Local Iontophoretic Application for Pharmacological Induction of Long-Term Synaptic Depression
Long-term depression (LTD), a key form of synaptic plasticity, is typically induced through regulated Ca2+ entry via NMDA receptors and achieved by prolonged (up to hundreds of seconds) low-frequency presynaptic stimulation or bath application of NMDA receptor agonists. Electrophysiological approach...
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Bio-protocol LLC
2025-06-01
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Online Access: | https://bio-protocol.org/en/bpdetail?id=5338&type=0 |
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author | Borys Olifirov Oleksandra Fedchenko Alexandr Dovgan Daria Babets Volodymyr Krotov Volodymyr Cherkas Pavel Belan |
author_facet | Borys Olifirov Oleksandra Fedchenko Alexandr Dovgan Daria Babets Volodymyr Krotov Volodymyr Cherkas Pavel Belan |
author_sort | Borys Olifirov |
collection | DOAJ |
description | Long-term depression (LTD), a key form of synaptic plasticity, is typically induced through regulated Ca2+ entry via NMDA receptors and achieved by prolonged (up to hundreds of seconds) low-frequency presynaptic stimulation or bath application of NMDA receptor agonists. Electrophysiological approach to LTD induction requires specialized equipment, while bath applications limit productivity, as only one neuron per sample may be recorded. Here, we present a simple and effective protocol for pharmacological modeling of LTD in primary cultured neurons. This approach relies on highly localized iontophoretic application of NMDA, which induces LTD in individual cells, enhancing experimental throughput. We have analyzed spatio-temporal patterns of iontophoretic drug delivery and demonstrated how this technique may be combined with electrophysiological and live-cell imaging approaches to investigate LTD-related changes in synaptic strength and Ca2+-dependent signaling of neuronal Ca2+ sensor proteins. |
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institution | Matheson Library |
issn | 2331-8325 |
language | English |
publishDate | 2025-06-01 |
publisher | Bio-protocol LLC |
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series | Bio-Protocol |
spelling | doaj-art-401027f1b60f4cbf84a0fdb47e003ad72025-06-25T06:53:16ZengBio-protocol LLCBio-Protocol2331-83252025-06-01151110.21769/BioProtoc.5338Local Iontophoretic Application for Pharmacological Induction of Long-Term Synaptic DepressionBorys Olifirov0Oleksandra Fedchenko1Alexandr Dovgan2Daria Babets3Volodymyr Krotov4Volodymyr Cherkas5Pavel Belan6Dept. of Mol. Biophysics, Bogomoletz Inst. of Physiol. of NAS of Ukraine, Kyiv, UkraineLab. of Mol. Assays and Imaging, Inst. of Bioorganic Chem. PAS, Poznan, PolandDept. of Mol. Biophysics, Bogomoletz Inst. of Physiol. of NAS of Ukraine, Kyiv, UkraineLab. of Mol. Assays and Imaging, Inst. of Bioorganic Chem. PAS, Poznan, PolandDept. of Mol. Biophysics, Bogomoletz Inst. of Physiol. of NAS of Ukraine, Kyiv, UkraineChair of Biomedicine and Neurosci., Kyiv Academic Univ., Kyiv, UkraineDept. of Mol. Biophysics, Bogomoletz Inst. of Physiol. of NAS of Ukraine, Kyiv, UkraineDept. of Mol. Biophysics, Bogomoletz Inst. of Physiol. of NAS of Ukraine, Kyiv, UkraineLab. of Mol. Assays and Imaging, Inst. of Bioorganic Chem. PAS, Poznan, PolandDept. of Mol. Biophysics, Bogomoletz Inst. of Physiol. of NAS of Ukraine, Kyiv, UkraineChair of Biomedicine and Neurosci., Kyiv Academic Univ., Kyiv, UkraineLong-term depression (LTD), a key form of synaptic plasticity, is typically induced through regulated Ca2+ entry via NMDA receptors and achieved by prolonged (up to hundreds of seconds) low-frequency presynaptic stimulation or bath application of NMDA receptor agonists. Electrophysiological approach to LTD induction requires specialized equipment, while bath applications limit productivity, as only one neuron per sample may be recorded. Here, we present a simple and effective protocol for pharmacological modeling of LTD in primary cultured neurons. This approach relies on highly localized iontophoretic application of NMDA, which induces LTD in individual cells, enhancing experimental throughput. We have analyzed spatio-temporal patterns of iontophoretic drug delivery and demonstrated how this technique may be combined with electrophysiological and live-cell imaging approaches to investigate LTD-related changes in synaptic strength and Ca2+-dependent signaling of neuronal Ca2+ sensor proteins.https://bio-protocol.org/en/bpdetail?id=5338&type=0 |
spellingShingle | Borys Olifirov Oleksandra Fedchenko Alexandr Dovgan Daria Babets Volodymyr Krotov Volodymyr Cherkas Pavel Belan Local Iontophoretic Application for Pharmacological Induction of Long-Term Synaptic Depression Bio-Protocol |
title | Local Iontophoretic Application for Pharmacological Induction of Long-Term Synaptic Depression |
title_full | Local Iontophoretic Application for Pharmacological Induction of Long-Term Synaptic Depression |
title_fullStr | Local Iontophoretic Application for Pharmacological Induction of Long-Term Synaptic Depression |
title_full_unstemmed | Local Iontophoretic Application for Pharmacological Induction of Long-Term Synaptic Depression |
title_short | Local Iontophoretic Application for Pharmacological Induction of Long-Term Synaptic Depression |
title_sort | local iontophoretic application for pharmacological induction of long term synaptic depression |
url | https://bio-protocol.org/en/bpdetail?id=5338&type=0 |
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