Fig. 6: Inhibiting the M1-RN pathway induced anxiety-like behavior.
From: Stress-induced red nucleus attenuation induces anxiety-like behavior and lymph node CCL5 secretion
a Left: injection of AAV1-hSyn-Cre into the M1 and AAV-CaMKII-DIO-GCaMP6 into the RN. Right: a representative confocal image of GCaMP6+ neurons in the RN. b, c Heatmap showing the Ca2+ signals evoked by exploratory behavior in the OFT in RN glutamatergic neurons from control and RS-treated rats. d, e Averaged responses of control and RS-treated rats (black and red line, mean calcium signal during exploratory behavior; gray area and red area, SEM). f Quantification of the change in calcium signals in response to exploratory behavior in control and RS-treated rats (two-tailed unpaired t test, t = 2.429, P = 0.0223, n = 6 rats per group). g Left: injection of AAV1-hSyn-Cre into the M1 and AAV-CaMKII-DIO-hM4Di-mCherry into the RN. Right: a representative confocal image of hM4Di+ neurons in the RN. h Current-evoked action potentials in a representative hM4Di-infected neuron recorded before, during, and after CNO perfusion (10 mM) (n = 6 neurons; two-tailed paired t test, t = 17.33, P < 0.0001; t = 17.89, P < 0.0001, respectively). i Left: locomotion abilities were not affected by inhibiting M1-RN projections (two-tailed unpaired t test, t = 0.4938, P = 0.6291, n = 8 rats per group). Inhibiting M1-RN projections decreased the time spent in the central zone in DIO-hM4Di rats compared to mCherry control rats (two-tailed unpaired t test, t = 4.072, P = 0.0011, n = 8 rats per group). Right: representative tracks of control and hM4Di rats in the OFT. j Both the open arm ratio and frequency decreased significantly in hM4Di rats compared to mCherry control rats (two-tailed unpaired t test, t = 2.181, 2.649, P = 0.0468, 0.0191, n = 8 rats per group). Right: representative tracks of control and hM4Di rats in the EPM. k CCL5 levels in serum were significantly increased after CNO injection (two-tailed paired t test, t = 5.984, P = 0.0010, n = 4 rats). Data are presented as the mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001. Source data are provided as a Source Data file.
