Super-resolution imaging of synaptic scaffold proteins in rat hippocampal neurons
Summary: Visualizing the nano-organization of the synapse is fundamental to elucidating the structure-function relationship of the nervous system. The advent of super-resolution microscopy provides a tool to assess and quantify the dynamic organization of numerous proteins at the synapse. Here we pr...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-03-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166723000382 |
| _version_ | 1811172197422596096 |
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| author | Natalie J. Guzikowski Ege T. Kavalali |
| author_facet | Natalie J. Guzikowski Ege T. Kavalali |
| author_sort | Natalie J. Guzikowski |
| collection | DOAJ |
| description | Summary: Visualizing the nano-organization of the synapse is fundamental to elucidating the structure-function relationship of the nervous system. The advent of super-resolution microscopy provides a tool to assess and quantify the dynamic organization of numerous proteins at the synapse. Here we present a protocol assessing inhibitory synapse scaffold protein, gephyrin, in rat primary hippocampal cultures using dSTORM microscopy. We delineate the steps for artemisinin treatment, immunocytochemistry, dSTORM image acquisition, single-molecule localization, and the analysis of synaptic scaffold protein dynamics.For complete details on the use and execution of this protocol, please refer to Guzikowski and Kavalali (2022).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-04-10T17:25:47Z |
| format | Article |
| id | doaj.art-4b81fab869c14bbbad193dcaf895ebe4 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-04-10T17:25:47Z |
| publishDate | 2023-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-4b81fab869c14bbbad193dcaf895ebe42023-02-04T04:18:49ZengElsevierSTAR Protocols2666-16672023-03-0141102080Super-resolution imaging of synaptic scaffold proteins in rat hippocampal neuronsNatalie J. Guzikowski0Ege T. Kavalali1Department of Pharmacology and Vanderbilt Brain Institute, Vanderbilt University, Nashville, TN, USA; Corresponding authorDepartment of Pharmacology and Vanderbilt Brain Institute, Vanderbilt University, Nashville, TN, USA; Corresponding authorSummary: Visualizing the nano-organization of the synapse is fundamental to elucidating the structure-function relationship of the nervous system. The advent of super-resolution microscopy provides a tool to assess and quantify the dynamic organization of numerous proteins at the synapse. Here we present a protocol assessing inhibitory synapse scaffold protein, gephyrin, in rat primary hippocampal cultures using dSTORM microscopy. We delineate the steps for artemisinin treatment, immunocytochemistry, dSTORM image acquisition, single-molecule localization, and the analysis of synaptic scaffold protein dynamics.For complete details on the use and execution of this protocol, please refer to Guzikowski and Kavalali (2022).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166723000382MicroscopyNeuroscience |
| spellingShingle | Natalie J. Guzikowski Ege T. Kavalali Super-resolution imaging of synaptic scaffold proteins in rat hippocampal neurons STAR Protocols Microscopy Neuroscience |
| title | Super-resolution imaging of synaptic scaffold proteins in rat hippocampal neurons |
| title_full | Super-resolution imaging of synaptic scaffold proteins in rat hippocampal neurons |
| title_fullStr | Super-resolution imaging of synaptic scaffold proteins in rat hippocampal neurons |
| title_full_unstemmed | Super-resolution imaging of synaptic scaffold proteins in rat hippocampal neurons |
| title_short | Super-resolution imaging of synaptic scaffold proteins in rat hippocampal neurons |
| title_sort | super resolution imaging of synaptic scaffold proteins in rat hippocampal neurons |
| topic | Microscopy Neuroscience |
| url | http://www.sciencedirect.com/science/article/pii/S2666166723000382 |
| work_keys_str_mv | AT nataliejguzikowski superresolutionimagingofsynapticscaffoldproteinsinrathippocampalneurons AT egetkavalali superresolutionimagingofsynapticscaffoldproteinsinrathippocampalneurons |