Evidence for P2-purinoceptor-mediated uptake of Ca2+ across a fish (Oreochromis mossambicus) intestinal brush border membrane.
We have studied the effect of ATP on Ca2+ uptake in intestinal brush border membrane vesicles (BBMVs) of the teleost tilapia (Oreochromis mossambicus). ATP stimulated Ca2+ uptake 12-fold over the control, with a linear time course. Ionomycin and detergent treatment did not reduce BBMVs' Ca2+ content, indicating the binding of Ca2+ to a membrane component. A rank order of ATP > ADP > AMP was established for the stimulation of Ca2+ uptake. Adenosine, vanadate, adenosine 5'-[alpha, beta-methylene]triphosphate (a P2x purinoceptor agonist) and adenosine 5'-[gamma-thio]triphosphate (a P-type ATPase inhibitor) were without effect. 2-Methylthioadenosine 5'-triphosphate, a P2y purinoceptor agonist, mimicked the stimulation by ATP. As judged from a kinetic comparison, ATP hydrolysis and the stimulation by ATP of Ca2+ uptake were not compatible. The P2 purinoceptor antagonist suramin and the P2y purinoceptor antagonist Reactive Blue-2 inhibited the Ca2+ uptake stimulated by 1 mM ATP (IC50 0.17 mM and 58 microM respectively). We conclude that ATP-stimulated Ca2+ uptake in tilapia intestine is dissociated from ATPase activity and is mediated through a P2 purinoceptor.
Klaren
P H
PH
Department of Animal Physiology, Faculty of Science, University of Nijmegen, The Netherlands.
Wendelaar Bonga
S E
SE
Flik
G
G
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000502:physiology; D000818:Animals; D001692:Biological Transport; D002118:Calcium / Q000378:metabolism*; D005260:Female; D007413:Intestinal Mucosa / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*; D007700:Kinetics; D008297:Male; D008871:Microvilli / Q000201:enzymology / Q000378:metabolism*; D018048:Receptors, Purinergic P2 / Q000502:physiology*; D017210:Tilapia
D018048:Receptors, Purinergic P2; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; D002118:Calcium