Immunohistochemical analysis of MCT1, MCT2 and MCT4 expression in rat plantaris muscle

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Abstract
We addressed the need for histological assessment of myocellular domains occupied by monocarboxylate transporters (MCT1, MCT2 and MCT4). From the perspective of lactate shuttle hypotheses we posited that MCT1 would be highly expressed in oxidative fibres, whereas MCT4 would be found in highly glycolytic fibres. Furthermore, we hypothesized that MCT1 would be detected at interfibrillar as well as at subsarcolemmal and sarcolemmal cell domains, whereas MCT2 and MCT4 abundances would be most prominent at the sarcolemma. To test these hypotheses, we examined cellular locations of MCT1, MCT2 and MCT4 transporter proteins in different fibre types (slow oxidative, SO; fast oxidative glycolytic, FOG; fast glycolytic, FG) in rat plantaris muscles by the avidin–biotin complex (ABC) as well as other methods. The plantaris was used as it is a mixed fibre skeletal muscle. MCTs, glucose transporter (GLUT4) protein, and mitochondrial constituent cytochrome oxidase (COX) abundances were assessed by immunohistochemistry and Western blotting using affinity-purified antibodies. The staining method was specific and stable, which allowed for semiquantitative assessment of MCT expression. As well, confocal laser scanning microscopy assessed MCT isoform localizations. Findings of the present study were: (1) MCT1 is located at the sarcolemma and throughout the cell interior in SO and FOG fibres where the mitochondrial reticulum was present; (2) in contrast, MCT4 was highly expressed in the sarcolemmal domain of FG and FOG fibres but poorly expressed in SO fibres; and (3) confocal laser-scanning microscopy demonstrated that MCT1 and COX are co-localised at both interfibrillar and subsarcolemmal cell domains, whereas MCT2 is only faintly detected at the sarcolemma of oxidative fibres. MCTs and associated proteins are positioned to facilitate the function of the lactate shuttles.