Additional file 2 of Neurocircuitry underlying the antidepressant effect of retrograde facial botulinum toxin in mice
Creators: name: Ni, Linhui
name: Chen, Hanze
name: Xu, Xinxin
name: Sun, Di
name: Cai, Huaying
name: Wang, Li
name: Tang, Qiwen
name: Hao, Yonggang
name: Cao, Shuxia
name: Hu, Xingyue
Publisher: name: figshare
Year: 2023
Subjects: subject: Biochemistry
subject: Medicine
subject: Microbiology
subject: FOS: Biological sciences
subject: Cell Biology
subject: Molecular Biology
subject: Neuroscience
subject: Physiology
subject: Pharmacology
subject: Biotechnology
subject: Mental Health

description: Additional file 2: Figure S1. Flaccid paralysis of unilateral whisker intrinsic musculature injected with BoNT/A 1 day later. (a) Normal movement of vibrissae controlled by whisker pad of mice injected with saline. (b–d) Dysfunction of vibrissae protracting due to flaccid paralysis of whisker intrinsic musculature induced by three different dosages (3, 10 and 30 U/kg) of BoNT/A injection. Figure S2. Related to Figure 1, Locomotor and anxiety-like behaviors of mice performed in OFT are not affected post-BoNT/A unilateral facial injection. (a) Schematic of grouping and the OFT testing. (b) Mean speed, total distance travelled and duration in the center zone among subgroups of the group receiving BoNT/A injection 6 weeks prior to the restraint end. n = 10 animals from the subgroup of Naïve + Saline, and n = 13 animals from the other four subgroups, respectively. One-way ANOVA followed by Dunnett’s multiple comparisons test comparing each subgroup with the subgroup of CRS mice injected with saline, F (4, 57) = 0.1381, P = 0.9675 in mean speed; F (4, 57) = 0.1559, P = 0.9595 in total distance; F (4, 57) = 1.06, P = 0.3845 in duration in center zone. The P-value of Dunnett’s multiple comparisons: Naïve + Saline vs. CRS + Saline: P = 0.9820, CRS + Saline vs. CRS + 3 U/kg: P = 0.9999, CRS + Saline vs. CRS + 10 U/kg: P = 0.9999, CRS + Saline vs. CRS + 30 U/kg: P = 0.9713 of mean speed; Naïve + Saline vs. CRS + Saline: P = 0.9890, CRS + Saline vs. CRS + 3 U/kg: P = 0.8969, CRS + Saline vs. CRS + 10 U/kg: P = 0.9987, CRS + Saline vs. CRS + 30 U/kg: P = 0.9397 of total distance; Naïve + Saline vs. CRS + Saline: P = 0.8624, CRS + Saline vs. CRS + 3 U/kg: P = 0.3640, CRS + Saline vs. CRS + 10 U/kg: P = 0.5297, CRS + Saline vs. CRS + 30 U/kg: P = 0.1871 of duration in center zone. (c) Subgroup of CRS mice received 30 U/kg of BoNT/A showed improved duration in the center zone of the OFT than the subgroup of CRS mice injected with saline, though the mean speed and total distance travelled among subgroups were in no significant difference of the group pre-injected with BoNT/A 3 weeks prior to the restraint end. n = 10 animals from the subgroup of Naïve + Saline, and n = 15 animals from the other four subgroups, respectively. one-way ANOVA followed by Dunnett’s multiple comparisons test controlling to the subgroup of mice injected with saline, F (4, 65) = 0.9405, P = 0.4463 in mean speed; F (4, 65) = 0.2468, P = 0.9106 in total distance; F (4, 65) = 2.856, P = 0.0304 in duration in center zone. The P-value of Dunnett’s multiple comparisons: Naïve + Saline vs. CRS + Saline: P = 0.3843, CRS + Saline vs. CRS + 3 U/kg: P = 0.4148, CRS + Saline vs. CRS + 10 U/kg: P = 0.5891, CRS + Saline vs. CRS + 30 U/kg: P = 0.9417 of mean speed; Naïve + Saline vs. CRS + Saline: P = 0.9976, CRS + Saline vs. CRS + 3 U/kg: P = 0.7701, CRS + Saline vs. CRS + 10 U/kg: P = 0.9902, CRS + Saline vs. CRS + 30 U/kg: P = 0.9999 of total distance; Naïve + Saline vs. CRS + Saline: P = 0.0113, CRS + Saline vs. CRS + 3 U/kg: P =0.0786, CRS + Saline vs. CRS + 10 U/kg: P = 0.3684, CRS + Saline vs. CRS + 30 U/kg: P = 0.0314 of duration in center zone. (d) Mean speed, total distance travelled and duration in center zone among subgroups of the group with pre-injection of BoNT/A 3 days prior to the restraint end. n = 10 animals from the subgroup of Naïve + Saline, and n = 9 animals from the other four subgroups, respectively. One-way ANOVA followed by Dunnett’s multiple comparisons test controlling to the subgroup of mice injected with saline, F (4, 41) = 0.6347, P = 0.6407 in mean speed; F (4, 41) = 0.4634, P = 0.7621 in total distance; F (4, 41) = 1.496, P = 0.2210 in duration in center zone. The P-value of Dunnett’s multiple comparisons: Naïve + Saline vs. CRS + Saline: P = 0.7785, CRS + Saline vs. CRS + 3 U/kg: P = 0.7375, CRS + Saline vs. CRS + 10 U/kg: P = 0.3648, CRS + Saline vs. CRS + 30 U/kg: P = 0.9535 of mean speed; Naïve + Saline vs. CRS + Saline: P = 0.9878, CRS + Saline vs. CRS + 3 U/kg: P = 0.9985, CRS + Saline vs. CRS + 10 U/kg: P = 0.8151, CRS + Saline vs. CRS + 30 U/kg: P = 0.9991 of total distance; Naïve + Saline vs. CRS + Saline: P = 0.0755, CRS + Saline vs. CRS + 3 U/kg: P = 0.8505, CRS + Saline vs. CRS + 10 U/kg: P = 0.7714, CRS + Saline vs. CRS + 30 U/kg: P = 0.8700 of duration in center zone. *P &lt; 0.05. Figure S3. Related to Figure 2, Absence of BoNT/A-cl.SNAP25197 in trigeminal sensory nuclear complex, second- or higher-order nucleus of WIM. (a–c) Sp5 and Pr5 of trigeminal nuclear complex and some second-order nucleus of WIM. Scale bar, 200 μm. (d) Subnuclei of PAG, dorsal and median raphe nucleus. Right three, magnified view of the left image. Scale bar, 200 μm. (e) Central and basal lateral amygdala. Scale bar, 200 μm. (f) Hippocampus and lateral habenular nucleus. Scale bar, 250 μm mean grid. (g) Medial prefrontal, motor and sensory cortex. Scale bar, 250 μm mean grid. Abbreviation: BLA, Basal lateral amygdala, CPu, Caudate putamen, d/dlPAG, Dorsal/dorsolateral PAG, DRN, Dorsal raphe nucleus, Hipp, Hippocampus, IL infralimbic cortex, IRt, Intermediate reticular nucleus, LHb, Lateral habenular nucleus, l/vlPAG, Lateral/Ventrolateral PAG, M1, Primary motor cortex, M2, Secondary motor cortex, MdD, Dorsal part of medullary reticular nucleus, MRN, Median raphe nucleus, PCRt, Parvicellular reticular nucleus, PrL, Prelimbic cortex, S1, Primary sensory cortex, Sp5C, Caudal part of Sp5, Sp5I, Interpolar part of Sp5, Sp5O, Oral part of Sp5. Figure S4. Related to Figure 3, wFMNs are innervated preferentially by vGluT2 terminals of second-order nucleus. (a) Absence of CTB-488-labeled neurons in the trigeminal sensory nuclear complex or adjacent regions of FN 10 days later of CTB-488-mixed BoNT/A injection into the unilateral WIM. (b–d) Representative image of CTB-488-labeled neurons in lFN 10 days after CTB-488-mixed BoNT/A WIM injection representing wFNMs which were projected by input vGluT2, SerT and vAChT positive terminals, respectively. Scale bar, 20 μm.