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Membrane Targeted Horseradish Peroxidase as a Marker for Correlative Fluorescence and Electron Microscopy Studies

Synaptic dynamics and reorganization are fundamental features of synaptic plasticity both during synaptic circuit development and in the mature CNS underlying learning, memory, and experience-dependent circuit rearrangements. Combining in vivo time-lapse fluorescence imaging and retrospective electr...

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Detalles Bibliográficos
Autores principales: Li, Jianli, Wang, Yue, Chiu, Shu-Ling, Cline, Hollis T.
Formato: Texto
Lenguaje:English
Publicado: Frontiers Research Foundation 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2831632/
https://www.ncbi.nlm.nih.gov/pubmed/20204144
http://dx.doi.org/10.3389/neuro.04.006.2010
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author Li, Jianli
Wang, Yue
Chiu, Shu-Ling
Cline, Hollis T.
author_facet Li, Jianli
Wang, Yue
Chiu, Shu-Ling
Cline, Hollis T.
author_sort Li, Jianli
collection PubMed
description Synaptic dynamics and reorganization are fundamental features of synaptic plasticity both during synaptic circuit development and in the mature CNS underlying learning, memory, and experience-dependent circuit rearrangements. Combining in vivo time-lapse fluorescence imaging and retrospective electron microscopic analysis provides a powerful technique to decipher the rules governing dynamics of neuronal structure and synaptic connections. Here we have generated a membrane-targeted horseradish peroxidase (mHRP) that allows identification of transfected cells without obscuring the intracellular ultrastructure or organelles and in particular allows identification of synaptic sites using electron microscopy. The expression of mHRP does not affect dendritic arbor growth or dynamics of transfected neurons. Co-expression of EGFP and mHRP was used to study neuronal morphology at both the light and electron microscopic levels. mHRP expression greatly facilitates 3D reconstruction based on serial EM sections. We expect this reagent will be valuable for studying the mechanisms that guide construction of neuronal networks.
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spelling pubmed-28316322010-03-04 Membrane Targeted Horseradish Peroxidase as a Marker for Correlative Fluorescence and Electron Microscopy Studies Li, Jianli Wang, Yue Chiu, Shu-Ling Cline, Hollis T. Front Neural Circuits Neuroscience Synaptic dynamics and reorganization are fundamental features of synaptic plasticity both during synaptic circuit development and in the mature CNS underlying learning, memory, and experience-dependent circuit rearrangements. Combining in vivo time-lapse fluorescence imaging and retrospective electron microscopic analysis provides a powerful technique to decipher the rules governing dynamics of neuronal structure and synaptic connections. Here we have generated a membrane-targeted horseradish peroxidase (mHRP) that allows identification of transfected cells without obscuring the intracellular ultrastructure or organelles and in particular allows identification of synaptic sites using electron microscopy. The expression of mHRP does not affect dendritic arbor growth or dynamics of transfected neurons. Co-expression of EGFP and mHRP was used to study neuronal morphology at both the light and electron microscopic levels. mHRP expression greatly facilitates 3D reconstruction based on serial EM sections. We expect this reagent will be valuable for studying the mechanisms that guide construction of neuronal networks. Frontiers Research Foundation 2010-02-26 /pmc/articles/PMC2831632/ /pubmed/20204144 http://dx.doi.org/10.3389/neuro.04.006.2010 Text en Copyright © 2010 Li, Wang, Chiu and Cline. http://www.frontiersin.org/licenseagreement This is an open-access article subject to an exclusive license agreement between the authors and the Frontiers Research Foundation, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are credited.
spellingShingle Neuroscience
Li, Jianli
Wang, Yue
Chiu, Shu-Ling
Cline, Hollis T.
Membrane Targeted Horseradish Peroxidase as a Marker for Correlative Fluorescence and Electron Microscopy Studies
title Membrane Targeted Horseradish Peroxidase as a Marker for Correlative Fluorescence and Electron Microscopy Studies
title_full Membrane Targeted Horseradish Peroxidase as a Marker for Correlative Fluorescence and Electron Microscopy Studies
title_fullStr Membrane Targeted Horseradish Peroxidase as a Marker for Correlative Fluorescence and Electron Microscopy Studies
title_full_unstemmed Membrane Targeted Horseradish Peroxidase as a Marker for Correlative Fluorescence and Electron Microscopy Studies
title_short Membrane Targeted Horseradish Peroxidase as a Marker for Correlative Fluorescence and Electron Microscopy Studies
title_sort membrane targeted horseradish peroxidase as a marker for correlative fluorescence and electron microscopy studies
topic Neuroscience
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2831632/
https://www.ncbi.nlm.nih.gov/pubmed/20204144
http://dx.doi.org/10.3389/neuro.04.006.2010
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