Simultaneous visualization of myosin heavy chain isoforms in single muscle sections

Samo Ribarič (Author), Vita Čebašek (Author)

Abstract

We developed a staining protocol that enables simultaneous visualization of myosin heavy chain (MHC) pure and hybrid muscle fiber types in rat skeletal muscle. Up to eight differ-ent muscle fiber types can be visualized in a single section of the rat extensor digitorum longus muscle, which contains all four adult MHC isoforms and shows plasticity during the denervation-reinnervation process. Triple immunofluores-cent staining of MHC-1, MHC-2a and MHC-2b with primary antibodies BA-D5 (isotype IgG2b), SC-71 (isotype IgG1) and BF-F3 (isotype IgM) and with three fluorophore-labeled iso-type-specific secondary antibodies displays different mus-cle fiber types in a merged image of red, green and blue channels, each in its own color. Immunoperoxidase staining with primary antibody 6H1 directed against MHC-2x can be additionally applied on the same tissue section to facilitate the identification of muscle fibers containing MHC-2x. Triple staining can also be used in combination with other staining procedures to derive more information about the number of capillaries or the oxidative potential of muscle fiber types. Simultaneous visualization of multiple fiber types in a single merged image enables economical use of muscle samples and provides simple and rapid identification of all fiber types that are present in rat limb muscles.

Keywords

histokemija;imunofluorescenčni madeži;vrste mišičnih vlaken;histochemistry;immunofluorescent staining;muscle fiber types;

Data

Language:	English
Year of publishing:	2013
Typology:	1.01 - Original Scientific Article
Organization:	UL MF - Faculty of Medicine
UDC:	616-092
COBISS:	30372825
ISSN:	1422-6405
Views:	242
Downloads:	83
Average score:	0 (0 votes)
Metadata:

Other data

Secondary language:	Slovenian
Secondary keywords:	histokemija;imunofluorescenčni madeži;vrste mišičnih vlaken;
Type (COBISS):	Article
Pages:	str. 312-321
Volume:	ǂVol. ǂ197
Issue:	ǂiss. ǂ4
Chronology:	2013
DOI:	10.1159/000345424
ID:	13211147