Rapid purification and metabolomic profiling of synaptic vesicles from mammalian brain
Neurons communicate by the activity-dependent release of small-molecule neurotransmitters packaged into synaptic vesicles (SVs). Although many molecules have been identified as neurotransmitters, technical limitations have precluded a full metabolomic analysis of SV content. Here, we present a workf...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2020-10-01
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| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/59699 |
| _version_ | 1811200401534353408 |
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| author | Lynne Chantranupong Jessica L Saulnier Wengang Wang Drew R Jones Michael E Pacold Bernardo L Sabatini |
| author_facet | Lynne Chantranupong Jessica L Saulnier Wengang Wang Drew R Jones Michael E Pacold Bernardo L Sabatini |
| author_sort | Lynne Chantranupong |
| collection | DOAJ |
| description | Neurons communicate by the activity-dependent release of small-molecule neurotransmitters packaged into synaptic vesicles (SVs). Although many molecules have been identified as neurotransmitters, technical limitations have precluded a full metabolomic analysis of SV content. Here, we present a workflow to rapidly isolate SVs and to interrogate their metabolic contents at high-resolution using mass spectrometry. We validated the enrichment of glutamate in SVs of primary cortical neurons using targeted polar metabolomics. Unbiased and extensive global profiling of SVs isolated from these neurons revealed that the only detectable polar metabolites they contain are the established neurotransmitters glutamate and GABA. In addition, we adapted the approach to enable quick capture of SVs directly from brain tissue and determined the neurotransmitter profiles of diverse brain regions in a cell-type-specific manner. The speed, robustness, and precision of this method to interrogate SV contents will facilitate novel insights into the chemical basis of neurotransmission. |
| first_indexed | 2024-04-12T02:04:02Z |
| format | Article |
| id | doaj.art-d6079662b5df464a9044491aa0a94a6d |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T02:04:02Z |
| publishDate | 2020-10-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-d6079662b5df464a9044491aa0a94a6d2022-12-22T03:52:36ZengeLife Sciences Publications LtdeLife2050-084X2020-10-01910.7554/eLife.59699Rapid purification and metabolomic profiling of synaptic vesicles from mammalian brainLynne Chantranupong0https://orcid.org/0000-0001-9814-5264Jessica L Saulnier1Wengang Wang2Drew R Jones3Michael E Pacold4https://orcid.org/0000-0003-3688-2378Bernardo L Sabatini5https://orcid.org/0000-0003-0095-9177Department of Neurobiology, Howard Hughes Medical Institute, Harvard Medical School, Boston, United StatesDepartment of Neurobiology, Howard Hughes Medical Institute, Harvard Medical School, Boston, United StatesDepartment of Neurobiology, Howard Hughes Medical Institute, Harvard Medical School, Boston, United StatesNew York University School of Medicine, Metabolomics Core Resource Laboratory at NYU Langone Health, New York, United StatesDepartment of Radiation Oncology, New York University Langone Medical Center, New York, United StatesDepartment of Neurobiology, Howard Hughes Medical Institute, Harvard Medical School, Boston, United StatesNeurons communicate by the activity-dependent release of small-molecule neurotransmitters packaged into synaptic vesicles (SVs). Although many molecules have been identified as neurotransmitters, technical limitations have precluded a full metabolomic analysis of SV content. Here, we present a workflow to rapidly isolate SVs and to interrogate their metabolic contents at high-resolution using mass spectrometry. We validated the enrichment of glutamate in SVs of primary cortical neurons using targeted polar metabolomics. Unbiased and extensive global profiling of SVs isolated from these neurons revealed that the only detectable polar metabolites they contain are the established neurotransmitters glutamate and GABA. In addition, we adapted the approach to enable quick capture of SVs directly from brain tissue and determined the neurotransmitter profiles of diverse brain regions in a cell-type-specific manner. The speed, robustness, and precision of this method to interrogate SV contents will facilitate novel insights into the chemical basis of neurotransmission.https://elifesciences.org/articles/59699synaptic vesiclemetabolomicsneurotransmitters |
| spellingShingle | Lynne Chantranupong Jessica L Saulnier Wengang Wang Drew R Jones Michael E Pacold Bernardo L Sabatini Rapid purification and metabolomic profiling of synaptic vesicles from mammalian brain eLife synaptic vesicle metabolomics neurotransmitters |
| title | Rapid purification and metabolomic profiling of synaptic vesicles from mammalian brain |
| title_full | Rapid purification and metabolomic profiling of synaptic vesicles from mammalian brain |
| title_fullStr | Rapid purification and metabolomic profiling of synaptic vesicles from mammalian brain |
| title_full_unstemmed | Rapid purification and metabolomic profiling of synaptic vesicles from mammalian brain |
| title_short | Rapid purification and metabolomic profiling of synaptic vesicles from mammalian brain |
| title_sort | rapid purification and metabolomic profiling of synaptic vesicles from mammalian brain |
| topic | synaptic vesicle metabolomics neurotransmitters |
| url | https://elifesciences.org/articles/59699 |
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