01 io 100

HCRT concentration (pM)

Figure 2. Hcrt activates LC neurons. A: Bath application of Hcrt-2 activates a typical LC neuron recorded in a slice preparation (1 ^M Hcrt-2 applied beginning at arrow in trace 1; wash begins at arrow in trace 2). Both traces from same LC neuron. B: "Puff" application of Hcrt-2 to an LC neuron from a micropipette (30-second duration of application indicated by the line above the record). C1: Hcrt-2 (1 ^M) in the presence of TTX (1 ^M) generates a small depolarization and initiates spontaneous discharge (apparently Ca2+ spikes; see panel D) 2: Washout of Hcrt-2 in the continued presence of TTX. Both traces from the same LC neuron. D: A spike obtained during Hcrt-2 application in the presence of TTX at a faster sweep speed. Note broad, slow spike waveform typical of Ca2+ spikes compared with a typical Na+ spike found before TTX application. E: Average effects of different concentrations of Hcrt-2 on the spontaneous discharge rate of LC neurons recorded intracellularly in slices. Action potential frequency was significantly increased by all concentrations of Hcrt-2. An increase in spike frequency was seen even in the presence of TTX (1 ^M) when Hcrt-2 was applied. Taken from ref 6.

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