Journal: The Journal of Headache and Pain

Article Title: ROS/TRPA1/CGRP signaling mediates cortical spreading depression

doi: 10.1186/s10194-019-0978-z

Figure Lengend Snippet: Both ROS and the TRPA1 activation reversed the inhibitory effects of the anti-CGRP antibody on CSD in the mouse brain slice. CSD was induced by 260 mM KCl. There were four groups: anti-IgG antibody at 0.025 μM (i, n = 6) as the control, anti-CGRP antibody at 0.4 μM in the absence (ii, n = 6) or presence of 50 μM of the TRPA1 agonist, AITC (iii, n = 6) or the ROS activator, H 2 O 2 (vi, n = 6). In order to minimize the animal use, data in anti-IgG antibody control group and the anti-CGRP antibody were adopted and transformed from that in Fig. in our recent paper . Representative trace of the 2nd CSD episode in each group are shown in the panel a . The data showed that both ROS and the TRPA1 activation reversed the prolonged CSD latency ( b ), but not magnitude ( c ) under the perfusion of the anti-CGRP antibody. Data were plotted as percentage of their initial levels (1st CSD episode) and indicated as median (range). Mann-Whitney U test, one-tailed, was used for significant analysis between two independent groups. * p < 0.05, ** p < 0.01. Abbreviation: Ab indicates antibody

Article Snippet: Three groups were designed: (i) pretreatment of the anti-TRPA1 antibody with a total 0.8 μg (Alomone Labs, n = 10) 4 days before CSD induction. (ii, iii) Unconjugated rabbit IgG (H + L) (Sangon, D110502, n = 8) with a total 0.8 μg being applied for both the CSD group and the sham group as controls ( n = 7 in each group).

Techniques: Activation Assay, Slice Preparation, Transformation Assay, MANN-WHITNEY, One-tailed Test

Journal: The Journal of Headache and Pain

Article Title: ROS/TRPA1/CGRP signaling mediates cortical spreading depression

doi: 10.1186/s10194-019-0978-z

Figure Lengend Snippet: Effects of the anti-TRPA1 antibody on MDA level (μmol/mg protein) induced by CSD in the ipsilateral cerebral cortex of rat and correlation analysis of each CSD characteristic with levels of cortical ipsilateral MDA between the anti-TRPA1 antibody or anti-IgG antibody groups. a CSD promoted ipsilateral cortical MDA level, which was inhibited by the pretreatment of anti-TRPA1 antibody perfused into the contralateral i.c.v in rats. Mann-Whitney U test, one-tailed, for significance between each group (* p < 0.05, *** p < 0.001). The reduced CSD magnitude ( d ), but not CSD number ( b ) and latency ( c ) positively correlated with a lower MDA level after the anti-TRPA1 antibody perfusion. Red dotted lines indicated positive correlation between CSD magnitude and MDA level

Article Snippet: Three groups were designed: (i) pretreatment of the anti-TRPA1 antibody with a total 0.8 μg (Alomone Labs, n = 10) 4 days before CSD induction. (ii, iii) Unconjugated rabbit IgG (H + L) (Sangon, D110502, n = 8) with a total 0.8 μg being applied for both the CSD group and the sham group as controls ( n = 7 in each group).

Techniques: MANN-WHITNEY, One-tailed Test

Journal: The Journal of Headache and Pain

Article Title: ROS/TRPA1/CGRP signaling mediates cortical spreading depression

doi: 10.1186/s10194-019-0978-z

Figure Lengend Snippet: Effects of the anti-TRPA1 antibody, perfused into the contralateral ventricle, on cortical susceptibility to CSD in rats. a CSD was induced by topical application of 2 M KCl for 30 min onto cerebral cortex with dura intact via the posterior burr hole on the right parietal bone. The ipsilateral anterior hole was used for CSD recording. The anti-TRPA1 antibody (i, n = 10) or anti-IgG antibody (ii, n = 8) was perfused through a cannula implanted in the contralateral ventricle ( i.c.v ) at 4 days prior to CSD induction. In the sham group (iii), the anti-IgG antibody was i.c.v perfused in the absence of KCl application as the control ( n = 7). The whole ipsilateral cerebral cortical tissue was subsequently used for detecting MDA level immediately after the in vivo experiment. b A representative trace showing CSD propagation wave after i.c.v perfusion of the anti-IgG antibody. CSD number, latency (minute) and magnitude (area under the curve of each CSD wave, mV × minute) were used for quantifying the excitation phase of CSD. The effects of the anti-TRPA1 antibody at 0.8 μg on CSD number are shown in panel ( c ), latency in panel ( d ) and magnitude in panel ( e ). All the values shown are median (range). * p < 0.05, Mann-Whitney U test with one-tailed calculation was used for comparison of the anti-IgG antibody and the anti-TRPA1 antibody group

Article Snippet: Three groups were designed: (i) pretreatment of the anti-TRPA1 antibody with a total 0.8 μg (Alomone Labs, n = 10) 4 days before CSD induction. (ii, iii) Unconjugated rabbit IgG (H + L) (Sangon, D110502, n = 8) with a total 0.8 μg being applied for both the CSD group and the sham group as controls ( n = 7 in each group).

Techniques: In Vivo, MANN-WHITNEY, One-tailed Test

Journal: bioRxiv

Article Title: Acute Administration of Oxytocin in the Functional Recovery of Neurocognitive and Social Deficits Following Juvenile Frontal Traumatic Brain Injury

doi: 10.1101/2025.04.30.651517

Figure Lengend Snippet: Effects of IN-OXT on the Oxytocin System Note. Graphs show group means ± SEM. A) IN-OXT increased OXTR levels at the injury site, B) TBI decreased OXT levels at the injury site and in the AON and CP but IN-OXT treatment mitigated this effect, but there was no effect of TBI or IN-OXT on OXT levels in the PVN or SON. **, p <0.01, ***, p <0.001.

Article Snippet: Primary antibodies anti-OXTR (1:100; Alomone Labs #AVR-013; ), anti-OXT (1:10,000, ImmunoStar #20068; ) and anti-Iba1 (microglia; 1:1000; Wako #019-19741; ) were used.

Techniques:

Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

Article Title: Oxytocin produces thermal analgesia via vasopressin-1a receptor by modulating TRPV1 and potassium conductance in the dorsal root ganglion neurons

doi: 10.4196/kjpp.2018.22.2.173

Figure Lengend Snippet: (A) Thermal paw-withdrawal latency was measured and compared before the injection with 30 min and 60 min after the IP injection of vehicle, OT (1 mg/kg) and OT with OTRA (A selective oxytocin receptor antagonist 1 mg/kg), atosiban (an oxytocin and vasopressin receptor antagonist, 1 mg/kg) and V1aRA (a selective vasopressin 1a receptor antagonist, 1 mg/kg) in Hargreaves' test. ** p<0.001, *** p<0.0001, Two-way RM ANOVA followed by posthoc Bonferroni's test. Error bars represent SEM (n=6, each group). (B) Nocifensive responses at 60 min after IP co-injection of antagonists were compared to the OT injection. ** p<0.001, *** p<0.0001, One-way ANOVA followed by posthoc Bonferroni's test. Error bars represent SEM. (n=6, each group). OT, oxytocin; V1aRA, vasopressin-1a receptor antagonist; OTRA, oxytocin receptor antagonist; ns, not significant.

Article Snippet: The primary antibodies were rabbit anti-V1a receptor polyclonal antibody (1:400; Alomone Labs, UK) and guinea pig anti-TRPV1 polyclonal antibody (1:500; Novus biologicals, USA), diluted in PBS containing 0.3% Triton X-100, 1% normal donkey serum (Jackson ImmunoResearch Co, USA), incubated overnight at 4°C then incubated for 1 h with donkey anti-guinea pig IgG conjugated with cyanine dye 3 (Cy3, 1:200; Jackson ImmunoResearch Co, USA) and donkey anti-rabbit IgG conjugated with fluorescein-isothiocyanate (FITC, 1:200; Jackson ImmunoResearch Co, USA) diluted in the same buffer as the primary antibody.

Techniques: Injection

Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

Article Title: Oxytocin produces thermal analgesia via vasopressin-1a receptor by modulating TRPV1 and potassium conductance in the dorsal root ganglion neurons

doi: 10.4196/kjpp.2018.22.2.173

Figure Lengend Snippet: (A) Representative gels show expression patterns of oxytocin binding receptors and TRPV1 from seven individual DRG neurons. Predicted size for selected markers are oxytocin receptor (158 bp), V1a receptor (151 bp), V1b receptor (194 bp), V2 receptor (V2, 153 bp), TRPV1 (330 bp) and GAPDH (268 bp). (B) The percentage of oxytocin binding receptors-positive cells in 54 GAPDH-positive DRG neurons. (C) The percentage of V1a receptor-positive cells within TRPV1 expressing DRG neurons. MM, molecular marker; NC, negative control.

Article Snippet: The primary antibodies were rabbit anti-V1a receptor polyclonal antibody (1:400; Alomone Labs, UK) and guinea pig anti-TRPV1 polyclonal antibody (1:500; Novus biologicals, USA), diluted in PBS containing 0.3% Triton X-100, 1% normal donkey serum (Jackson ImmunoResearch Co, USA), incubated overnight at 4°C then incubated for 1 h with donkey anti-guinea pig IgG conjugated with cyanine dye 3 (Cy3, 1:200; Jackson ImmunoResearch Co, USA) and donkey anti-rabbit IgG conjugated with fluorescein-isothiocyanate (FITC, 1:200; Jackson ImmunoResearch Co, USA) diluted in the same buffer as the primary antibody.

Techniques: Expressing, Binding Assay, Marker, Negative Control

Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

Article Title: Oxytocin produces thermal analgesia via vasopressin-1a receptor by modulating TRPV1 and potassium conductance in the dorsal root ganglion neurons

doi: 10.4196/kjpp.2018.22.2.173

Figure Lengend Snippet: Representative traces of intracellular calcium responses of cultured sensory neurons to 3 repetitive application of (A) 200 nM CAP for 10 s, (B) repetitive CAP application with pre-application of 5 µM of oxytocin for 120 s, (C) CAP application with 5 µM of oxytocin mixed with 20 µM of atosiban, and (D) 5 µM of oxytocin mixed with 5 µM of V1aRA before the second capsaicin application. (E) The proportion of oxytocin responsive cells within the DRG neurons responding to capsaicin in calcium imaging. (F) Summary of normalized ratio of capsaicin responses by without OT (n=21), OT alone (n=70), and OT with its antagonists, ATO (n=66) and V1aRA (n=70), relative to peak amplitude of 1st CAP transient. Before CAP application, 10 µM PMA was mixed with CAP solution. * p<0.05, One-way ANOVA followed by posthoc Bonferroni's test. Error bars represent SEM. CAP, capsaicin; PMA, Phorbol 12-myristate 13-acetate; OT, oxytocin; ATO, atosiban; V1aRA, vasopressin-1a receptor antagonist; ns, not significant.

Article Snippet: The primary antibodies were rabbit anti-V1a receptor polyclonal antibody (1:400; Alomone Labs, UK) and guinea pig anti-TRPV1 polyclonal antibody (1:500; Novus biologicals, USA), diluted in PBS containing 0.3% Triton X-100, 1% normal donkey serum (Jackson ImmunoResearch Co, USA), incubated overnight at 4°C then incubated for 1 h with donkey anti-guinea pig IgG conjugated with cyanine dye 3 (Cy3, 1:200; Jackson ImmunoResearch Co, USA) and donkey anti-rabbit IgG conjugated with fluorescein-isothiocyanate (FITC, 1:200; Jackson ImmunoResearch Co, USA) diluted in the same buffer as the primary antibody.

Techniques: Cell Culture, Imaging

Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

Article Title: Oxytocin produces thermal analgesia via vasopressin-1a receptor by modulating TRPV1 and potassium conductance in the dorsal root ganglion neurons

doi: 10.4196/kjpp.2018.22.2.173

Figure Lengend Snippet: (A, left panel) Representative recording illustrating effect of 5 µM oxytocin on action potential firing. The effect of oxytocin was reversed after 5-min washout. (A, right panel) Oxytocin decreased the mean number of action potential evoked by current injection (n=12). (B) 20 µM of Atosiban (n=9) and (C) 5 µM of V1aRA (n=9) reversed oxytocin-induced reduction of the number of action potentials in small to medium-sized DRG neurons. * p<0.01, *** p<0.0001, repeated measured-ANOVA followed by posthoc Bonferroni's test. Error bars represent SEM. OT, Oxytocin; V1aRA, vasopressin-1a receptor antagonist; ns, not significant.

Article Snippet: The primary antibodies were rabbit anti-V1a receptor polyclonal antibody (1:400; Alomone Labs, UK) and guinea pig anti-TRPV1 polyclonal antibody (1:500; Novus biologicals, USA), diluted in PBS containing 0.3% Triton X-100, 1% normal donkey serum (Jackson ImmunoResearch Co, USA), incubated overnight at 4°C then incubated for 1 h with donkey anti-guinea pig IgG conjugated with cyanine dye 3 (Cy3, 1:200; Jackson ImmunoResearch Co, USA) and donkey anti-rabbit IgG conjugated with fluorescein-isothiocyanate (FITC, 1:200; Jackson ImmunoResearch Co, USA) diluted in the same buffer as the primary antibody.

Techniques: Injection

Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

Article Title: Oxytocin produces thermal analgesia via vasopressin-1a receptor by modulating TRPV1 and potassium conductance in the dorsal root ganglion neurons

doi: 10.4196/kjpp.2018.22.2.173

Figure Lengend Snippet: (A) Depolarizing (from –110 mV to 0 mV) ramp protocol for voltage-clamp experiments (B, left panel) Representative trace showing effects of oxytocin (5 µM) on I-V relationship response to ramp depolarization. (B, middle panel) Representative trace illustrating markedly enhanced a voltage-activated outward current and slightly enhanced a hyperpolarization-activated inward current (B, right panel) Oxytocin increased the mean peak current density at 0 mV by inducing outward potassium current in small to medium-sized DRG neurons (n=20). (C) 20 µM of atosiban (n=24) and (D) 5 µM of V1aRA blocked oxytocin-induced the voltage-activated outward potassium current and the hyperpolarization-activated inward current. TTX (0.5 µM) were treated in solution of all experiments (n=13). * p<0.01, ** p<0.001, repeated measured-ANOVA followed by posthoc Bonferroni's test. Error bars represent SEM. OT, Oxytocin; V1aRA, vasopressin-1a receptor antagonist; ns, not significant.

Article Snippet: The primary antibodies were rabbit anti-V1a receptor polyclonal antibody (1:400; Alomone Labs, UK) and guinea pig anti-TRPV1 polyclonal antibody (1:500; Novus biologicals, USA), diluted in PBS containing 0.3% Triton X-100, 1% normal donkey serum (Jackson ImmunoResearch Co, USA), incubated overnight at 4°C then incubated for 1 h with donkey anti-guinea pig IgG conjugated with cyanine dye 3 (Cy3, 1:200; Jackson ImmunoResearch Co, USA) and donkey anti-rabbit IgG conjugated with fluorescein-isothiocyanate (FITC, 1:200; Jackson ImmunoResearch Co, USA) diluted in the same buffer as the primary antibody.

Techniques: