Riluzole-Sensitive Slowly Inactivating Sodium Current in Rat Suprachiasmatic Nucleus Neurons

Abstract

The persistent (i.e., slowly inactivating) fraction of the Na current (I_Na,P) regulates excitability of CNS neurons. In isolated rat suprachiasmatic nucleus (SCN) neurons with a ramp-type voltage-clamp protocol, we have studied the properties of a robust current that has the general properties of I_Na,P but exhibits a slow inactivation (I_Na,S). The time dependence of the development of the inactivation was also studied by clamping of the membrane potential at different levels: time constants ranging from ∼50 to ∼700 ms, depending on the voltage level, were revealed. The I_Na,S (50–150 pA) was present in both spontaneously active and silent neurons. The neurons exhibited I_Na,S without visible rundown during ∼1-h recordings. I_Na,S had a threshold between –65 and –60 mV and was maximal at about –45 mV. Tetrodotoxin (TTX; 1 μM) completely and reversibly blocked I_Na,S. Riluzole, an effective blocker of I_Na,P, inhibited reversibly I_Na,S with an EC₅₀ of 1–2 μM. Microapplication of 10 μM riluzole during either extracellular or intracellular recording suppressed spontaneous activity in isolated SCN neurons. In the slice preparation, bath application of 20 μM riluzole resulted in decreased firing rate or complete suppression of spontaneous activity in some neurons (9/14) but had no effect on other neurons (5/14). In riluzole-resistant neurons in cell-attached experiments, low-amplitude current spikes were present in 1 μM TTX. We concluded that I_Na,S is ubiquitously expressed by all SCN neurons and that this current is a necessary but not sufficient depolarizing component of the mechanism for spontaneous firing.