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Fig. 3. Effects of ACh on ADP-ribosyl cyclase activity and activation of Ca²⁺ oscillations by cADPR. (A) Typical fluorescence signal (ex=300 nm; em=420 nm) of a permeabilized cell induced by 1 µM ACh in control conditions and in presence of methoctramine (10 µM, 3 minutes). (B) Compiled data showing the effects of methoctramine (10 µM), 4DAMP (10 nM), anti-CD38 antibody (1 µg/ml), ZnCl₂ (2 mM) on the amplitude of ACh-induced fluorescent signals and the effects of GTPS (100 µM). Myocytes were superfused with a permeabilization solution (100 µg/ml saponin) containing 100 µM NGD+ and 100 µM ATP. All experiments were made within 10 minutes following application of permeabilization solution. (C) Typical Ca²⁺ signals induced by cADPR (100 nM) in intact or permeabilized cells. (D) Compiled data from permeabilized cells showing the effects of heparin (1 mg/ml, 5 minutes), ryanodine (10 µM, 10 minutes), 8Br-cADPR (50 µM, 5 minutes) and rapamycin (10 µM, 10 minutes) on the amplitude of the first Ca²⁺ peak of cADPR-induced Ca²⁺ responses (left) and the percentage of oscillating cells (right). Myocytes were loaded with 2 µM Fluo4-AM and permeabilized with 10 µg/ml saponin. Data are means ± s.e.m. with the number of tested cells indicated in parentheses (stars indicate P<0.05).

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