Pacemaker function and neural responsiveness of subserosal interstitial cells of Cajal in the mouse colon

Rhythmic action potentials and intercellular Ca²⁺ waves are generated in smooth muscle cells of colonic longitudinal muscles (LSMC). Longitudinal muscle excitability is tuned by input from subserosal ICC (ICC-SS), a population of ICC with previously unknown function. ICC-SS express Ano1 channels and generate spontaneous Ca²⁺ transients in a stochastic manner. Release of Ca²⁺ and activation of Ano1 channels causes depolarization of ICC-SS and LSMC, leading to activation of L-type Ca²⁺ channels, action potentials, intercellular Ca²⁺ waves and contractions in LSMC. Nitrergic neural inputs regulate the Ca²⁺ events in ICC-SS. Pacemaker activity in longitudinal muscle is an emergent property as a result of integrated processes in ICC-SS and LSMC.

Much is known about myogenic mechanisms in circular muscle (CM) in the gastrointestinal tract, although less is known about longitudinal muscle (LM). Two Ca²⁺ signalling behaviours occur in LM: localized intracellular waves not causing contractions and intercellular waves leading to excitation-contraction coupling. An Ano1 channel antagonist inhibited intercellular Ca²⁺ waves and LM contractions. Ano1 channels are expressed by interstitial cells of Cajal (ICC) but not by smooth muscle cells (SMCs). We investigated Ca²⁺ signalling in a novel population of ICC that lies along the subserosal surface of LM (ICC-SS) in mice expressing GCaMP6f in ICC. ICC-SS fired stochastic localized Ca²⁺ transients. Such events have been linked to activation of Ano1 channels in ICC. Ca²⁺ transients in ICC-SS occurred by release from stores most probably via inositol trisphosphate receptors. This activity relied on influx via store-operated Ca²⁺ entry and Orai channels. No voltage-dependent mechanism that synchronized Ca²⁺ transients in a single cell or between cells was found. Nitrergic agonists inhibited Ca²⁺ transients in ICC-SS, and stimulation of intrinsic nerves activated nitrergic responses in ICC-SS. Cessation of stimulation resulted in significant enhancement of Ca²⁺ transients compared to the pre-stimulus activity. No evidence of innervation by excitatory, cholinergic motor neurons was found. Our data suggest that ICC-SS contribute to regulation of LM motor activity. Spontaneous Ca²⁺ transients activate Ano1 channels in ICC-SS. Resulting depolarization conducts to SMCs, depolarizing membrane potential, activating L-type Ca²⁺ channels and initiating contraction. Rhythmic electrical and mechanical behaviours of LM are an emergent property of SMCs and ICC-SS.