Fig. 3
GFAP expression and VPM thalamic recording in response to optic inhibition of TNC astrocytes. A-B Representative fluorescent image of GFAP expression in the TNC of TN/eNpHR- (A) and TN/eNpHR + (B) rats in response to the optic inhibition of TNC astrocytes. Scale = 100 μm and 20 μm. C Quantification of GFAP immunostaining in TNC of the TN/eNpHR- and TN/eNpHR + groups in the yellow laser on condition, n = 6/group, two-tailed Welch’s t-test, ****p < 0.0001 compared with another group. D Schematic diagram of in vivo optogenetic manipulation of TNC astrocytes with a yellow laser (590 nm) in conjunction with the thalamic recording (E) Coronal section of the VPM thalamus showing the recording site. F Mean firing rate from the VPM thalamus between the TN and sham groups. Two-tailed unpaired t-test with Welch’s correction, n = 16/group, ***P = 0.0002 compared to sham (G) Representative thalamic discharge (lower one presented in frequency, impulse per second) with raw traces (upper) during light-off and yellow laser-on conditions in the TN/eNpHR + group. H Mean firing rate (presented in Hertz) from VPM thalamus of TN/eNpHR + and TN/eNpHR- groups during light off and on conditions. Two-way repeated-measures ANOVA with Sidak’s multiple comparisons test, n = 8/group, *P = 0.0380 compared to light off condition of TN/eNpHR + (I) Mean firing rate (presented in Hertz) from VPM thalamus of sham/eNpHR + and sham/eNpHR- groups during light off and on conditions. Two-way repeated-measures ANOVA with Sidak’s multiple comparisons tests, n = 8/group, not significant, P = 0.4800 compared to the light-off condition of sham/eNpHR + . J Burst frequency (presented in Hertz) comparison between TN (TN/eNpHR + and TN/eNpHR-) and sham (sham/eNpHR + and sham/eNpHR-) groups in response to light stimulation on and off in TNC. Two-way repeated-measures ANOVA with Sidak’s multiple comparisons tests, n = 8/group, **P = 0.0017 compared to light off condition of TN/eNpHR + . Data are presented as the mean ± SD. K Representative burst traces of TN/eNpHR + rat from the VPM thalamus during light-off and light-on condition in TNC astrocytes