Cholinergic stimulation of the Na+/K+ adenosine triphosphatase as revealed by microphysiometry.
The activation of a wide range of cellular receptors has been detected previously using a novel instrument, the microphysiometer. In this study microphysiometry was used to monitor the basal and cholinergic-stimulated activity of the Na+/K+ adenosine triphosphatase (ATPase) (the Na+/K+ pump) in the human rhabdomyosarcoma cell line TE671. Manipulations of Na+/K+ ATPase activity with ouabain or removal of extracellular K+ revealed that this ion pump was responsible for 8.8 +/- 0.7% of the total cellular energy utilization by those cells as monitored by the production of acid metabolites. Activation of the pump after a period of inhibition transiently increased the acidification rate above baseline, corresponding to increases in intracellular [Na+] ([Na+]i) occurring while the pump was off. The amplitude of this transient was a function of the total [Na+]i excursion in the absence of pump activity, which in turn depended on the duration of pump inhibition and the Na+ influx rate. Manipulations of the mode of energy metabolism in these cells by changes of the carbon substrate and use of metabolic inhibitors revealed that, unlike some other cells studied, the Na+/K+ ATPase in TE671 cells does not depend on any one mode of metabolism for its adenosine triphosphate source. Stimulation of cholinergic receptors in these cells with carbachol activated the Na+/K+ ATPase via an increase in [Na+]i rather than a direct activation of the ATPase.
Miller
D L
DL
Molecular Devices Corporation, Menlo Park, California 94025.
Olson
J C
JC
Parce
J W
JW
Owicki
J C
JC
D055592:Biophysical Phenomena; D001703:Biophysics; D002217:Carbachol / Q000494:pharmacology; D002460:Cell Line; D002463:Cell Membrane Permeability; D004734:Energy Metabolism; D004789:Enzyme Activation / Q000187:drug effects; D006801:Humans; D010042:Ouabain / Q000494:pharmacology; D011188:Potassium / Q000494:pharmacology; D011199:Potentiometry / Q000295:instrumentation*; D011950:Receptors, Cholinergic / Q000187:drug effects / Q000378:metabolism*; D012964:Sodium / Q000378:metabolism; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism*
D011950:Receptors, Cholinergic; D010042:Ouabain; D002217:Carbachol; D012964:Sodium; D000254:Sodium-Potassium-Exchanging ATPase; D011188:Potassium