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3D imaging of PSD-95 in the mouse brain using the advanced CUBIC method

AIMS: Postsynaptic density – 95 kDa protein (PSD95) is an important molecule on the postsynaptic membrane. It interacts with many other proteins and plays a pivotal role in learning and memory formation. Its distribution in the brain has been studied previously using in situ hybridization as well as...

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Autores principales: Liang, Huazheng, Wang, Hongqin, Wang, Shaoshi, Francis, Richard, Paxinos, George, Huang, Xufeng
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6134724/
https://www.ncbi.nlm.nih.gov/pubmed/30208945
http://dx.doi.org/10.1186/s13041-018-0393-4
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author Liang, Huazheng
Wang, Hongqin
Wang, Shaoshi
Francis, Richard
Paxinos, George
Huang, Xufeng
author_facet Liang, Huazheng
Wang, Hongqin
Wang, Shaoshi
Francis, Richard
Paxinos, George
Huang, Xufeng
author_sort Liang, Huazheng
collection PubMed
description AIMS: Postsynaptic density – 95 kDa protein (PSD95) is an important molecule on the postsynaptic membrane. It interacts with many other proteins and plays a pivotal role in learning and memory formation. Its distribution in the brain has been studied previously using in situ hybridization as well as immunohistochemistry. However, these studies are based on 2 dimensional (2D) sections and results are presented with a few sections. The present study aims to show PSD-95 distribution in 3 dimensions (3D) without slicing the brain tissue of C57BL/6 mice into sections using the advanced CUBIC technique. METHODS: Immunofluorescent staining using a PSD-95 antibody was performed on a half of the mouse brain after clarifying it using the advanced CUBIC protocol. The brain tissue was imaged using a Zeiss Z1 light sheet microscope and 3D reconstruction was completed using the Arivis Vision 4 dimensional (4D) software. RESULTS: The majority of brain nuclei have similar distribution pattern to what has been reported from in situ hybridization and immunohistochemical studies in the mouse. The signal can be easily followed in the 3D and their spatial relationship with adjacent structures clearly demarcated. In the present study, some fiber bundles also showed strong PSD-95 signal, which is different from what was shown in previous studies and need to be confirmed in future studies. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13041-018-0393-4) contains supplementary material, which is available to authorized users.
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spelling pubmed-61347242018-09-13 3D imaging of PSD-95 in the mouse brain using the advanced CUBIC method Liang, Huazheng Wang, Hongqin Wang, Shaoshi Francis, Richard Paxinos, George Huang, Xufeng Mol Brain Micro Report AIMS: Postsynaptic density – 95 kDa protein (PSD95) is an important molecule on the postsynaptic membrane. It interacts with many other proteins and plays a pivotal role in learning and memory formation. Its distribution in the brain has been studied previously using in situ hybridization as well as immunohistochemistry. However, these studies are based on 2 dimensional (2D) sections and results are presented with a few sections. The present study aims to show PSD-95 distribution in 3 dimensions (3D) without slicing the brain tissue of C57BL/6 mice into sections using the advanced CUBIC technique. METHODS: Immunofluorescent staining using a PSD-95 antibody was performed on a half of the mouse brain after clarifying it using the advanced CUBIC protocol. The brain tissue was imaged using a Zeiss Z1 light sheet microscope and 3D reconstruction was completed using the Arivis Vision 4 dimensional (4D) software. RESULTS: The majority of brain nuclei have similar distribution pattern to what has been reported from in situ hybridization and immunohistochemical studies in the mouse. The signal can be easily followed in the 3D and their spatial relationship with adjacent structures clearly demarcated. In the present study, some fiber bundles also showed strong PSD-95 signal, which is different from what was shown in previous studies and need to be confirmed in future studies. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13041-018-0393-4) contains supplementary material, which is available to authorized users. BioMed Central 2018-09-12 /pmc/articles/PMC6134724/ /pubmed/30208945 http://dx.doi.org/10.1186/s13041-018-0393-4 Text en © The Author(s). 2018 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 Micro Report
Liang, Huazheng
Wang, Hongqin
Wang, Shaoshi
Francis, Richard
Paxinos, George
Huang, Xufeng
3D imaging of PSD-95 in the mouse brain using the advanced CUBIC method
title 3D imaging of PSD-95 in the mouse brain using the advanced CUBIC method
title_full 3D imaging of PSD-95 in the mouse brain using the advanced CUBIC method
title_fullStr 3D imaging of PSD-95 in the mouse brain using the advanced CUBIC method
title_full_unstemmed 3D imaging of PSD-95 in the mouse brain using the advanced CUBIC method
title_short 3D imaging of PSD-95 in the mouse brain using the advanced CUBIC method
title_sort 3d imaging of psd-95 in the mouse brain using the advanced cubic method
topic Micro Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6134724/
https://www.ncbi.nlm.nih.gov/pubmed/30208945
http://dx.doi.org/10.1186/s13041-018-0393-4
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