Cargando…
An optogenetic method for investigating presynaptic molecular regulation
While efficient methods are well established for studying postsynaptic protein regulation of glutamatergic synapses in the mammalian central nervous system, similarly efficient methods are lacking for studying proteins regulating presynaptic function. In the present study, we introduce an optical/el...
Autores principales: | , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8166971/ https://www.ncbi.nlm.nih.gov/pubmed/34059719 http://dx.doi.org/10.1038/s41598-021-90244-0 |
_version_ | 1783701604025761792 |
---|---|
author | Kay, Yuni Herring, Bruce E. |
author_facet | Kay, Yuni Herring, Bruce E. |
author_sort | Kay, Yuni |
collection | PubMed |
description | While efficient methods are well established for studying postsynaptic protein regulation of glutamatergic synapses in the mammalian central nervous system, similarly efficient methods are lacking for studying proteins regulating presynaptic function. In the present study, we introduce an optical/electrophysiological method for investigating presynaptic molecular regulation. Here, using an optogenetic approach, we selectively stimulate genetically modified presynaptic CA3 pyramidal neurons in the hippocampus and measure optically-induced excitatory postsynaptic currents produced in unmodified postsynaptic CA1 pyramidal neurons. While such use of optogenetics is not novel, previous implementation methods do not allow basic quantification of the changes in synaptic strength produced by genetic manipulations. We find that incorporating simultaneous recordings of fiber volley amplitude provides a control for optical stimulation intensity and, as a result, creates a metric of synaptic efficacy that can be compared across experimental conditions. In the present study, we utilize our new method to demonstrate that inhibition of synaptotagmin 1 expression in CA3 pyramidal neurons leads to a significant reduction in Schaffer collateral synapse function, an effect that is masked with conventional electrical stimulation. Our hope is that this method will expedite our understanding of molecular regulatory pathways that govern presynaptic function. |
format | Online Article Text |
id | pubmed-8166971 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-81669712021-06-02 An optogenetic method for investigating presynaptic molecular regulation Kay, Yuni Herring, Bruce E. Sci Rep Article While efficient methods are well established for studying postsynaptic protein regulation of glutamatergic synapses in the mammalian central nervous system, similarly efficient methods are lacking for studying proteins regulating presynaptic function. In the present study, we introduce an optical/electrophysiological method for investigating presynaptic molecular regulation. Here, using an optogenetic approach, we selectively stimulate genetically modified presynaptic CA3 pyramidal neurons in the hippocampus and measure optically-induced excitatory postsynaptic currents produced in unmodified postsynaptic CA1 pyramidal neurons. While such use of optogenetics is not novel, previous implementation methods do not allow basic quantification of the changes in synaptic strength produced by genetic manipulations. We find that incorporating simultaneous recordings of fiber volley amplitude provides a control for optical stimulation intensity and, as a result, creates a metric of synaptic efficacy that can be compared across experimental conditions. In the present study, we utilize our new method to demonstrate that inhibition of synaptotagmin 1 expression in CA3 pyramidal neurons leads to a significant reduction in Schaffer collateral synapse function, an effect that is masked with conventional electrical stimulation. Our hope is that this method will expedite our understanding of molecular regulatory pathways that govern presynaptic function. Nature Publishing Group UK 2021-05-31 /pmc/articles/PMC8166971/ /pubmed/34059719 http://dx.doi.org/10.1038/s41598-021-90244-0 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Kay, Yuni Herring, Bruce E. An optogenetic method for investigating presynaptic molecular regulation |
title | An optogenetic method for investigating presynaptic molecular regulation |
title_full | An optogenetic method for investigating presynaptic molecular regulation |
title_fullStr | An optogenetic method for investigating presynaptic molecular regulation |
title_full_unstemmed | An optogenetic method for investigating presynaptic molecular regulation |
title_short | An optogenetic method for investigating presynaptic molecular regulation |
title_sort | optogenetic method for investigating presynaptic molecular regulation |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8166971/ https://www.ncbi.nlm.nih.gov/pubmed/34059719 http://dx.doi.org/10.1038/s41598-021-90244-0 |
work_keys_str_mv | AT kayyuni anoptogeneticmethodforinvestigatingpresynapticmolecularregulation AT herringbrucee anoptogeneticmethodforinvestigatingpresynapticmolecularregulation AT kayyuni optogeneticmethodforinvestigatingpresynapticmolecularregulation AT herringbrucee optogeneticmethodforinvestigatingpresynapticmolecularregulation |