Effects of diadenosine polyphosphates on tear secretion in New Zealand white rabbits

Extracellular diadenosine polyphosphates play important signaling functions in a number of physiological responses. Here we show that diadenosine polyphosphates are normal constituents of tear fluid and are potent stimulators of tear secretion through their interaction with P2Y receptors. Diadenosine tetraphosphate (Ap(4)A) and Ap(5)A were found in rabbit tears under basal conditions at concentrations of 2.92 and 0.58 microM, respectively. Single applications of UTP, ATP, and Ap(4)A increased tear secretion to 160 +/- 8% (n = 16) (P < 0.001), 131 +/- 6% (P < 0.05), and 162 +/- 11% (P < 0.05) of placebo values, respectively. Ap(4)A, Ap(5)A, and Ap(6)A, but not Ap(2)A and Ap(3)A, were able to stimulate tear secretion in a dose-dependent manner. Concentration-response studies produced pD(2) values of 5.56 +/- 0.03, 5.75 +/- 0.12, and 5.50 +/- 0.09 for Ap(4)A, Ap(5)A, and Ap(6)A, respectively, with Ap(4)A showing the greatest efficacy. Diadenosine polyphosphates also stimulated P2Y(1) and P2Y(2) receptors expressed in 1321N1 cells with no apparent effect on the other P2Y receptors tested. Nonselective P2 antagonists did not modify the tear secretion induced by UTP or Ap(4)A in rabbit eyes in vivo or in cloned receptors, except for a weak but significant reduction in stimulated tear secretion by reactive blue 2. These results suggest that diadenosine polyphosphates stimulate tear secretion via a P2Y receptor-mediated mechanism. Comparing the effects of diadenosine polyphosphates applied to the rabbit eye and to cloned P2Y receptors, it appears that the P2Y(2) receptor subtype is responsible for the prosecretory effects of these compounds.