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...

Descripción completa

Detalles Bibliográficos
Autores principales: Sawano, Shoko, Komiya, Yusuke, Ichitsubo, Riho, Ohkawa, Yasuyuki, Nakamura, Mako, Tatsumi, Ryuichi, Ikeuchi, Yoshihide, Mizunoya, Wataru
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