Simultaneous visualization of myosin heavy chain isoforms in single muscle sections

Ribarič, Samo (author), Čebašek, Vita (author)

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.

• Language: English
• Keywords: histochemistry, immunofluorescent staining, muscle fiber types
• Work type: Article
• Typology: 1.01 - Original Scientific Article
• Organization: MF - Faculty of Medicine
• Publication status: Published
• Publication version: Author Accepted Manuscript
• Year: 2013
• Number of pages: Str. 312-321
• Numbering: Vol. 197, iss. 4
• PID: 20.500.12556/RUL-128834
• UDC: 616-092
• ISSN on article: 1422-6405
• DOI: 10.1159/000345424
• COBISS.SI-ID: 30372825
• Publication date in RUL: 05.08.2021

Record is a part of a journal

• Title: Cells tissues organs
• Shortened title: Cells tissues organs
• Publisher: S. Karger
• ISSN: 1422-6405
• COBISS.SI-ID: 1967124

Secondary language

• Language: Slovenian
• Keywords: histokemija, imunofluorescenčni madeži, vrste mišičnih vlaken

Projects

• Funder: ARRS - Slovenian Research Agency
• Project number: P3-0043
• Name: Molekularni mehanizmi razvoja in delovanja skeletne mišice