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Functional characterisation of filamentous actin probe expression in neuronal cells
Genetically encoded filamentous actin probes, Lifeact, Utrophin and F-tractin, are used as tools to label the actin cytoskeleton. Recent evidence in several different cell types indicates that these probes can cause changes in filamentous actin dynamics, altering cell morphology and function. Althou...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5690639/ https://www.ncbi.nlm.nih.gov/pubmed/29145435 http://dx.doi.org/10.1371/journal.pone.0187979 |
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author | Patel, Shrujna Fok, Sandra Y. Y. Stefen, Holly Tomanić, Tamara Parić, Esmeralda Herold, Rosanna Brettle, Merryn Djordjevic, Aleksandra Fath, Thomas |
author_facet | Patel, Shrujna Fok, Sandra Y. Y. Stefen, Holly Tomanić, Tamara Parić, Esmeralda Herold, Rosanna Brettle, Merryn Djordjevic, Aleksandra Fath, Thomas |
author_sort | Patel, Shrujna |
collection | PubMed |
description | Genetically encoded filamentous actin probes, Lifeact, Utrophin and F-tractin, are used as tools to label the actin cytoskeleton. Recent evidence in several different cell types indicates that these probes can cause changes in filamentous actin dynamics, altering cell morphology and function. Although these probes are commonly used to visualise actin dynamics in neurons, their effects on axonal and dendritic morphology has not been systematically characterised. In this study, we quantitatively analysed the effect of Lifeact, Utrophin and F-tractin on neuronal morphogenesis in primary hippocampal neurons. Our data show that the expression of actin-tracking probes significantly impacts on axonal and dendrite growth these neurons. Lifeact-GFP expression, under the control of a pBABE promoter, caused a significant decrease in total axon length, while another Lifeact-GFP expression, under the control of a CAG promoter, decreased the length and complexity of dendritic trees. Utr261-EGFP resulted in increased dendritic branching but Utr230-EGFP only accumulated in cell soma, without labelling any neurites. Lifeact-7-mEGFP and F-tractin-EGFP in a pEGFP-C1 vector, under the control of a CMV promoter, caused only minor changes in neuronal morphology as detected by Sholl analysis. The results of this study demonstrate the effects that filamentous actin tracking probes can have on the axonal and dendritic compartments of neuronal cells and emphasise the care that must be taken when interpreting data from experiments using these probes. |
format | Online Article Text |
id | pubmed-5690639 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-56906392017-11-30 Functional characterisation of filamentous actin probe expression in neuronal cells Patel, Shrujna Fok, Sandra Y. Y. Stefen, Holly Tomanić, Tamara Parić, Esmeralda Herold, Rosanna Brettle, Merryn Djordjevic, Aleksandra Fath, Thomas PLoS One Research Article Genetically encoded filamentous actin probes, Lifeact, Utrophin and F-tractin, are used as tools to label the actin cytoskeleton. Recent evidence in several different cell types indicates that these probes can cause changes in filamentous actin dynamics, altering cell morphology and function. Although these probes are commonly used to visualise actin dynamics in neurons, their effects on axonal and dendritic morphology has not been systematically characterised. In this study, we quantitatively analysed the effect of Lifeact, Utrophin and F-tractin on neuronal morphogenesis in primary hippocampal neurons. Our data show that the expression of actin-tracking probes significantly impacts on axonal and dendrite growth these neurons. Lifeact-GFP expression, under the control of a pBABE promoter, caused a significant decrease in total axon length, while another Lifeact-GFP expression, under the control of a CAG promoter, decreased the length and complexity of dendritic trees. Utr261-EGFP resulted in increased dendritic branching but Utr230-EGFP only accumulated in cell soma, without labelling any neurites. Lifeact-7-mEGFP and F-tractin-EGFP in a pEGFP-C1 vector, under the control of a CMV promoter, caused only minor changes in neuronal morphology as detected by Sholl analysis. The results of this study demonstrate the effects that filamentous actin tracking probes can have on the axonal and dendritic compartments of neuronal cells and emphasise the care that must be taken when interpreting data from experiments using these probes. Public Library of Science 2017-11-16 /pmc/articles/PMC5690639/ /pubmed/29145435 http://dx.doi.org/10.1371/journal.pone.0187979 Text en © 2017 Patel et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Patel, Shrujna Fok, Sandra Y. Y. Stefen, Holly Tomanić, Tamara Parić, Esmeralda Herold, Rosanna Brettle, Merryn Djordjevic, Aleksandra Fath, Thomas Functional characterisation of filamentous actin probe expression in neuronal cells |
title | Functional characterisation of filamentous actin probe expression in neuronal cells |
title_full | Functional characterisation of filamentous actin probe expression in neuronal cells |
title_fullStr | Functional characterisation of filamentous actin probe expression in neuronal cells |
title_full_unstemmed | Functional characterisation of filamentous actin probe expression in neuronal cells |
title_short | Functional characterisation of filamentous actin probe expression in neuronal cells |
title_sort | functional characterisation of filamentous actin probe expression in neuronal cells |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5690639/ https://www.ncbi.nlm.nih.gov/pubmed/29145435 http://dx.doi.org/10.1371/journal.pone.0187979 |
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