Electrophysiological and morphological classification of myenteric neurons in the proximal colon of the guinea-pig

Abstract

Intracellular recordings were made from myenteric neurons in the proximal colon of the guinea-pig. The electrical behaviour of the neurons in response to intracellular depolarizing current pulses, and to internodal strand stimulation, was recorded. The intracellular electrode contained the intracellular marker biocytin which was injected into impaled neurons for subsequent histochemistry. Proximal colon myenteric neurons displayed electrophysiological properties similar to myenteric neurons in the small intestine, and were classified as either AH- or S-neurons. AH-neurons were characterized by the presence of a slow afterhyperpolarization following an action potential. Internodal strand stimulation evoked slow excitatory synaptic potentials in five out of six AH-neurons tested, but did not evoke fast excitatory synaptic potentials in 26 AH-neurons tested. S-neurons lacked a slow afterhyperpolarization, but internodal strand stimulation evoked fast excitatory synaptic potentials in all 113 neurons and slow excitatory synaptic potentials in seven out of 17 tested. A subpopulation of AH-neurons displayed a rhythmic oscillation in membrane potential which could be triggered by an action potential. S-neurons could be subdivided into those that fired tonically and those that fired phasically in response to long depolarizing current pulses. About 80% of the AH-neurons were immunoreactive for calbindin, as were 10% of S-neurons. A further 17% of S-neurons, but no AH neurons, were calretinin immunoreactive. Morphological analysis of filled neurons revealed eight distinct classes. Neurons electrophysiologically classified as AH typically had a large, oval soma and several long tapering processes. Processes of AH-neurons branched into many adjacent ganglia. Almost all S-neurons were uniaxonal and many axons ended in an expansion bulb in the myenteric plexus. S-neurons typically had broad, lamellar processes, or short, spiny processes. Roughly equal proportions of S-neurons had oral or anal projections. However, almost all S-neurons that were immunoreactive for calbindin or calretinin projected orally. The results indicate that myenteric neurons in the proximal colon of the guinea-pig are electrophysiologically similar to myenteric neurons in the small intestine, but there are a greater number of morphological and chemical categories.