Determination of the kinetics of Na+/H+ exchange in platelets using the Coulter S-plus cell counter.
Cell swelling, as measured by electronic cell sizing, is a good indicator of the Na+/H+ exchange activation. In this study the kinetic properties of the Na+/H+ exchanger were determined with the aid of the Coulter S-Plus VI D haematological cell counter. Cell swelling was measured in platelets suspended in Na-propionate medium. The rapid entry and intracellular dissociation of propionic acid induces activation of the exchanger, and in turn the uptake of water by osmosis. The fractional volume increase measured by the Coulter S-Plus was dependent on the external Na-concentration, with Km = 86 mmol/l. Saturation was reached at a propionate concentration of 140 mmol/l. Inhibition by amiloride was dose-dependent with Ki = 24 mumol/l. The activity of the exchanger was not modified by ouabain. These data are generally consistent with those published in previous reports, and indicate that automated haematological analysers are appropriate for the study of this aspect of platelet function.
De Caterina
M
M
Department of Biochemistry and Medical Biotechnologies, University of Naples Federico II, Medical School, Italy.
Strazzullo
P
P
Iacone
R
R
Pompeo
F
F
Varriale
V
V
Grimaldi
E
E
Scopacasa
F
F
D000584:Amiloride / Q000494:pharmacology; D001772:Blood Cell Count / Q000295:instrumentation*; D001792:Blood Platelets / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D010042:Ouabain / Q000494:pharmacology; D010976:Platelet Count; D011422:Propionates / Q000494:pharmacology; D012964:Sodium / Q000378:metabolism; D017923:Sodium-Hydrogen Exchangers / Q000097:blood*
D011422:Propionates; D017923:Sodium-Hydrogen Exchangers; D010042:Ouabain; D000584:Amiloride; D012964:Sodium