Responses of the Na+/H+ exchanger of European flounder red blood cells to hypertonic, beta-adrenergic and acidotic stimuli.
The transport pathways mediating regulatory volume increase (RVI) and beta-adrenergic responses in red cells of the European flounder Platichthys flesus have been investigated. Hypertonic treatment under a low-PO2 atmosphere led to a complete RVI and to a three- to fourfold increase in Na+ influx. The RVI and the activated Na+ influx were blocked by the transport inhibitors amiloride and 4, 4'-diisothiocyanatostilbene-2,2'-disulphonic acid (DIDS), both at a concentration of 10(-4 )mol l-1, and the RVI was abolished in a Na+-free saline, indicating the involvement of a hypertonically induced Na+/H+ exchanger and an accompanying Cl-/HCO3- exchanger. Both the hypertonically induced Na+ influx and the RVI were blocked by oxygenation of shrunk cells. The beta-adrenergic agonist isoproterenol also strongly activated a Na+ influx and caused cell swelling. This response was also inhibited by amiloride and DIDS but was unaffected by oxygenation. Simultaneous application of isoproterenol and hypertonic shrinkage did not lead to additive Na+ influxes, suggesting that both responses were mediated by the same pool of exchangers. Mild cell acidification activated a Na+ influx under iso-osmotic conditions; amiloride caused partial inhibition of this influx, but oxygenation had no effect. Acid-induced and isoproterenol-induced Na+ fluxes were again non-additive. Thus, the Na+/H+ exchanger of flounder red cells is strongly activated by three physiological stimuli: hypertonic shrinkage, beta-adrenergic hormones and cell acidification. Of these responses, only the first is affected by oxygenation, indicating some differentiation of their respective transduction mechanisms. These characteristics contrast with those of the corresponding exchangers from rainbow trout and eel red cells.
Weaver
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Integrative Physiology Research Group, School of Biological Science, University of Liverpool, Derby Building, Liverpool L69 3BX, UK. cossins@liverpool.ac.uk
Kiessling
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Cossins
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