The effects of muscarine and adrenaline on patch-clamped frog cardiac parasympathetic neurones

Abstract

1. The whole-cell patch-clamp technique was used to record membrane currents from neurones which were acutely dissociated from the intra-atrial parasympathetic ganglia of Rana pipiens. The effects of muscarine and adrenaline were observed at a holding potential of -30 mV. Extracellular potassium concentration ([K+]o) was 2, 6 or 20 mM. 2. Muscarine (10 microM) produced inward current in thirteen cells, outward current in eighteen cells and seven cells were unaffected. Inward currents were observed in six out of ten neurones in which the intracellular solution contained adenosine triphosphate (ATP; 100 microM) and outward currents were seen in eleven out of fourteen neurones which contained adenosine 3',5'-cyclic monophosphate (cyclic AMP; 100 microM). 3. In five out of nine cells tested, the inward current produced by muscarine was attributable to a 30% depression of a voltage-dependent current which resembled the M-current (IM). Muscarine-induced inward current in the other four cells involved a steady-state conductance increase that reached a null potential at -10 mV. Modest IM suppression also contributed to the response in three of these four cells. 4. Adrenaline (10 or 100 microM) produced inward currents in twelve cells, outward current in ten cells and three cells were unaffected. Outward currents were only seen in cells which contained ATP or cyclic AMP (ten out of sixteen cells) whereas inward currents were seen in eight out of nine cells which did not contain adenosine nucleotides. These inward currents were always attributable to IM suppression. 5. The outward currents induced by muscarine and adrenaline resulted from an increase in a potassium conductance (GK) that exhibited inward rectification.