Modulation of tetrodotoxin-resistant Na⁺ channels by amitriptyline in dural afferent neurons

Migraine is characterized by recurrent and disabling headaches; therefore, several drugs have been widely prescribed to prevent acute migraine attacks. Amitriptyline, a tricyclic antidepressant, is among the most commonly administered. It is poorly known, however, whether amitriptyline modulates the excitability of dural afferent neurons that transmit pain signals from the dura mater. In this study, the effects of amitriptyline on tetrodotoxin-resistant (TTX-R) Na⁺ channels were examined in acutely isolated rat dural afferent neurons, which were identified by the fluorescent dye DiI. The TTX-R Na⁺ currents (I_Na) were recorded from medium-sized DiI-positive neurons using a whole-cell patch clamp technique. Amitriptyline (3 μM) slightly reduced the peak component of transient I_Na and induced a marked decrease in the steady-state component of transient TTX-R I_Na, as well as in the slow ramp-induced TTX-R I_Na. Our findings suggest that amitriptyline specifically inhibits persistent Na⁺ currents mediated by TTX-R Na⁺ channels. While amitriptyline had minor effects on voltage-activation/inactivation, it increased the extent of the use-dependent inhibition of TTX-R Na⁺ channels. Amitriptyline also affected the inactivation kinetics of TTX-R Na⁺ channels by significantly accelerating the inactivation of TTX-R Na⁺ channels and slowing the subsequent recovery. Amitriptyline decreased the number of action potentials by increasing the threshold for their generation. In conclusion, the amitriptyline-mediated diverse modulation of TTX-R Na⁺ channels would be, at least in part, responsible for its prophylactic efficacy for migraine attacks.