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Am J Physiol Cell Physiol 257: C12-C18, 1989;
0363-6143/89 $5.00
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AJP - Cell Physiology, Vol 257, Issue 1 C12-C18, Copyright © 1989 by American Physiological Society

ARTICLES

Regulation of Na+ and K+ contents in rat thymocytes

N. Senn and R. P. Garay
Institut National de la Sante et de la Recherche Medicale U7/CNRS UA 318, Hopital Necker, Paris, France.

A modified nystatin technique allowed the investigation of the initial rate of Na+ efflux as a function of internal Na+ content under steady-state conditions in rat thymocytes. This kinetic study showed that 1) ouabain-sensitive Na+ efflux as a function of internal Na+ can be adjusted by a three-sites kinetic model, with a maximal pump rate of 581 +/- 79 mmol.l cells-1.h-1 and an apparent dissociation constant for internal Na+ of 10.0 +/- 1.0 mmol/l cells (mean +/- SE of 3 experiments), 2) bumetanide-sensitive Na+ efflux was extremely low compared with the pump efflux (approximately 1%), and 3) ouabain- and bumetanide-resistant Na+ efflux was almost a linear function of internal Na+ content (as expected for a Na+ leak). This "all-pump" mechanism of thymocyte Na+ regulation was confirmed by non-steady-state experiments showing that 1) ouabain induced a rapid net Na+ gain and K+ depletion in fresh thymocytes and completely blocked the recovery of normal cation contents in Na+-loaded-K+-depleted thymocytes, and 2) bumetanide was unable to modify thymocyte Na+ and K+ contents. Na+ extrusion by Na+-loaded thymocytes was unaffected by prostaglandin E2, isoproterenol, or platelet-aggregating factor (PAF) and was slightly impaired in the adult spontaneously hypertensive rat of the Okamoto strain (10% lower rate constant for net Na+ extrusion, P less than 0.05). Concerning cell Na+ regulation, our results do not support the concept that rat thymocytes are more representative of vascular cells than enucleated erythrocytes.


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