Relative distribution of three major lactate transporters in frozen human tissues and their localization in unfixed skeletal muscle

• Published in: Muscle & nerve 2002-07, Vol.26 (1), p.101-112
• Main Authors: Fishbein, William N., Merezhinskaya, Natalya, Foellmer, John W.
• Format: Article
• Language: English
• Subjects: Antibody Specificity; Biological and medical sciences; Blotting, Western; Cryopreservation; Electrophoresis, Polyacrylamide Gel; fiber-type distribution; fiber-type grouping; Fundamental and applied biological sciences. Psychology; Humans; Immune Sera - isolation & purification; Immunohistochemistry; lactate transport; monocarboxylate transport; Monocarboxylic Acid Transporters - biosynthesis; motor unit anatomy; Muscle Fibers, Skeletal - cytology; Muscle Fibers, Skeletal - metabolism; Muscle Proteins; Muscle, Skeletal - cytology; Muscle, Skeletal - metabolism; Organ Specificity; Striated muscle. Tendons; Symporters - biosynthesis; Vertebrates: osteoarticular system, musculoskeletal system
• ISSN: 0148-639X; 1097-4598
• DOI: 10.1002/mus.10168

We have prepared affinity‐purified rabbit polyclonal antibodies to the near‐C‐terminal peptides of human monocarboxylate transporters (MCTs) 1, 2, and 4 coupled to keyhole limpet hemocyanin. Each antiserum reacted only with its specific peptide antigen and gave a distinct molecular weight band (blocked by preincubation with antigen) after chemiluminescence reaction on Western blots from sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS‐PAGE) of tissue membrane proteins. Densitometry showed distinctive expression patterns for each MCT in a panel of 15 frozen human tissues, with the distribution of MCT1 ≫L:MCT2>MCT4. Fluorescence microscopy of unfixed skeletal muscle using fluorescein‐conjugated secondary antibody was correlated with reverse adenosine triphosphatase (ATPase) stained sequential sections to identify fiber‐type localization. MCT1 expression was high in the sarcolemma of type 1 fibers, modest to low in type 2a fibers, and almost absent in type 2b fibers. In contrast, MCT4 expression was low to absent in the membrane of most type 1 fibers, but high in most 2a and in all 2b fibers, favoring the view that their high lactate levels during work may be channeled in part to neighboring type 1 (and perhaps 2a) fibers for oxidation, thereby delaying fatigue. MCT2 expression was limited to the sarcolemma of a type 1 fiber subset, which varied from