Fig. 5: Inhibiting or ablating RN glutamatergic neurons triggered anxiety-like behavior and an increase in CCL5.
From: Stress-induced red nucleus attenuation induces anxiety-like behavior and lymph node CCL5 secretion
a Schematic of AAV-CaMKII-hM4D(Gi)-mCherry injection bilaterally into the RN. Left: confocal image showing CaMKII-hM4D(Gi)-mCherry (red) expression in RN neurons. Right: an enlarged image from the left panel, n = 3. b 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 = 10.99, P = 0.0001; t = 14.26, P < 0.0001, respectively). c Inhibiting RN glutamatergic neurons had no effect on the locomotion abilities of adult rats (two-tailed unpaired t test, t = 1.235, P = 0.2357, n = 17 rats for control and 15 rats for hM4Di) but decreased the time spent in the central zone in the OFT (two-tailed unpaired t test, t = 3.686, P = 0.0022, n = 17 rats for control and 15 rats for hM4Di). d Inhibiting RN glutamatergic neurons led to a downward trend in the open arm ratio and frequency (two-tailed unpaired t test, t = 2.579, P = 0.0150, n = 15–17 rats; t = 1.101, P = 0.2797, n = 17 rats for control and 15 rats for hM4Di). e CCL5 levels in serum were increased in hM4Di rats after CNO injection but not in control rats (two-tailed paired t test, t = 2.484, P = 0.0475; t = 0.2531, P = 0.8087, respectively, n = 4). f Schematic of the bilateral coinjection of AAV-CaMKII-Cre-GFP and AAV-EF1α-DIO-taCasp3-TEVP into the RN. Confocal image showing reduced vGlu1+ signals (red) in the RN, n = 3. g Ablating RN glutamatergic neurons increased the anxiety level of adult rats, which showed less time spent in the central zone in the OFT than controls (two-tailed unpaired t test, t = 2.783, P = 0.0155, n = 9 rats for control and 6 rats for ablate group). h DIO-taCasp3 rats spent less time in the open arms than controls (two-tailed unpaired t test, t = 3.750, P = 0.0024, n = 9 rats for control and 6 rats for ablate group). i Ablating RN glutamatergic neurons led to an increase in CCL5 levels in serum compared to controls (two-tailed unpaired t test, t = 3.757, P = 0.0094, n = 4 rats). j Schematic of AAV-CaMKII-hM3D(Gq)-mCherry injection bilaterally into the RN. Confocal image showing CaMKII-hM3D(Gq)-mCherry (red) expression in RN neurons. k Current-evoked action potentials in a representative hM3Dq-infected neuron recorded before, during, and after CNO perfusion (10 mM) (n = 6 neurons; two-tailed paired t test, t = 15.34, P < 0.0001; t = 13.22, P < 0.0001, respectively). l Activating RN glutamatergic neurons had no effect on the locomotion abilities of adult rats (two-tailed unpaired t test, t = 1.225, P = 0.2396, n = 8–9 rats) but increased the time spent in the central zone in the OFT (two-tailed unpaired t test, t = 3.022, P = 0.0086, n = 9 rats for control and 8 rats for hM3Dq group). m Activating RN glutamatergic neurons led to an increase in the open arm ratio and frequency in the EPM test (two-tailed unpaired t test, t = 2.432, P = 0.0280, n = 8–9 rats; t = 2.431, P = 0.0281, n = 9 rats for control and 8 rats for hM3Dq group). n CCL5 levels in serum were decreased in hM3Dq stress-treated rats after CNO injection but not in RS rats (two-tailed paired t test, t = 3.461, P = 0.0038; t = 0.3152, P = 0.7573, respectively, n = 4). 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.
