Patch-clamp recordings of membrane currents evoked during natural synaptic activity in sympathetic neurons

• Published in: Neuroscience 1998-11, Vol.87 (2), p.509-517
• Main Authors: Skok, V.I, Farrugia, G, Ermilov, L.G, Miller, S.M, Szurszewski, J.H
• Format: Article
• Language: English
• Subjects: Animals; Biological and medical sciences; Cell Membrane - physiology; Colon - innervation; Colon - physiology; Electric Stimulation; Evoked Potentials - physiology; Excitatory Postsynaptic Potentials - physiology; Fundamental and applied biological sciences. Psychology; Ganglia, Sympathetic - cytology; Ganglia, Sympathetic - physiology; Kinetics; Male; Mesentery - cytology; Mesentery - physiology; Mice; neuronal nicotinic acetylcholine receptor; neuronal nicotinic receptor channel kinetics; neuronal nicotinic receptor subtypes; Neurons - physiology; Patch-Clamp Techniques; Peripheral nervous system. Autonomic nervous system. Neuromuscular transmission. Ganglionic transmission. Electric organ; sympathetic ganglion; Sympathetic Nervous System - cytology; Sympathetic Nervous System - physiology; Synaptic Membranes - physiology; Vertebrates: nervous system and sense organs
• ISSN: 0306-4522; 1873-7544
• DOI: 10.1016/S0306-4522(98)00137-7

Membrane currents elicited by colonic distension and by electrical stimulation of the intermesenteric nerve containing colonic afferent nerve fibres were recorded from neurons of the mouse superior mesenteric ganglion at 20°C with the whole-cell patch-clamp method. Electrically-evoked excitatory postsynaptic currents reversed at −3.5 mV. At membrane holding voltages of −70 mV and −110 mV, the excitatory postsynaptic currents were characterized by a single exponential decay with a mean (±S.E.M.) time-constant of 17.5±1.3 ms and 15.5±2.3 ms, respectively. Colonic distension evoked a series of the excitatory postsynaptic currents which ranged in amplitude from 10 to 700 pA (at a membrane holding voltage of −70 mV). Hexamethonium (100 μM) applied only to the ganglion abolished both electrically- and distension-evoked excitatory postsynaptic currents, suggesting activation of nicotinic acetylcholine receptors. The decay time-course of distension-evoked single excitatory postsynaptic currents was characterized by one, or, less commonly, by two exponentials. The decay time-constant histograms of distension-evoked single excitatory postsynaptic currents exhibited main kinetic components of 8.1±2.3 ms and 8.2±2.5 ms (peak±S.D.) at −70 and −110 mV membrane holding voltages, respectively. Longer time-constants ranging up to 51 ms were also observed. The number of the distension-evoked excitatory postsynaptic currents with a decay time-constant higher than 20 ms, as well as their mean amplitude, were significantly lower at −110 mV than at −70 mV membrane potential levels, in contrast to the currents with a decay time-constant lower than or equal to 20 ms. The results suggest that colonic afferent nerve fibres activate in the mouse superior mesenteric ganglion neurons a few populations of the postsynaptic nicotinic acetylcholine receptors with different channel kinetics, which are characterized by a lack of voltage sensitivity within −70 to −110 mV membrane potential range, except those with comparatively slow channel kinetics, which are possibly blocked by membrane hyperpolarization.