Journal: bioRxiv
Article Title: Serotonergic neurons in the dorsal raphe nucleus scale O₂ utilization in response to traumatic stress
doi: 10.64898/2026.01.14.699586
Figure Lengend Snippet: a. Strategy for labeling DRN 5-HT neurons projecting to the VTA and the vCPU. AAV11-DIO-EGFP was bilaterally injected into the vCPU of Sert -Cre mice, and AAV11-DIO-mCherry was injected into both sides of the VTA. b. Images of retrogradely labeled DRN 5-HT neurons. Violet, anti-TPH2 antibody; mCherry⁺ cells, Sert DRN→VTA neurons; EGFP⁺ cells, Sert DRN→vCPU neurons. c. Percentages of three types of DRN 5-HT neurons in the DRN ( n = 7 mice). d. HyPer7 recordings of type III neurons. Strategy for labeling type III neurons and representative confocal image showing that type III neurons mainly locate in the ventral part of the DRN. e. Attack-locked rHF traces of type III neurons in response to a 10-day CSDS. The black lines indicate the onset of attacks. f. Mean attack-locked rHF of type III neurons in SUS and RES mice during a 10-day CSDS paradigm ( n = 6/12 in SUS/RES mice, respectively). g. HyPer7 recordings of Sert DRN→VTA neurons. h. Attack-locked rHF traces of Sert DRN→VTA neurons following a 10-day CSDS. i. Mean attack-locked rHF of type I neurons in SUS and RES mice ( n = 5/8 in SUS/RES mice). j. HyPer7 recordings of Sert DRN→vCPU neurons. k. Attack-locked rHF traces of Sert DRN→vCPU neurons following a 10-day CSDS. l. Mean attack-locked rHF of type II neurons in SUS and RES mice ( n = 6/6 in SUS/RES mice). Scale bars, 100 μm; * p < 0.05 and *** p < 0.001. f, i, l, two-way RM ANOVA with Sidak’s multiple comparisons test. # p < 0.05, # # p < 0.01, and # # # p < 0.001 relative to the basal level (dashed line) by one-sample t -test ( f, i, l ). n.s., not significant. Data are presented as mean ± SEM.
Article Snippet: Free-floating sections were washed in PBS three times, 5 min/wash, permeabilized with 0.3% Triton X-100 in PBS for 30 min, blocked in blocking solution containing 5% normal goat serum and 0.3% Triton X-100 for 2 h at room temperature, and then stained with a rabbit polyclonal anti-TPH2 primary antibody (1:300, NB100-74555; Novus Biologicals) overnight at 4°C.
Techniques: Labeling, Injection
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