title: Na+-K+ pumps in the transverse tubular system of skeletal muscle fibers preferentially use ATP from glycolysis.
abstract: The Na(+)-K(+) pumps in the transverse tubular (T) system of a muscle fiber play a vital role keeping K(+) concentration in the T-system sufficiently low during activity to prevent chronic depolarization and consequent loss of excitability. These Na(+)-K(+) pumps are located in the triad junction, the key transduction zone controlling excitation-contraction (EC) coupling, a region rich in glycolytic enzymes and likely having high localized ATP usage and limited substrate diffusion. This study examined whether Na(+)-K(+) pump function is dependent on ATP derived via the glycolytic pathway locally within the triad region. Single fibers from rat fast-twitch muscle were mechanically skinned, sealing off the T-system but retaining normal EC coupling. Intracellular composition was set by the bathing solution and action potentials (APs) triggered in the T-system, eliciting intracellular Ca(2+) release and twitch and tetanic force responses. Conditions were selected such that increased Na(+)-K(+) pump function could be detected from the consequent increase in T-system polarization and resultant faster rate of AP repriming. Na(+)-K(+) pump function was not adequately supported by maintaining cytoplasmic ATP concentration at its normal resting level ( approximately 8 mM), even with 10 or 40 mM creatine phosphate present. Addition of as little as 1 mM phospho(enol)pyruvate resulted in a marked increase in Na(+)-K(+) pump function, supported by endogenous pyruvate kinase bound within the triad. These results demonstrate that the triad junction is a highly restricted microenvironment, where glycolytic resynthesis of ATP is critical to meet the high demand of the Na(+)-K(+) pump and maintain muscle excitability.
authors.affiliation: Dept. of Zoology, La Trobe Univ., Melbourne 3086, Victoria, Australia. t.dutka@latrobe.edu.au
authors.forename: T L
authors.initials: TL
authors.lastname: Dutka
authors.forename: G D
authors.initials: GD
authors.lastname: Lamb
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D002712:Chlorides / Q000494:pharmacology; D004734:Energy Metabolism / Q000187:drug effects / Q000502:physiology; D005905:Glyburide / Q000494:pharmacology; D006019:Glycolysis / Q000187:drug effects / Q000502:physiology*; D007004:Hypoglycemic Agents / Q000494:pharmacology; D008297:Male; D009119:Muscle Contraction / Q000187:drug effects / Q000502:physiology; D018763:Muscle Fatigue / Q000502:physiology; D018485:Muscle Fibers, Skeletal / Q000378:metabolism* / Q000648:ultrastructure; D018482:Muscle, Skeletal / Q000166:cytology / Q000378:metabolism*; D010728:Phosphoenolpyruvate / Q000494:pharmacology; D011770:Pyruvate Kinase / Q000494:pharmacology; D051381:Rats; D020318:Rats, Long-Evans; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism*
chemical_list: D002712:Chlorides; D007004:Hypoglycemic Agents; D010728:Phosphoenolpyruvate; D000255:Adenosine Triphosphate; D011770:Pyruvate Kinase; D000254:Sodium-Potassium-Exchanging ATPase; D005905:Glyburide