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Super Resolution Imaging of Genetically Labeled Synapses in Drosophila Brain Tissue

Understanding synaptic connectivity and plasticity within brain circuits and their relationship to learning and behavior is a fundamental quest in neuroscience. Visualizing the fine details of synapses using optical microscopy remains however a major technical challenge. Super resolution microscopy...

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Autores principales: Spühler, Isabelle A., Conley, Gaurasundar M., Scheffold, Frank, Sprecher, Simon G.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4880563/
https://www.ncbi.nlm.nih.gov/pubmed/27303270
http://dx.doi.org/10.3389/fncel.2016.00142
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author Spühler, Isabelle A.
Conley, Gaurasundar M.
Scheffold, Frank
Sprecher, Simon G.
author_facet Spühler, Isabelle A.
Conley, Gaurasundar M.
Scheffold, Frank
Sprecher, Simon G.
author_sort Spühler, Isabelle A.
collection PubMed
description Understanding synaptic connectivity and plasticity within brain circuits and their relationship to learning and behavior is a fundamental quest in neuroscience. Visualizing the fine details of synapses using optical microscopy remains however a major technical challenge. Super resolution microscopy opens the possibility to reveal molecular features of synapses beyond the diffraction limit. With direct stochastic optical reconstruction microscopy, dSTORM, we image synaptic proteins in the brain tissue of the fruit fly, Drosophila melanogaster. Super resolution imaging of brain tissue harbors difficulties due to light scattering and the density of signals. In order to reduce out of focus signal, we take advantage of the genetic tools available in the Drosophila and have fluorescently tagged synaptic proteins expressed in only a small number of neurons. These neurons form synapses within the calyx of the mushroom body, a distinct brain region involved in associative memory formation. Our results show that super resolution microscopy, in combination with genetically labeled synaptic proteins, is a powerful tool to investigate synapses in a quantitative fashion providing an entry point for studies on synaptic plasticity during learning and memory formation.
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spelling pubmed-48805632016-06-14 Super Resolution Imaging of Genetically Labeled Synapses in Drosophila Brain Tissue Spühler, Isabelle A. Conley, Gaurasundar M. Scheffold, Frank Sprecher, Simon G. Front Cell Neurosci Neuroscience Understanding synaptic connectivity and plasticity within brain circuits and their relationship to learning and behavior is a fundamental quest in neuroscience. Visualizing the fine details of synapses using optical microscopy remains however a major technical challenge. Super resolution microscopy opens the possibility to reveal molecular features of synapses beyond the diffraction limit. With direct stochastic optical reconstruction microscopy, dSTORM, we image synaptic proteins in the brain tissue of the fruit fly, Drosophila melanogaster. Super resolution imaging of brain tissue harbors difficulties due to light scattering and the density of signals. In order to reduce out of focus signal, we take advantage of the genetic tools available in the Drosophila and have fluorescently tagged synaptic proteins expressed in only a small number of neurons. These neurons form synapses within the calyx of the mushroom body, a distinct brain region involved in associative memory formation. Our results show that super resolution microscopy, in combination with genetically labeled synaptic proteins, is a powerful tool to investigate synapses in a quantitative fashion providing an entry point for studies on synaptic plasticity during learning and memory formation. Frontiers Media S.A. 2016-05-26 /pmc/articles/PMC4880563/ /pubmed/27303270 http://dx.doi.org/10.3389/fncel.2016.00142 Text en Copyright © 2016 Spühler, Conley, Scheffold and Sprecher. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Neuroscience
Spühler, Isabelle A.
Conley, Gaurasundar M.
Scheffold, Frank
Sprecher, Simon G.
Super Resolution Imaging of Genetically Labeled Synapses in Drosophila Brain Tissue
title Super Resolution Imaging of Genetically Labeled Synapses in Drosophila Brain Tissue
title_full Super Resolution Imaging of Genetically Labeled Synapses in Drosophila Brain Tissue
title_fullStr Super Resolution Imaging of Genetically Labeled Synapses in Drosophila Brain Tissue
title_full_unstemmed Super Resolution Imaging of Genetically Labeled Synapses in Drosophila Brain Tissue
title_short Super Resolution Imaging of Genetically Labeled Synapses in Drosophila Brain Tissue
title_sort super resolution imaging of genetically labeled synapses in drosophila brain tissue
topic Neuroscience
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4880563/
https://www.ncbi.nlm.nih.gov/pubmed/27303270
http://dx.doi.org/10.3389/fncel.2016.00142
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