The effect of a nitric oxide (NO) donor on high-voltage-activated Ca2+ channel currents ( I Ca) was examined using the whole cell patch-clamp technique in L6–S1 dorsal root ganglion (DRG) neurons innervating the urinary bladder. The neurons were labeled by axonal transport of a fluorescent dye, Fast Blue, injected into the bladder wall. Approximately 70% of bladder afferent neurons exhibited tetrodotoxin (TTX)-resistant action potentials (APs), and 93% of these neurons were sensitive to capsaicin, while the remaining neurons had TTX-sensitive spikes and were insensitive to capsaicin. The peak current density of nimodipine-sensitive L-type Ca2+ channels activated by depolarizing pulses (0 mV) from a holding potential of −60 mV was greater in bladder afferent neurons with TTX-resistant APs (39.2 pA/pF) than in bladder afferent neurons with TTX-sensitive APs (28.9 pA/pF), while the current density of ω-conotoxin GVIA-sensitive N-type Ca2+channels was similar (43–45 pA/pF) in both types of neurons. In both types of neurons, the NO donor, S-nitroso- N-acetylpenicillamine (SNAP) (500 μM), reversibly reduced (23.4–26.6%) the amplitude of I Ca elicited by depolarizing pulses to 0 mV from a holding potential of −60 mV. SNAP-induced inhibition of I Ca was reduced by 90% in the presence of ω-conotoxin GVIA but was unaffected in the presence of nimodipine, indicating that NO-induced inhibition of I Ca is mainly confined to N-type Ca2+ channels. Exposure of the neurons for 30 min to 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ, 10 μM), an inhibitor of NO-stimulated guanylyl cyclase, prevented the SNAP-induced reduction in I Ca. Extracellular application of 8-bromo-cGMP (1 mM) mimicked the effects of NO donors by reducing the peak amplitude of I Ca(28.6% of reduction). Action potential configuration and firing frequency during depolarizing current pulses were not altered by the application of SNAP (500 μM) in bladder afferent neurons with TTX-resistant and -sensitive APs. These results indicate that NO acting via a cGMP signaling pathway can modulate N-type Ca2+ channels in DRG neurons innervating the urinary bladder.
Hydrogen sulfide (H2
S) stimulates capsaicin-sensitive primary afferent neurons in the rat urinary bladder
