Altered striatal dopamine levels in Parkinson’s disease VPS35 D620N mutant transgenic aged mice
Abstract Vacuolar protein sorting 35 (VPS35) is a major component of the retromer complex that mediates the retrograde transport of cargo proteins from endosomes to the trans-Golgi network. Mutations such as D620N in the VPS35 gene have been identified in patients with autosomal dominant Parkinson’s...
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| Format: | Article |
| Language: | English |
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BMC
2020-12-01
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| Series: | Molecular Brain |
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| Online Access: | https://doi.org/10.1186/s13041-020-00704-3 |
| _version_ | 1818620361682452480 |
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| author | Sarivin Vanan Xiaoxia Zeng Sook Yoong Chia Katarina Varnäs Mei Jiang Ke Zhang Wuan Ting Saw Parasuraman Padmanabhan Wei-Ping Yu Zhi-Dong Zhou Christer Halldin Balázs Gulyás Eng-King Tan Li Zeng |
| author_facet | Sarivin Vanan Xiaoxia Zeng Sook Yoong Chia Katarina Varnäs Mei Jiang Ke Zhang Wuan Ting Saw Parasuraman Padmanabhan Wei-Ping Yu Zhi-Dong Zhou Christer Halldin Balázs Gulyás Eng-King Tan Li Zeng |
| author_sort | Sarivin Vanan |
| collection | DOAJ |
| description | Abstract Vacuolar protein sorting 35 (VPS35) is a major component of the retromer complex that mediates the retrograde transport of cargo proteins from endosomes to the trans-Golgi network. Mutations such as D620N in the VPS35 gene have been identified in patients with autosomal dominant Parkinson’s disease (PD). However, it remains poorly understood whether and how VPS35 deficiency or mutation contributes to PD pathogenesis; specifically, the studies that have examined VPS35 thus far have differed in results and methodologies. We generated a VPS35 D620N mouse model using a Rosa26-based transgene expression platform to allow expression in a spatial manner, so as to better address these discrepancies. Here, aged (20-months-old) mice were first subjected to behavioral tests. Subsequently, DAB staining analysis of substantia nigra (SN) dopaminergic neurons with the marker for tyrosine hydroxylase (TH) was performed. Next, HPLC was used to determine dopamine levels, along with levels of its two metabolites, 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), in the striatum. Western blotting was also performed to study the levels of key proteins associated with PD. Lastly, autoradiography (ARG) evaluation of [3H]FE-PE2I binding to the striatal dopamine transporter DAT was carried out. We found that VPS35 D620N Tg mice displayed a significantly higher dopamine level than NTg counterparts. All results were then compared with that of current VPS35 studies to shed light on the disease pathogenesis. Our model allows future studies to explicitly control spatial expression of the transgene which would generate a more reliable PD phenotype. |
| first_indexed | 2024-12-16T17:52:10Z |
| format | Article |
| id | doaj.art-85c535b2d2c24c6982aab9b8d5a79aef |
| institution | Directory Open Access Journal |
| issn | 1756-6606 |
| language | English |
| last_indexed | 2024-12-16T17:52:10Z |
| publishDate | 2020-12-01 |
| publisher | BMC |
| record_format | Article |
| series | Molecular Brain |
| spelling | doaj.art-85c535b2d2c24c6982aab9b8d5a79aef2022-12-21T22:22:16ZengBMCMolecular Brain1756-66062020-12-0113111510.1186/s13041-020-00704-3Altered striatal dopamine levels in Parkinson’s disease VPS35 D620N mutant transgenic aged miceSarivin Vanan0Xiaoxia Zeng1Sook Yoong Chia2Katarina Varnäs3Mei Jiang4Ke Zhang5Wuan Ting Saw6Parasuraman Padmanabhan7Wei-Ping Yu8Zhi-Dong Zhou9Christer Halldin10Balázs Gulyás11Eng-King Tan12Li Zeng13Neural Stem Cell Research Lab, Research Department, National Neuroscience InstituteNeural Stem Cell Research Lab, Research Department, National Neuroscience InstituteNeural Stem Cell Research Lab, Research Department, National Neuroscience InstituteDepartment of Clinical Neuroscience, Psychiatry Section, Karolinska Institutet PET Centre, Karolinska Institutet, Karolinska University Hospital SolnaNeural Stem Cell Research Lab, Research Department, National Neuroscience InstituteNeural Stem Cell Research Lab, Research Department, National Neuroscience InstituteDepartment of Research, National Neuroscience InstituteLee Kong Chian School of Medicine, Nanyang Technological UniversityAnimal Gene Editing Laboratory, Biological Resource Centre, A*STARDepartment of Research, National Neuroscience InstituteDepartment of Clinical Neuroscience, Psychiatry Section, Karolinska Institutet PET Centre, Karolinska Institutet, Karolinska University Hospital SolnaDepartment of Clinical Neuroscience, Psychiatry Section, Karolinska Institutet PET Centre, Karolinska Institutet, Karolinska University Hospital SolnaDepartment of Research, National Neuroscience InstituteNeural Stem Cell Research Lab, Research Department, National Neuroscience InstituteAbstract Vacuolar protein sorting 35 (VPS35) is a major component of the retromer complex that mediates the retrograde transport of cargo proteins from endosomes to the trans-Golgi network. Mutations such as D620N in the VPS35 gene have been identified in patients with autosomal dominant Parkinson’s disease (PD). However, it remains poorly understood whether and how VPS35 deficiency or mutation contributes to PD pathogenesis; specifically, the studies that have examined VPS35 thus far have differed in results and methodologies. We generated a VPS35 D620N mouse model using a Rosa26-based transgene expression platform to allow expression in a spatial manner, so as to better address these discrepancies. Here, aged (20-months-old) mice were first subjected to behavioral tests. Subsequently, DAB staining analysis of substantia nigra (SN) dopaminergic neurons with the marker for tyrosine hydroxylase (TH) was performed. Next, HPLC was used to determine dopamine levels, along with levels of its two metabolites, 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), in the striatum. Western blotting was also performed to study the levels of key proteins associated with PD. Lastly, autoradiography (ARG) evaluation of [3H]FE-PE2I binding to the striatal dopamine transporter DAT was carried out. We found that VPS35 D620N Tg mice displayed a significantly higher dopamine level than NTg counterparts. All results were then compared with that of current VPS35 studies to shed light on the disease pathogenesis. Our model allows future studies to explicitly control spatial expression of the transgene which would generate a more reliable PD phenotype.https://doi.org/10.1186/s13041-020-00704-3Parkinson’s diseaseVPS35 D620NStriatal dopamineTransgenic miceBehavioral assay |
| spellingShingle | Sarivin Vanan Xiaoxia Zeng Sook Yoong Chia Katarina Varnäs Mei Jiang Ke Zhang Wuan Ting Saw Parasuraman Padmanabhan Wei-Ping Yu Zhi-Dong Zhou Christer Halldin Balázs Gulyás Eng-King Tan Li Zeng Altered striatal dopamine levels in Parkinson’s disease VPS35 D620N mutant transgenic aged mice Molecular Brain Parkinson’s disease VPS35 D620N Striatal dopamine Transgenic mice Behavioral assay |
| title | Altered striatal dopamine levels in Parkinson’s disease VPS35 D620N mutant transgenic aged mice |
| title_full | Altered striatal dopamine levels in Parkinson’s disease VPS35 D620N mutant transgenic aged mice |
| title_fullStr | Altered striatal dopamine levels in Parkinson’s disease VPS35 D620N mutant transgenic aged mice |
| title_full_unstemmed | Altered striatal dopamine levels in Parkinson’s disease VPS35 D620N mutant transgenic aged mice |
| title_short | Altered striatal dopamine levels in Parkinson’s disease VPS35 D620N mutant transgenic aged mice |
| title_sort | altered striatal dopamine levels in parkinson s disease vps35 d620n mutant transgenic aged mice |
| topic | Parkinson’s disease VPS35 D620N Striatal dopamine Transgenic mice Behavioral assay |
| url | https://doi.org/10.1186/s13041-020-00704-3 |
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