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Systematic Quantification of Synapses in Primary Neuronal Culture
Most neurological disorders display impaired synaptic connectivity. Hence, modulation of synapse formation may have therapeutic relevance. However, the high density and small size of synapses complicate their quantification. To improve synapse-oriented screens, we analyzed the labeling performance o...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7516133/ https://www.ncbi.nlm.nih.gov/pubmed/33083769 http://dx.doi.org/10.1016/j.isci.2020.101542 |
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author | Verstraelen, Peter Garcia-Diaz Barriga, Gerardo Verschuuren, Marlies Asselbergh, Bob Nuydens, Rony Larsen, Peter H. Timmermans, Jean-Pierre De Vos, Winnok H. |
author_facet | Verstraelen, Peter Garcia-Diaz Barriga, Gerardo Verschuuren, Marlies Asselbergh, Bob Nuydens, Rony Larsen, Peter H. Timmermans, Jean-Pierre De Vos, Winnok H. |
author_sort | Verstraelen, Peter |
collection | PubMed |
description | Most neurological disorders display impaired synaptic connectivity. Hence, modulation of synapse formation may have therapeutic relevance. However, the high density and small size of synapses complicate their quantification. To improve synapse-oriented screens, we analyzed the labeling performance of synapse-targeting antibodies on neuronal cell cultures using segmentation-independent image analysis based on sliding window correlation. When assessing pairwise colocalization, a common readout for mature synapses, overlap was incomplete and confounded by spurious signals. To circumvent this, we implemented a proximity ligation-based approach that only leads to a signal when two markers are sufficiently close. We applied this approach to different marker combinations and demonstrate its utility for detecting synapse density changes in healthy and compromised cultures. Thus, segmentation-independent analysis and exploitation of resident protein proximity increases the sensitivity of synapse quantifications in neuronal cultures and represents a valuable extension to the analytical toolset for in vitro synapse screens. |
format | Online Article Text |
id | pubmed-7516133 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-75161332020-09-28 Systematic Quantification of Synapses in Primary Neuronal Culture Verstraelen, Peter Garcia-Diaz Barriga, Gerardo Verschuuren, Marlies Asselbergh, Bob Nuydens, Rony Larsen, Peter H. Timmermans, Jean-Pierre De Vos, Winnok H. iScience Article Most neurological disorders display impaired synaptic connectivity. Hence, modulation of synapse formation may have therapeutic relevance. However, the high density and small size of synapses complicate their quantification. To improve synapse-oriented screens, we analyzed the labeling performance of synapse-targeting antibodies on neuronal cell cultures using segmentation-independent image analysis based on sliding window correlation. When assessing pairwise colocalization, a common readout for mature synapses, overlap was incomplete and confounded by spurious signals. To circumvent this, we implemented a proximity ligation-based approach that only leads to a signal when two markers are sufficiently close. We applied this approach to different marker combinations and demonstrate its utility for detecting synapse density changes in healthy and compromised cultures. Thus, segmentation-independent analysis and exploitation of resident protein proximity increases the sensitivity of synapse quantifications in neuronal cultures and represents a valuable extension to the analytical toolset for in vitro synapse screens. Elsevier 2020-09-07 /pmc/articles/PMC7516133/ /pubmed/33083769 http://dx.doi.org/10.1016/j.isci.2020.101542 Text en © 2020 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Article Verstraelen, Peter Garcia-Diaz Barriga, Gerardo Verschuuren, Marlies Asselbergh, Bob Nuydens, Rony Larsen, Peter H. Timmermans, Jean-Pierre De Vos, Winnok H. Systematic Quantification of Synapses in Primary Neuronal Culture |
title | Systematic Quantification of Synapses in Primary Neuronal Culture |
title_full | Systematic Quantification of Synapses in Primary Neuronal Culture |
title_fullStr | Systematic Quantification of Synapses in Primary Neuronal Culture |
title_full_unstemmed | Systematic Quantification of Synapses in Primary Neuronal Culture |
title_short | Systematic Quantification of Synapses in Primary Neuronal Culture |
title_sort | systematic quantification of synapses in primary neuronal culture |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7516133/ https://www.ncbi.nlm.nih.gov/pubmed/33083769 http://dx.doi.org/10.1016/j.isci.2020.101542 |
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