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- Author or Editor: Gert Schaart x
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Objective—To investigate whether protein kinase C (PKC) isoforms are expressed in equine skeletal muscle and determine their distribution in various types of fibers by use of immunofluorescence microscopy.
Animals—5 healthy adult Dutch Warmblood horses.
Procedure—In each horse, 2 biopsy specimens were obtained from the vastus lateralis muscle. Cryosections of equine muscle were stained with PKC isoform (α, β1, β2, δ, ξ, or ζ)-specific polyclonal antibodies and examined by use of a fluorescence microscope. Homogenized muscle samples were evaluated via western blot analysis.
Results—The PKC α, β1, β2, δ, ξ, and ζ isoforms were localized within the fibers of equine skeletal muscle. In addition, PKC α and β2 were detected near or in the plasma membrane of muscle cells. For some PKC isoforms, distribution was specific for fiber type. Staining of cell membranes for PKC α was observed predominantly in fibers that reacted positively with myosin heavy chain (MHC)-IIa; PKC δ and ξ staining were more pronounced in MHC-I-positive fibers. In contrast, MHC-I negative fibers contained more PKC ζ than MHC-I-positive fibers. Distribution of PKC β1 was equal among the different fiber types.
Conclusions and Clinical Relevance—Results indicated that PKC isoforms are expressed in equine skeletal muscle in a fiber type-specific manner. Therefore, the involvement of PKC isoforms in signal transduction in equine skeletal muscle might be dependent on fiber type. ( Am J Vet Res 2004; 65:69–73)
Objective—To investigate the expression and localization of glucose transporter 4 (GLUT4) and fatty acid translocase (FAT/CD36) in equine skeletal muscle.
Sample Population—Muscle biopsy specimens obtained from 5 healthy Dutch Warmblood horses.
Procedures—Percutaneous biopsy specimens were obtained from the vastus lateralis, pectoralis descendens, and triceps brachii muscles. Cryosections were stained with combinations of GLUT4 and myosin heavy chain (MHC) specific antibodies or FAT/CD36 and MHC antibodies to assess the fiber specific expression of GLUT4 and FAT/CD36 in equine skeletal muscle via indirect immunofluorescent microscopy.
Results—Immunofluorescent staining revealed that GLUT4 was predominantly expressed in the cytosol of fast type 2B fibers of equine skeletal muscle, although several type 1 fibers in the vastus lateralis muscle were positive for GLUT4. In all muscle fibers examined microscopically, FAT/CD36 was strongly expressed in the sarcolemma and capillaries. Type 1 muscle fibers also expressed small intracellular amounts of FAT/CD36, but no intracellular FAT/CD36 expression was detected in type 2 fibers.
Conclusions and Clinical Relevance—In equine skeletal muscle, GLUT4 and FAT/CD36 are expressed in a fiber type selective manner. ( Am J Vet Res 2004;65:951–956)