Ryanodine-sensitive stores regulate the excitability of AH neurons in the myenteric plexus of guinea-pig ileum

Myenteric afterhyperpolarizing (AH) neurons are primary afferent neurons wi thin the gastrointestinal tract. Stimulation of the intestinal mucosa evoke s action potentials (AP) that are followed by a slow afterhyperpolarization (AHP(slow)) in the soma. The role of intracellular Ca2+ ([Ca2+](i)) and ry anodine-sensitive Ca2+ stores in modulating the electrical activity of myen teric AH neurons was investigated by recording membrane potential and bis-f ura-2 fluorescence from 34 AH neurons. Mean resting [Ca2+](i) was similar t o 200 nM. Depolarizing current pulses that elicited APs evoked AHP(slow) an d an increase in [Ca2+](i), with similar time courses. The amplitudes and d urations of AHP(slow) and the Ca2+ transient were proportional to the numbe r of evoked APs, with each AP increasing [Ca2+](i) by similar to 50 nM. Rya nodine (10 muM) significantly reduced both the amplitude and duration (by 6 0%) of the evoked Ca2+ transient and AHP(slow) over the range of APs tested (1-15). Calcium-induced calcium release (CICR) was graded and proportional to the number of APs, with each AP triggering a rise in [Ca2+](i) of simil ar to 30 nM Ca2+ via CICR. This indicates that CICR amplifies Ca2+ influx. Similar changes in [Ca2+](i) and AHP(slow) were evoked by two APs in contro l and six APs in ryanodine. Thus, the magnitude of the change in bulk [Ca2](i) and not the source of the Ca2+ is the determinant of the magnitude of AHP(slow). Furthermore, lowering of free [Ca2+](i), either by reducing extr acellular Ca2+ or injecting high concentrations of Ca2+ buffer, induced dep olarization, increased excitability, and abolition of AHP(slow). In additio n, activation of synaptic input to AH neurons elicited a slow excitatory po stsynaptic potential (sEPSP) that was completely blocked in ryanodine. Thes e results demonstrate the importance of [Ca2+](i) and CICR in sensory proce ssing in AH neurons. Activity-dependent CICR may be a mechanism to grade th e output of AH neurons according to the intensity of sensory input.