Hydrogen sulfide increases excitability through suppression of sustained potassium channel currents of rat trigeminal ganglion neurons

BACKGROUND: Hydrogen sulfide (H(2)S), an endogenous gaseotransmitter/modulator, is becoming appreciated that it may be involved in a wide variety of processes including inflammation and nociception. However, the role and mechanism for H(2)S in nociceptive processing in trigeminal ganglion (TG) neuron remains unknown. The aim of this study is to investigate distribution of endogenous H(2)S synthesizing enzyme cystathionine-β-synthetase (CBS) expression and role of H(2)S on excitability and voltage-gated potassium channels of TG neurons. METHODS: Immunofluorescence studies were carried out to determine whether CBS was co-expressed in Kv1.1 or Kv1.4-positive TG neurons. Whole cell patch clamp recordings were employed on acutely isolated TG neurons from adult male Sprague Dawley rats (6–8 week old). von Frey filaments were used to examine the pain behavioral responses in rats following injection of sodium hydrosulfide. RESULTS: In rat TG, 77.3±6.6% neurons were immunoreactive for CBS, 85.1±3.8% for Kv1.1 and 97.8±1.1% for Kv1.4. Double staining showed that all CBS labeled cells were Kv1.1 and Kv1.4 positive, but only 92.2±6.1% of Kv1.1 and 78.2±9.9% of Kv1.4 positive cells contained CBS. Application of H(2)S donor NaHS (250 μM) led to a significant depolarization of resting membrane potential recorded from TG neurons. NaHS application also resulted in a dramatic reduction in rheobase, hyperpolarization of action potential threshold, and a significant increase in the number of action potentials evoked at 2X and 3X rheobase stimulation. Under voltage-clamp conditions, TG neurons exhibited transient A-type (I(A)) and sustained outward rectifier K(+) currents (I(K)). Application of NaHS did suppress I(K) density while did not change I(A) density of TG neurons (n=6). Furthermore, NaHS, a donor of hydrogen sulfide, produced a significant reduction in escape threshold in a dose dependent manner. CONCLUSION: These data suggest that endogenous H(2)S generating enzyme CBS was co-localized well with Kv1.1 and Kv1.4 in TG neurons and that H(2)S produces the mechanic pain and increases neuronal excitability, which might be largely mediated by suppressing I(K) density, thus identifying for the first time a specific molecular mechanism underlying pain and sensitization in TG.