Receptor-activated single channels in intact human platelets.
We report "cell-attached" patch clamp studies of intact human platelets which show receptor-activated single channels. Inclusion of ADP in the patch pipette, but not in the bath, resulted in the appearance of inward currents indicative of single channels tightly coupled to the ADP receptors. The channels had a slope conductance of 11 picosiemens at the resting potential. Removal of 1 mM Ca2+ or replacement of chloride by gluconate in the pipette filling solution had little effect on the slope conductance at the resting potential or on the estimated reversed potential. With isotonic BaCl2 in the pipette, ADP evoked single channel currents with a slope conductance of 10 picosiemens. Thus these channels appear to be permeable to monovalent and divalent cations and selective for cations over anions. Addition of 5 mM Ni2+ (which blocks ADP-evoked rapid calcium entry in fura-2-loaded platelets) to the pipette solution blocked ADP-evoked channel activity. These channels may therefore provide an important mechanism for ADP to activate human platelets within a small fraction of a second.
Physiological Laboratory, Cambridge, United Kingdom.
M P
MP
Mahaut-Smith
S O
SO
Sage
T J
TJ
Rink
D000244:Adenosine Diphosphate / Q000494:pharmacology; D001572:Benzofurans; D001792:Blood Platelets / Q000502:physiology*; D002118:Calcium / Q000097:blood*; D004594:Electrophysiology / Q000379:methods; D005456:Fluorescent Dyes; D016257:Fura-2; D005942:Gluconates / Q000494:pharmacology; D006801:Humans; D066298:In Vitro Techniques; D007473:Ion Channels / Q000187:drug effects / Q000502:physiology*; D007700:Kinetics; D011983:Receptors, Purinergic / Q000502:physiology*; D013050:Spectrometry, Fluorescence
D001572:Benzofurans; D005456:Fluorescent Dyes; D005942:Gluconates; D007473:Ion Channels; D011983:Receptors, Purinergic; D000244:Adenosine Diphosphate; C030691:gluconic acid; D002118:Calcium; D016257:Fura-2