Jean-Sébastien Rougier Jakob Ogrodnik Hugues Abriel Electrophysiological properties of mouse and epitope-tagged human cardiac sodium channel Nav1.5 expressed in HEK293 cells [v2; ref status: indexed, http://f1000r.es/10d] F1000Research Animal Genetics Cardiovascular Physiology/Circulation |
| title | Electrophysiological properties of mouse and epitope-tagged human cardiac sodium channel Nav1.5 expressed in HEK293 cells [v2; ref status: indexed, http://f1000r.es/10d] |
| title_full | Electrophysiological properties of mouse and epitope-tagged human cardiac sodium channel Nav1.5 expressed in HEK293 cells [v2; ref status: indexed, http://f1000r.es/10d] |
| title_fullStr | Electrophysiological properties of mouse and epitope-tagged human cardiac sodium channel Nav1.5 expressed in HEK293 cells [v2; ref status: indexed, http://f1000r.es/10d] |
| title_full_unstemmed | Electrophysiological properties of mouse and epitope-tagged human cardiac sodium channel Nav1.5 expressed in HEK293 cells [v2; ref status: indexed, http://f1000r.es/10d] |
| title_short | Electrophysiological properties of mouse and epitope-tagged human cardiac sodium channel Nav1.5 expressed in HEK293 cells [v2; ref status: indexed, http://f1000r.es/10d] |
| title_sort | electrophysiological properties of mouse and epitope tagged human cardiac sodium channel nav1 5 expressed in hek293 cells v2 ref status indexed http f1000r es 10d |
| topic | Animal Genetics Cardiovascular Physiology/Circulation |
| url | http://f1000research.com/articles/2-48/v2 |
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