VEGF Axonal Transport Dependent on Kinesin-1B and Microtubules Dynamics
Axon-transport plays an important role in neuronal activity and survival. Reduced endogenous VEGF can cause neuronal damage and axon degeneration. It is unknown at this time if VEGF can be transported within the axon or whether it can be released by axonal depolarization. We transfected VEGF-eGFP pl...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2017-12-01
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| Series: | Frontiers in Molecular Neuroscience |
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| Online Access: | http://journal.frontiersin.org/article/10.3389/fnmol.2017.00424/full |
| _version_ | 1818586339853991936 |
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| author | Ping Yang Xiao Sun Zeng-Wei Kou Kun-Wei Wu Ya-Lin Huang Feng-Yan Sun Feng-Yan Sun |
| author_facet | Ping Yang Xiao Sun Zeng-Wei Kou Kun-Wei Wu Ya-Lin Huang Feng-Yan Sun Feng-Yan Sun |
| author_sort | Ping Yang |
| collection | DOAJ |
| description | Axon-transport plays an important role in neuronal activity and survival. Reduced endogenous VEGF can cause neuronal damage and axon degeneration. It is unknown at this time if VEGF can be transported within the axon or whether it can be released by axonal depolarization. We transfected VEGF-eGFP plasmids in cultured hippocampal neurons and tracked their movement in the axons by live-cell confocal imaging. Then, we co-transfected phVEGF-eGFP and kinesin-1B-DsRed vectors into neurons and combined with immunoprecipitation and two-color imaging to study the mechanism of VEGF axon-trafficking. We found that VEGF vesicles morphologically co-localized and biochemically bounded with kinesin-1B, as well as co-trafficked with it in the axons. Moreover, the capacity for axonal trafficking of VEGF was reduced by administration of nocodazole, an inhibitor of microtubules, or kinesin-1B shRNA. In addition, we found that VEGF could release from the cultured neurons under acute depolarizing stimulation with potassium chloride. Therefore, present findings suggest that neuronal VEGF is stored in the vesicles, actively released, and transported in the axons, which depends on the presence of kinesin-1B and functional microtubules. These results further help us to understand the importance of neuronal VEGF in the maintenance of neuronal activity and survival throughout life. |
| first_indexed | 2024-12-16T08:51:24Z |
| format | Article |
| id | doaj.art-71f46f48da004714a649ce56abbcdd20 |
| institution | Directory Open Access Journal |
| issn | 1662-5099 |
| language | English |
| last_indexed | 2024-12-16T08:51:24Z |
| publishDate | 2017-12-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Molecular Neuroscience |
| spelling | doaj.art-71f46f48da004714a649ce56abbcdd202022-12-21T22:37:24ZengFrontiers Media S.A.Frontiers in Molecular Neuroscience1662-50992017-12-011010.3389/fnmol.2017.00424281118VEGF Axonal Transport Dependent on Kinesin-1B and Microtubules DynamicsPing Yang0Xiao Sun1Zeng-Wei Kou2Kun-Wei Wu3Ya-Lin Huang4Feng-Yan Sun5Feng-Yan Sun6Department of Neurobiology, Institute for Biomedical Science and State Key Laboratory of Medical Neurobiology, School of Basic Medical Sciences, Shanghai Medical College of Fudan University, Shanghai, ChinaDepartment of Neurobiology, Institute for Biomedical Science and State Key Laboratory of Medical Neurobiology, School of Basic Medical Sciences, Shanghai Medical College of Fudan University, Shanghai, ChinaDepartment of Neurobiology, Institute for Biomedical Science and State Key Laboratory of Medical Neurobiology, School of Basic Medical Sciences, Shanghai Medical College of Fudan University, Shanghai, ChinaDepartment of Neurobiology, Institute for Biomedical Science and State Key Laboratory of Medical Neurobiology, School of Basic Medical Sciences, Shanghai Medical College of Fudan University, Shanghai, ChinaDepartment of Neurobiology, Institute for Biomedical Science and State Key Laboratory of Medical Neurobiology, School of Basic Medical Sciences, Shanghai Medical College of Fudan University, Shanghai, ChinaDepartment of Neurobiology, Institute for Biomedical Science and State Key Laboratory of Medical Neurobiology, School of Basic Medical Sciences, Shanghai Medical College of Fudan University, Shanghai, ChinaResearch Center on Aging and Medicine, Fudan University, Shanghai, ChinaAxon-transport plays an important role in neuronal activity and survival. Reduced endogenous VEGF can cause neuronal damage and axon degeneration. It is unknown at this time if VEGF can be transported within the axon or whether it can be released by axonal depolarization. We transfected VEGF-eGFP plasmids in cultured hippocampal neurons and tracked their movement in the axons by live-cell confocal imaging. Then, we co-transfected phVEGF-eGFP and kinesin-1B-DsRed vectors into neurons and combined with immunoprecipitation and two-color imaging to study the mechanism of VEGF axon-trafficking. We found that VEGF vesicles morphologically co-localized and biochemically bounded with kinesin-1B, as well as co-trafficked with it in the axons. Moreover, the capacity for axonal trafficking of VEGF was reduced by administration of nocodazole, an inhibitor of microtubules, or kinesin-1B shRNA. In addition, we found that VEGF could release from the cultured neurons under acute depolarizing stimulation with potassium chloride. Therefore, present findings suggest that neuronal VEGF is stored in the vesicles, actively released, and transported in the axons, which depends on the presence of kinesin-1B and functional microtubules. These results further help us to understand the importance of neuronal VEGF in the maintenance of neuronal activity and survival throughout life.http://journal.frontiersin.org/article/10.3389/fnmol.2017.00424/fullneurovascular unitneuropeptideneuromodulatoraxon transportneurotrophin |
| spellingShingle | Ping Yang Xiao Sun Zeng-Wei Kou Kun-Wei Wu Ya-Lin Huang Feng-Yan Sun Feng-Yan Sun VEGF Axonal Transport Dependent on Kinesin-1B and Microtubules Dynamics Frontiers in Molecular Neuroscience neurovascular unit neuropeptide neuromodulator axon transport neurotrophin |
| title | VEGF Axonal Transport Dependent on Kinesin-1B and Microtubules Dynamics |
| title_full | VEGF Axonal Transport Dependent on Kinesin-1B and Microtubules Dynamics |
| title_fullStr | VEGF Axonal Transport Dependent on Kinesin-1B and Microtubules Dynamics |
| title_full_unstemmed | VEGF Axonal Transport Dependent on Kinesin-1B and Microtubules Dynamics |
| title_short | VEGF Axonal Transport Dependent on Kinesin-1B and Microtubules Dynamics |
| title_sort | vegf axonal transport dependent on kinesin 1b and microtubules dynamics |
| topic | neurovascular unit neuropeptide neuromodulator axon transport neurotrophin |
| url | http://journal.frontiersin.org/article/10.3389/fnmol.2017.00424/full |
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