The relationship between beta-adrenoceptors and adrenergic responsiveness in trout (Oncorhynchus mykiss) and eel (Anguilla rostrata) erythrocytes.
Experiments were performed in vitro specifically to elucidate the underlying mechanism(s) of the attenuated adrenergic responses of eel (Anguilla rostrata) erythrocytes. This was achieved by comparing beta-adrenoceptor numbers and affinities in addition to (i) Na+/H+ exchange activity, (ii) cell swelling and (iii) cyclic AMP formation mediated by catecholamines in eel and trout (Oncorhynchus mykiss) erythrocytes under normoxic and hypoxic conditions. Under normoxic conditions, eel erythrocytes displayed a total absence of Na+/H+ exchange activity (as determined from measurements of extracellular pH) after addition of noradrenaline (50-1000 nmol l-1) in contrast to a pronounced dose-dependent response in trout. Incubation of the blood under hypoxic conditions, to achieve approximately 50% haemoglobin O2-saturation, further increased the extent of Na+/H+ exchange activation in trout and elicited a statistically significant, although physiologically small (10% of the response in trout), activation of H+ extrusion activity in eel. Catecholamine-mediated cell swelling, although obvious in trout, was absent in eel when estimated under hypoxic conditions. Eel erythrocytes possessed approximately 50% fewer surface beta-adrenoceptors than did trout erythrocytes, although the dissociation constants (KD) of these receptors did not differ between eel and trout. The numbers and affinities of the erythrocyte beta-adrenoceptors were not significantly affected by the hypoxic incubation. Both eel and trout erythrocytes displayed a dose-dependent elevation of cyclic AMP concentration in response to noradrenaline that was further increased by hypoxia. Surprisingly, eel erythrocytes produced larger quantities of cyclic AMP despite the lower numbers of surface beta-adrenoceptors. Thus, the absence of adrenergic swelling and the attenuated H+ extrusion response in eel erythrocytes cannot be attributed to insufficient numbers of beta-adrenoceptors or to functional uncoupling of these receptors from adenylate cyclase. Instead, the differences between trout and eel may reflect differing numbers of Na+/H+ exchangers or fundamental differences in the manner by which these exchangers are activated by cyclic AMP.
Perry
S F
SF
Department of Biology, University of Ottawa, Ontario, Canada.
Reid
S D
SD
D000811:Anguilla / Q000097:blood*; D000818:Animals; D002352:Carrier Proteins / Q000097:blood; D000242:Cyclic AMP / Q000097:blood; D004912:Erythrocytes / Q000187:drug effects / Q000378:metabolism*; D000860:Hypoxia / Q000097:blood; D066298:In Vitro Techniques; D009638:Norepinephrine / Q000494:pharmacology; D011943:Receptors, Adrenergic, beta / Q000187:drug effects / Q000378:metabolism*; D012964:Sodium / Q000097:blood; D017923:Sodium-Hydrogen Exchangers; D013045:Species Specificity; D014337:Trout / Q000097:blood*
D002352:Carrier Proteins; D011943:Receptors, Adrenergic, beta; D017923:Sodium-Hydrogen Exchangers; D012964:Sodium; D000242:Cyclic AMP; D009638:Norepinephrine