Cargando…
A One-Step Immunostaining Method to Visualize Rodent Muscle Fiber Type within a Single Specimen
In this study, we present a quadruple immunostaining method for rapid muscle fiber typing of mice and rats using antibodies specific to the adult myosin heavy chain (MyHC) isoforms MyHC1, 2A, 2X, and 2B, which are common marker proteins of distinct muscle fiber types. We developed rat monoclonal ant...
Autores principales: | , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2016
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5096669/ https://www.ncbi.nlm.nih.gov/pubmed/27814384 http://dx.doi.org/10.1371/journal.pone.0166080 |
_version_ | 1782465502580310016 |
---|---|
author | Sawano, Shoko Komiya, Yusuke Ichitsubo, Riho Ohkawa, Yasuyuki Nakamura, Mako Tatsumi, Ryuichi Ikeuchi, Yoshihide Mizunoya, Wataru |
author_facet | Sawano, Shoko Komiya, Yusuke Ichitsubo, Riho Ohkawa, Yasuyuki Nakamura, Mako Tatsumi, Ryuichi Ikeuchi, Yoshihide Mizunoya, Wataru |
author_sort | Sawano, Shoko |
collection | PubMed |
description | In this study, we present a quadruple immunostaining method for rapid muscle fiber typing of mice and rats using antibodies specific to the adult myosin heavy chain (MyHC) isoforms MyHC1, 2A, 2X, and 2B, which are common marker proteins of distinct muscle fiber types. We developed rat monoclonal antibodies specific to each MyHC isoform and conjugated these four antibodies to fluorophores with distinct excitation and emission wavelengths. By mixing the four types of conjugated antibodies, MyHC1, 2A, 2X, and 2B could be distinguished within a single specimen allowing for facile delineation of skeletal muscle fiber types. Furthermore, we could observe hybrid fibers expressing MyHC2X and MyHC2B together in single longitudinal muscle sections from mice and rats, that was not attained in previous techniques. This staining method is expected to be applied to study muscle fiber type transition in response to environmental factors, and to ultimately develop techniques to regulate animal muscle fiber types. |
format | Online Article Text |
id | pubmed-5096669 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-50966692016-11-18 A One-Step Immunostaining Method to Visualize Rodent Muscle Fiber Type within a Single Specimen Sawano, Shoko Komiya, Yusuke Ichitsubo, Riho Ohkawa, Yasuyuki Nakamura, Mako Tatsumi, Ryuichi Ikeuchi, Yoshihide Mizunoya, Wataru PLoS One Research Article In this study, we present a quadruple immunostaining method for rapid muscle fiber typing of mice and rats using antibodies specific to the adult myosin heavy chain (MyHC) isoforms MyHC1, 2A, 2X, and 2B, which are common marker proteins of distinct muscle fiber types. We developed rat monoclonal antibodies specific to each MyHC isoform and conjugated these four antibodies to fluorophores with distinct excitation and emission wavelengths. By mixing the four types of conjugated antibodies, MyHC1, 2A, 2X, and 2B could be distinguished within a single specimen allowing for facile delineation of skeletal muscle fiber types. Furthermore, we could observe hybrid fibers expressing MyHC2X and MyHC2B together in single longitudinal muscle sections from mice and rats, that was not attained in previous techniques. This staining method is expected to be applied to study muscle fiber type transition in response to environmental factors, and to ultimately develop techniques to regulate animal muscle fiber types. Public Library of Science 2016-11-04 /pmc/articles/PMC5096669/ /pubmed/27814384 http://dx.doi.org/10.1371/journal.pone.0166080 Text en © 2016 Sawano 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 Sawano, Shoko Komiya, Yusuke Ichitsubo, Riho Ohkawa, Yasuyuki Nakamura, Mako Tatsumi, Ryuichi Ikeuchi, Yoshihide Mizunoya, Wataru A One-Step Immunostaining Method to Visualize Rodent Muscle Fiber Type within a Single Specimen |
title | A One-Step Immunostaining Method to Visualize Rodent Muscle Fiber Type within a Single Specimen |
title_full | A One-Step Immunostaining Method to Visualize Rodent Muscle Fiber Type within a Single Specimen |
title_fullStr | A One-Step Immunostaining Method to Visualize Rodent Muscle Fiber Type within a Single Specimen |
title_full_unstemmed | A One-Step Immunostaining Method to Visualize Rodent Muscle Fiber Type within a Single Specimen |
title_short | A One-Step Immunostaining Method to Visualize Rodent Muscle Fiber Type within a Single Specimen |
title_sort | one-step immunostaining method to visualize rodent muscle fiber type within a single specimen |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5096669/ https://www.ncbi.nlm.nih.gov/pubmed/27814384 http://dx.doi.org/10.1371/journal.pone.0166080 |
work_keys_str_mv | AT sawanoshoko aonestepimmunostainingmethodtovisualizerodentmusclefibertypewithinasinglespecimen AT komiyayusuke aonestepimmunostainingmethodtovisualizerodentmusclefibertypewithinasinglespecimen AT ichitsuboriho aonestepimmunostainingmethodtovisualizerodentmusclefibertypewithinasinglespecimen AT ohkawayasuyuki aonestepimmunostainingmethodtovisualizerodentmusclefibertypewithinasinglespecimen AT nakamuramako aonestepimmunostainingmethodtovisualizerodentmusclefibertypewithinasinglespecimen AT tatsumiryuichi aonestepimmunostainingmethodtovisualizerodentmusclefibertypewithinasinglespecimen AT ikeuchiyoshihide aonestepimmunostainingmethodtovisualizerodentmusclefibertypewithinasinglespecimen AT mizunoyawataru aonestepimmunostainingmethodtovisualizerodentmusclefibertypewithinasinglespecimen AT sawanoshoko onestepimmunostainingmethodtovisualizerodentmusclefibertypewithinasinglespecimen AT komiyayusuke onestepimmunostainingmethodtovisualizerodentmusclefibertypewithinasinglespecimen AT ichitsuboriho onestepimmunostainingmethodtovisualizerodentmusclefibertypewithinasinglespecimen AT ohkawayasuyuki onestepimmunostainingmethodtovisualizerodentmusclefibertypewithinasinglespecimen AT nakamuramako onestepimmunostainingmethodtovisualizerodentmusclefibertypewithinasinglespecimen AT tatsumiryuichi onestepimmunostainingmethodtovisualizerodentmusclefibertypewithinasinglespecimen AT ikeuchiyoshihide onestepimmunostainingmethodtovisualizerodentmusclefibertypewithinasinglespecimen AT mizunoyawataru onestepimmunostainingmethodtovisualizerodentmusclefibertypewithinasinglespecimen |