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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Main Authors: Borys Olifirov, Oleksandra Fedchenko, Alexandr Dovgan, Daria Babets, Volodymyr Krotov, Volodymyr Cherkas, Pavel Belan
Format: Article
Language:English
Published: Bio-protocol LLC 2025-06-01
Series:Bio-Protocol
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
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publisher Bio-protocol LLC
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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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