Electrophysiological recording: The whole cell patch clamp technique was used for electrophysiological measurement. Patch-pipettes (Corning 7052 glass, AM Systems Inc, Carlsborg, Washington, USA) were pulled to resistances of 1 to 3 МП (Narishige PP83 puller, Tokyo, Japan) and fire-polished to 4 to 6 МП (Narishige MF83 microforge). A predominantly cesium-based internal dialysis solution blocks outward potassium currents in single AVN myocytes . The cesium-based pipette solution contained (in mmol/L) CsCl 113, NaCl 10, K2ATP 5, MgCl2 0.4, HEPES 10 and glucose 5, titrated to apH of 7.1 by adding CsOH. BAPTA 5 mM was also included in this solution to eliminate the intracellular calcium transient and resulting calcium-activated currents .The ‘pipette-to-bath’ liquid junction potential for this solution was less than -3 mV, therefore, no corrections of membrane potential were made. All recordings of membrane currents and action potentials were made with an Axopatch 1D amplifier (Axon Instruments, Foster City, California, USA) and a CV-4 headstage. Normally, 80% to 90% of the electrode series resistance could be compensated. Experimental solutions: A stock solution of flecainide acetate (10 mg/mL, Tambocor, 3М HealthCare, Loughborough, United Kingdom) was kept at 4°C. Aliquots of this solution were added to normal Tyrode’s solution to produce a solution with the final concentration described in the text. Nickel chloride was obtained from Sigma and dissolved in deionized water to give a 0.5 М stock solution, and serial dilutions were made in sodium-free Tyrode’s solution to give a final concentration of 40 |jM. Test solutions were applied to cells with a multibarrelled, warmed solution application device, which could change bulk solution around a single myocyte within 1s . All experiments were performed at temperatures of 35 to 37°C. Your online shopping could cost you less and less time, you just need a reliable pharmacy to buy zoloft online and be sure this one will always live up to your expectations.