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Capillary pericytes express α-smooth muscle actin, which requires prevention of filamentous-actin depolymerization for detection
Recent evidence suggests that capillary pericytes are contractile and play a crucial role in the regulation of microcirculation. However, failure to detect components of the contractile apparatus in capillary pericytes, most notably α-smooth muscle actin (α-SMA), has questioned these findings. Using...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
eLife Sciences Publications, Ltd
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5862523/ https://www.ncbi.nlm.nih.gov/pubmed/29561727 http://dx.doi.org/10.7554/eLife.34861 |
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author | Alarcon-Martinez, Luis Yilmaz-Ozcan, Sinem Yemisci, Muge Schallek, Jesse Kılıç, Kıvılcım Can, Alp Di Polo, Adriana Dalkara, Turgay |
author_facet | Alarcon-Martinez, Luis Yilmaz-Ozcan, Sinem Yemisci, Muge Schallek, Jesse Kılıç, Kıvılcım Can, Alp Di Polo, Adriana Dalkara, Turgay |
author_sort | Alarcon-Martinez, Luis |
collection | PubMed |
description | Recent evidence suggests that capillary pericytes are contractile and play a crucial role in the regulation of microcirculation. However, failure to detect components of the contractile apparatus in capillary pericytes, most notably α-smooth muscle actin (α-SMA), has questioned these findings. Using strategies that allow rapid filamentous-actin (F-actin) fixation (i.e. snap freeze fixation with methanol at −20°C) or prevent F-actin depolymerization (i.e. with F-actin stabilizing agents), we demonstrate that pericytes on mouse retinal capillaries, including those in intermediate and deeper plexus, express α-SMA. Junctional pericytes were more frequently α-SMA-positive relative to pericytes on linear capillary segments. Intravitreal administration of short interfering RNA (α-SMA-siRNA) suppressed α-SMA expression preferentially in high order branch capillary pericytes, confirming the existence of a smaller pool of α-SMA in distal capillary pericytes that is quickly lost by depolymerization. We conclude that capillary pericytes do express α-SMA, which rapidly depolymerizes during tissue fixation thus evading detection by immunolabeling. |
format | Online Article Text |
id | pubmed-5862523 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | eLife Sciences Publications, Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-58625232018-03-22 Capillary pericytes express α-smooth muscle actin, which requires prevention of filamentous-actin depolymerization for detection Alarcon-Martinez, Luis Yilmaz-Ozcan, Sinem Yemisci, Muge Schallek, Jesse Kılıç, Kıvılcım Can, Alp Di Polo, Adriana Dalkara, Turgay eLife Neuroscience Recent evidence suggests that capillary pericytes are contractile and play a crucial role in the regulation of microcirculation. However, failure to detect components of the contractile apparatus in capillary pericytes, most notably α-smooth muscle actin (α-SMA), has questioned these findings. Using strategies that allow rapid filamentous-actin (F-actin) fixation (i.e. snap freeze fixation with methanol at −20°C) or prevent F-actin depolymerization (i.e. with F-actin stabilizing agents), we demonstrate that pericytes on mouse retinal capillaries, including those in intermediate and deeper plexus, express α-SMA. Junctional pericytes were more frequently α-SMA-positive relative to pericytes on linear capillary segments. Intravitreal administration of short interfering RNA (α-SMA-siRNA) suppressed α-SMA expression preferentially in high order branch capillary pericytes, confirming the existence of a smaller pool of α-SMA in distal capillary pericytes that is quickly lost by depolymerization. We conclude that capillary pericytes do express α-SMA, which rapidly depolymerizes during tissue fixation thus evading detection by immunolabeling. eLife Sciences Publications, Ltd 2018-03-21 /pmc/articles/PMC5862523/ /pubmed/29561727 http://dx.doi.org/10.7554/eLife.34861 Text en © 2018, Alarcon-Martinez et al http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited. |
spellingShingle | Neuroscience Alarcon-Martinez, Luis Yilmaz-Ozcan, Sinem Yemisci, Muge Schallek, Jesse Kılıç, Kıvılcım Can, Alp Di Polo, Adriana Dalkara, Turgay Capillary pericytes express α-smooth muscle actin, which requires prevention of filamentous-actin depolymerization for detection |
title | Capillary pericytes express α-smooth muscle actin, which requires prevention of filamentous-actin depolymerization for detection |
title_full | Capillary pericytes express α-smooth muscle actin, which requires prevention of filamentous-actin depolymerization for detection |
title_fullStr | Capillary pericytes express α-smooth muscle actin, which requires prevention of filamentous-actin depolymerization for detection |
title_full_unstemmed | Capillary pericytes express α-smooth muscle actin, which requires prevention of filamentous-actin depolymerization for detection |
title_short | Capillary pericytes express α-smooth muscle actin, which requires prevention of filamentous-actin depolymerization for detection |
title_sort | capillary pericytes express α-smooth muscle actin, which requires prevention of filamentous-actin depolymerization for detection |
topic | Neuroscience |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5862523/ https://www.ncbi.nlm.nih.gov/pubmed/29561727 http://dx.doi.org/10.7554/eLife.34861 |
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