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Postsynaptic densities fragment into subcomplexes upon sonication

Postsynaptic density (PSD) fractions were isolated from rat forebrain and sonicated. Pellets from sonicated samples examined by electron microscopy revealed particles with an electron density similar to PSDs that appeared to be fragments of PSDs. Immuno-gold labeling confirmed that some of these con...

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Autores principales: Dosemeci, Ayse, Tao-Cheng, Jung-Hwa, Bakly, Valerie, Reese, Thomas S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6704671/
https://www.ncbi.nlm.nih.gov/pubmed/31439005
http://dx.doi.org/10.1186/s13041-019-0491-y
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author Dosemeci, Ayse
Tao-Cheng, Jung-Hwa
Bakly, Valerie
Reese, Thomas S.
author_facet Dosemeci, Ayse
Tao-Cheng, Jung-Hwa
Bakly, Valerie
Reese, Thomas S.
author_sort Dosemeci, Ayse
collection PubMed
description Postsynaptic density (PSD) fractions were isolated from rat forebrain and sonicated. Pellets from sonicated samples examined by electron microscopy revealed particles with an electron density similar to PSDs that appeared to be fragments of PSDs. Immuno-gold labeling confirmed that some of these contained PSD-95 and/or SynGAP. Biochemical analysis of supernatant and pellet fractions from sonicated samples showed almost complete recovery of several major PSD components (SynGAP, PSD-95, Shank3, Homer and Glutamate receptors) in the pellet, while the supernatant contained known contaminants of PSD fractions, such as glial acidic fibrillary protein and neurofilament protein, as well as actin and α-actinin, indicating susceptibility of these cytoskeletal elements to mechanical disruption. Size distributions of particulate material in control and sonicated samples were clearly different, with particles in the 40–90 nm range observed only in sonicated samples. Fragmentation of the PSD into subcomplexes containing major constituents suggests a patchwork structure consisting of weakly bound modules, that can be readily dissociated from each other through mechanical disruption. Modular organization and weak association between modules would endow the PSD with lateral structural flexibility. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13041-019-0491-y) contains supplementary material, which is available to authorized users.
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spelling pubmed-67046712019-08-22 Postsynaptic densities fragment into subcomplexes upon sonication Dosemeci, Ayse Tao-Cheng, Jung-Hwa Bakly, Valerie Reese, Thomas S. Mol Brain Research Postsynaptic density (PSD) fractions were isolated from rat forebrain and sonicated. Pellets from sonicated samples examined by electron microscopy revealed particles with an electron density similar to PSDs that appeared to be fragments of PSDs. Immuno-gold labeling confirmed that some of these contained PSD-95 and/or SynGAP. Biochemical analysis of supernatant and pellet fractions from sonicated samples showed almost complete recovery of several major PSD components (SynGAP, PSD-95, Shank3, Homer and Glutamate receptors) in the pellet, while the supernatant contained known contaminants of PSD fractions, such as glial acidic fibrillary protein and neurofilament protein, as well as actin and α-actinin, indicating susceptibility of these cytoskeletal elements to mechanical disruption. Size distributions of particulate material in control and sonicated samples were clearly different, with particles in the 40–90 nm range observed only in sonicated samples. Fragmentation of the PSD into subcomplexes containing major constituents suggests a patchwork structure consisting of weakly bound modules, that can be readily dissociated from each other through mechanical disruption. Modular organization and weak association between modules would endow the PSD with lateral structural flexibility. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13041-019-0491-y) contains supplementary material, which is available to authorized users. BioMed Central 2019-08-22 /pmc/articles/PMC6704671/ /pubmed/31439005 http://dx.doi.org/10.1186/s13041-019-0491-y Text en © The Author(s). 2019 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Dosemeci, Ayse
Tao-Cheng, Jung-Hwa
Bakly, Valerie
Reese, Thomas S.
Postsynaptic densities fragment into subcomplexes upon sonication
title Postsynaptic densities fragment into subcomplexes upon sonication
title_full Postsynaptic densities fragment into subcomplexes upon sonication
title_fullStr Postsynaptic densities fragment into subcomplexes upon sonication
title_full_unstemmed Postsynaptic densities fragment into subcomplexes upon sonication
title_short Postsynaptic densities fragment into subcomplexes upon sonication
title_sort postsynaptic densities fragment into subcomplexes upon sonication
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6704671/
https://www.ncbi.nlm.nih.gov/pubmed/31439005
http://dx.doi.org/10.1186/s13041-019-0491-y
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