Fig. 4
From: Neurons in the Nucleus papilio contribute to the control of eye movements during REM sleep
Optogenetic activation of NPCalb terminals in the oculomotor nucleus. a Schematic representation of the experimental set up used to activate the axon terminals of NPCalb neurons in the oculomotor nucleus (3N). The boxed area represents a higher magnification view of the injection of AAV-EF1a-DIO-hChR2(H134R)-YFP in the NPCalb and the location of the optical fibre used for light delivery above the 3N. b Representative image of ChR2-EYFP-positive terminals in the 3N. c Overlay of averaged EOG recordings showing EMs triggered by optogenetic stimulation of NPCalb neuron terminals in the 3N upon 1 s continuous illumination during REM sleep. Grouped data and single animal values are shown in black and grey, respectively. d Comparative Gaussian distribution of EOG signals triggered by optogenetic activation of NPCalb neuron soma (n = 6) or 3N terminals (n = 4) per condition. Gaussian distribution is a fit to maximal deflection of EOG signals. e, f Averaged probabilities e and latencies f of EMs in response to optogenetic stimulations of NPCalb neuron terminals in 3N during wakefulness and REM sleep (n = 4 mice per group; P = 0.43, t = 0.8895 for wake; P = 0.0016, t = 11.098 for REM; df = 3 in both experimental conditions tested). *P < 0.001 significance was calculated using a paired t test. g Summary of the mean duration of wake, NREM and REM sleep bouts, during a 1 h period of light activation delivered every minute of NPCalb cell soma at 1 s continous light (n = 9 and 6, P = 0.79, t = 0.832), 10 Hz (n = 7 and 7, P = 0.69, t = 0.876), and 1 Hz (n = 7 and 5, P = 0.28, t = 0.0.65); df = 35, when comparing ChR2-transduced to control animals. h Summary of the mean duration of wake, NREM and REM sleep bouts during a 1 h period of 1 s continuous stimulation delivered every minute to activate the NPCalb neuron terminals in 3N (n = 4 in each group; P = 0.78, t = 0.8691 for wake; P = 0.941, t = 0.517 for NREM; and P = 0.592, t = 1.167 for REM; df = 18 when comparing ChR2-transduced to control animals). Neither the activation of the NPCalb cell soma nor the activation of their terminals in the 3N had a significant effect on the duration of wakefulness, NREM and REM sleep episodes. Error bars represent SEM values
