Identification of Neurons Containing Calcium-Permeable AMPA and Kainate Receptors Using Ca2+ Imaging
Calcium-permeable AMPA receptors (CP-AMPARs) and kainate receptors (CP-KARs) play crucial roles in synaptic plasticity and are implicated in various neurological processes. Current methods for identifying neurons expressing these receptors, such as electrophysiological recordings and immunostaining,...
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| Format: | Article |
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Bio-protocol LLC
2025-02-01
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| Series: | Bio-Protocol |
| Online Access: | https://bio-protocol.org/en/bpdetail?id=5199&type=0 |
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| author | Sergei Gaidin Artem Kosenkov Valery Zinchenko Bakytzhan Kairat Arailim Malibayeva Sultan Tuleukhanov |
| author_facet | Sergei Gaidin Artem Kosenkov Valery Zinchenko Bakytzhan Kairat Arailim Malibayeva Sultan Tuleukhanov |
| author_sort | Sergei Gaidin |
| collection | DOAJ |
| description | Calcium-permeable AMPA receptors (CP-AMPARs) and kainate receptors (CP-KARs) play crucial roles in synaptic plasticity and are implicated in various neurological processes. Current methods for identifying neurons expressing these receptors, such as electrophysiological recordings and immunostaining, have limitations in throughput or inability to distinguish functional receptors. This protocol describes a novel approach for the vital identification of neurons containing CP-AMPARs and CP-KARs using calcium imaging. The method involves loading neurons with Fura-2 AM, a calcium-sensitive fluorescent probe, KCl application to identify all neurons, and further addition of specific AMPAR agonists (e.g., 5-fluorowillardiine) in the presence of voltage-gated calcium channel blockers and NMDAR/KAR antagonists to identify CP-AMPAR-containing neurons. CP-KAR-containing neurons are identified using domoic acid applications in the presence and absence of NASPM (a CP-AMPAR antagonist). This technique offers several advantages over existing methods, including the ability to assess large neuronal populations simultaneously, distinguish between different receptor types, and provide functional information about CP-AMPAR and CP-KAR expression in living neurons, making it a valuable tool for studying synaptic plasticity and neurological disorders. |
| format | Article |
| id | doaj-art-e35cb10647194d639405390389fa0c99 |
| institution | Kabale University |
| issn | 2331-8325 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | Bio-protocol LLC |
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| series | Bio-Protocol |
| spelling | doaj-art-e35cb10647194d639405390389fa0c992025-02-07T08:16:46ZengBio-protocol LLCBio-Protocol2331-83252025-02-0115310.21769/BioProtoc.5199Identification of Neurons Containing Calcium-Permeable AMPA and Kainate Receptors Using Ca2+ ImagingSergei Gaidin0Artem Kosenkov1Valery Zinchenko2Bakytzhan Kairat3Arailim Malibayeva4Sultan Tuleukhanov5Federal Research Center “Pushchino Scientific Center for Biological Research of the Russian Academy of Sciences”, Institute of Cell Biophysics of the Russian Academy of Sciences, Pushchino, RussiaFederal Research Center “Pushchino Scientific Center for Biological Research of the Russian Academy of Sciences”, Institute of Cell Biophysics of the Russian Academy of Sciences, Pushchino, RussiaFederal Research Center “Pushchino Scientific Center for Biological Research of the Russian Academy of Sciences”, Institute of Cell Biophysics of the Russian Academy of Sciences, Pushchino, RussiaLaboratory of Biophysics, Chronobiology and Biomedicine, Faculty of Biology and Biotechnology, Al-Farabi Kazakh National University, Almaty, KazakhstanLaboratory of Biophysics, Chronobiology and Biomedicine, Faculty of Biology and Biotechnology, Al-Farabi Kazakh National University, Almaty, KazakhstanLaboratory of Biophysics, Chronobiology and Biomedicine, Faculty of Biology and Biotechnology, Al-Farabi Kazakh National University, Almaty, KazakhstanCalcium-permeable AMPA receptors (CP-AMPARs) and kainate receptors (CP-KARs) play crucial roles in synaptic plasticity and are implicated in various neurological processes. Current methods for identifying neurons expressing these receptors, such as electrophysiological recordings and immunostaining, have limitations in throughput or inability to distinguish functional receptors. This protocol describes a novel approach for the vital identification of neurons containing CP-AMPARs and CP-KARs using calcium imaging. The method involves loading neurons with Fura-2 AM, a calcium-sensitive fluorescent probe, KCl application to identify all neurons, and further addition of specific AMPAR agonists (e.g., 5-fluorowillardiine) in the presence of voltage-gated calcium channel blockers and NMDAR/KAR antagonists to identify CP-AMPAR-containing neurons. CP-KAR-containing neurons are identified using domoic acid applications in the presence and absence of NASPM (a CP-AMPAR antagonist). This technique offers several advantages over existing methods, including the ability to assess large neuronal populations simultaneously, distinguish between different receptor types, and provide functional information about CP-AMPAR and CP-KAR expression in living neurons, making it a valuable tool for studying synaptic plasticity and neurological disorders.https://bio-protocol.org/en/bpdetail?id=5199&type=0 |
| spellingShingle | Sergei Gaidin Artem Kosenkov Valery Zinchenko Bakytzhan Kairat Arailim Malibayeva Sultan Tuleukhanov Identification of Neurons Containing Calcium-Permeable AMPA and Kainate Receptors Using Ca2+ Imaging Bio-Protocol |
| title | Identification of Neurons Containing Calcium-Permeable AMPA and Kainate Receptors Using Ca2+ Imaging |
| title_full | Identification of Neurons Containing Calcium-Permeable AMPA and Kainate Receptors Using Ca2+ Imaging |
| title_fullStr | Identification of Neurons Containing Calcium-Permeable AMPA and Kainate Receptors Using Ca2+ Imaging |
| title_full_unstemmed | Identification of Neurons Containing Calcium-Permeable AMPA and Kainate Receptors Using Ca2+ Imaging |
| title_short | Identification of Neurons Containing Calcium-Permeable AMPA and Kainate Receptors Using Ca2+ Imaging |
| title_sort | identification of neurons containing calcium permeable ampa and kainate receptors using ca2 imaging |
| url | https://bio-protocol.org/en/bpdetail?id=5199&type=0 |
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