K(+) transport and volume regulatory response by NKCC in resting rat hindlimb skeletal muscle.
This study tested the hypothesis that the NKCC is involved in volume regulation, specifically regulatory volume increase (RVI), in resting skeletal muscle. Neurally and vascularly isolated rat hindlimbs were perfused with a bovine erythrocyte perfusate containing (42)K or (86)Rb as markers of unidirectional K(+) flux across the sarcolemma. Compared to controls, perfusion with 120 microM bumetanide (a specific inhibitor of the NKCC) decreased J(in)K by 15+/-2%, indicating the functional presence of the NKCC. Experiments with ouabain (to block active K(+) transport by the Na,K ATPase) showed that the bumetanide-sensitive component of J(in)K comprised 35% of the total ouabain-sensitive J(in)K. Inhibition of NKCC resulted in a net loss of water by muscle. When hindlimbs were perfused with hypertonic (380 mOsm/L by addition of sucrose) perfusate for 20 min, after initially blocking K(+) channels with 1 mM barium, J(in)K rapidly (2-3 min) increased 2-fold followed by a rapid decline. This rapid, transient increase in J(in)K was abolished with bumetanide, confirming that perfusion with hypertonic perfusate stimulated NKCC activity and RVI. The hypertonic perfusate also resulted in temporally associated decreases in net water uptake by muscle. It is concluded that a functional NKCC is present in mammalian skeletal muscle and that it is involved in cell volume regulation.
Lindinger
Michael I
MI
Dept. of Human Biology and Nutritional Sciences, University of Guelph, Ontario, Canada. mlinding@uoguelph.ca
Hawke
Thomas J
TJ
Lipskie
Shonda L
SL
Schaefer
Hans D
HD
Vickery
Lisa
L
D000818:Animals; D001464:Barium / Q000494:pharmacology; D001692:Biological Transport; D002034:Bumetanide / Q000494:pharmacology; D048429:Cell Size / Q000502:physiology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006614:Hindlimb; D007700:Kinetics; D008297:Male; D018482:Muscle, Skeletal / Q000098:blood supply / Q000378:metabolism*; D009994:Osmolar Concentration; D010042:Ouabain / Q000494:pharmacology; D010477:Perfusion; D011188:Potassium / Q000378:metabolism*; D015221:Potassium Channels / Q000502:physiology; D011194:Potassium Isotopes; D051381:Rats; D017207:Rats, Sprague-Dawley; D012413:Rubidium / Q000737:chemistry; D028021:Sodium-Potassium-Chloride Symporters / Q000378:metabolism / Q000502:physiology*; D000254:Sodium-Potassium-Exchanging ATPase / Q000037:antagonists & inhibitors / Q000378:metabolism; D013395:Sucrose / Q000494:pharmacology
D004791:Enzyme Inhibitors; D015221:Potassium Channels; D011194:Potassium Isotopes; D028021:Sodium-Potassium-Chloride Symporters; D002034:Bumetanide; D001464:Barium; D013395:Sucrose; D010042:Ouabain; D000254:Sodium-Potassium-Exchanging ATPase; D012413:Rubidium; D011188:Potassium