Journal: European Journal of Pain (London, England)

Article Title: Bidirectional Roles of TRPV1 in a Latent Sensitization Model of Myofascial Low Back Pain

doi: 10.1002/ejp.70255

Figure Lengend Snippet: Repeated NGF injections enhance presynaptic TRPV1 expression in superficial laminae of the ipsilateral lumbar spinal cord. (A) Representative image of the mean fluorescence intensity of TRPV1 in the ipsilateral dorsal horn of the lumbar spinal cord segment L2. Images were acquired under identical conditions, with parameters optimized for NGF‐treated tissue; thus, basal TRPV1 expression in PBS controls may appear faint. (B) TRPV1 is time‐dependently upregulated in spinal dorsal horn after NGF injection (16 bit, scale: 0–65536). Mean ± SEM, n = 5 each, two‐way ANOVA. ns = not significant, p ≥ 0.05; * p < 0.05; # strong effect, Cohen's d ≥ 1. Scale bar: 200 μm. Time points shown are day 1 after the first injection (d1), day 5 after the first injection (d5), day 1 after the second injection (d1′), and day 5 after the second injection (d5′).

Article Snippet: TRPV1 (rabbit, 1:500; Cat. No. ACC‐030, Alomone Labs, Jerusalem, Israel) and NeuN (guinea pig, 1:1000; Cat. No. 266004, Synaptic Systems, Göttingen, Germany) primary antibodies were used to determine TRPV1 expression.

Techniques: Expressing, Fluorescence, Injection

Journal: European Journal of Pain (London, England)

Article Title: Bidirectional Roles of TRPV1 in a Latent Sensitization Model of Myofascial Low Back Pain

doi: 10.1002/ejp.70255

Figure Lengend Snippet: Absence of capsaicin‐responsive DRG neurons in TRPV1 −/− rats. (A) Example of false colour‐coded ratiometric calcium imaging pictures exhibiting the change in intracellular calcium in response to chemical stimulation (capsaicin, potassium): Upper row: TRPV1 −/− rat, lower row: WT rat. Scale bar: 100 μm. (B) Calcium responses of DRG neurons after capsaicin application. Left: Proportion of capsaicin‐responsive DRG neurons vs capsaicin‐unresponsive but potassium‐responsive neurons; middle: Relative change of intracellular calcium in response to capsaicin application; black line: Mean; dotted red line: 23%, limit above which a calcium increase was considered to be a response to capsaicin. (C) TRPV1 −/− rats required higher laser intensities (mW) to evoke nocifensive responses compared to WT, regardless of sex. Thresholds were calculated as the mean of ipsilateral and contralateral sides. Data are presented as mean ± SEM. Pairwise comparisons were made between male WT vs. male TRPV1 −/− , and male vs. female TRPV1 −/− , n = 7–10. Tests included t‐tests for normally distributed data or Mann–Whitney U ‐test for non‐normal data, ns = not significant, p ≥ 0.05; *** p < 0.001.

Article Snippet: TRPV1 (rabbit, 1:500; Cat. No. ACC‐030, Alomone Labs, Jerusalem, Israel) and NeuN (guinea pig, 1:1000; Cat. No. 266004, Synaptic Systems, Göttingen, Germany) primary antibodies were used to determine TRPV1 expression.

Techniques: Imaging, MANN-WHITNEY

Journal: European Journal of Pain (London, England)

Article Title: Bidirectional Roles of TRPV1 in a Latent Sensitization Model of Myofascial Low Back Pain

doi: 10.1002/ejp.70255

Figure Lengend Snippet: Repeated NGF injections induce manifest mechanical lumbar muscle hypersensitivity. Absolute local PPT values. Y ‐axis shows pressure pain thresholds over lumbar muscles. Arrows indicate NGF/PBS injections (d0, d5). Significant time × treatment interaction (2‐way repeated measures ANOVA) was observed in male WT, male TRPV1 −/− , and female TRPV1 −/− . Post hoc comparisons (NGF vs. PBS) after correction for multiple comparisons are marked, * p < 0.05. Mean ± SEM; n = 5 each, # strong effect, Cohen's d ≥ 1. Time points shown are day 1 after the first injection (d1), day 5 after the first injection (d5), day 1 after the second injection (d1′), day 3 after the second injection (d3′) and day 5 after the second injection (d5′).

Article Snippet: TRPV1 (rabbit, 1:500; Cat. No. ACC‐030, Alomone Labs, Jerusalem, Israel) and NeuN (guinea pig, 1:1000; Cat. No. 266004, Synaptic Systems, Göttingen, Germany) primary antibodies were used to determine TRPV1 expression.

Techniques: Muscles, Injection

Journal: Scientific Reports

Article Title: Potential Orphan Drug Therapy of Intravesical Liposomal Onabotulinumtoxin-A for Ketamine-Induced Cystitis by Mucosal Protection and Anti-inflammation in a Rat Model

doi: 10.1038/s41598-018-24239-9

Figure Lengend Snippet: Alterations of neuroreceptors protein expression in the mucosa layer or smooth muscle layer of the bladder for all groups (n = 8). Western blot analyses with specific antibodies to the TRPV1 receptor and P2X 3 receptor of the rat mucosal layer as well as M 2 -and M 3 - mAChRs and the purinergic P2X 1 receptor of the rat detrusor layer were performed in 3 groups. ( A ) TRPV1 receptor: The TRPV1 antibody produced a clear single band at 95 kDa. ( B ) Purinergic P2X 3 mature receptor: the predominant P2X 3 form (65 kDa). ( C ) M 2 –mAChR of bladder detrusor layer. The M 2 –mAChR antibody produced a clear single band between 50kD and 75kD. ( D ) M 3 –mAChR of bladder detrusor layer. ( E ) Purinergic P2X 1 receptor. Experiments were repeated two times and representative blots are shown. Data of proteins expression (ratios of signal intensities of investigated receptors relative to GAPDH) were calculated with 8 samples in each group. These data of Mean ± SE were standardized and expressed in percentage in which the value of the control group is treated as 100%. Theses values were shown in the bar graph. An asterisk indicates a significant difference between controls and other groups (One-way ANOVA with Dunnett’s test, p < 0.05). The grouping of blots was cropped from the same gel for each protein. The full-length gels and blots are included in the Supplementary Figure .

Article Snippet: Antibodies raised against SNAP25 (1: 1000 dilution; Cell signal), E-cadherin (1:1000 dilution; Cell Signal), nerve growth factor (1:200 dilution; Cell Signal), IL-1β (1:500 dilution; Abcam) IL-6 (1:1000 dilution; Abcam), TNF-α (1:250 dilution, Santa Cruz), NF-κB (1: 2500 dilution; Cell SiZgnal), COX-2 (1:200 dilution; Abcam), TRPV1 receptor (1:1000 dilution; Alomone), purinergic receptor P2X 1 (1:10000 dilution; Alomone), purinergic receptor P2X 3 (1: 1000 dilution; Alomone), M 2 -mAChR (1:1000 dilution; Alomone), M 3 -mAChR (1:1000 dilution; Alomone), and GAPDH (1: 10,000 dilution; Millipore) were used.

Techniques: Expressing, Western Blot, Produced

Journal: Investigative and Clinical Urology

Article Title: Changes in transient receptor potential vanilloid 1 and transient receptor potential vanilloid 4 in patients with lower urinary tract dysfunction

doi: 10.4111/icu.20210418

Figure Lengend Snippet: Comparisons of TRPV1 and TRPV4 expression levels (A) between the DU group and non-DU group and (B) between the urgency group and non-urgency group. TRPV, transient receptor potential vanilloid; DU, detrusor underactivity. * Significant difference between the two groups (p<0.05).

Article Snippet: After two freeze-thaw cycles were performed, the homogenate was centrifuged at 2°C to 8°C for 5 minutes at 5,000× g . The levels of TRPV1 and TRPV4 in the specimen were measured according to the manufacturer’s instructions using a human TRPV1 ELISA kit and a human TRPV4 ELISA kit (Cusabio Biotech Co., Houston, TX, USA).

Techniques: Expressing

Journal: Investigative and Clinical Urology

Article Title: Changes in transient receptor potential vanilloid 1 and transient receptor potential vanilloid 4 in patients with lower urinary tract dysfunction

doi: 10.4111/icu.20210418

Figure Lengend Snippet: Correlation of TRPV1 and TRPV4 with urodynamic parameters

Article Snippet: After two freeze-thaw cycles were performed, the homogenate was centrifuged at 2°C to 8°C for 5 minutes at 5,000× g . The levels of TRPV1 and TRPV4 in the specimen were measured according to the manufacturer’s instructions using a human TRPV1 ELISA kit and a human TRPV4 ELISA kit (Cusabio Biotech Co., Houston, TX, USA).

Techniques:

Journal: Investigative and Clinical Urology

Article Title: Changes in transient receptor potential vanilloid 1 and transient receptor potential vanilloid 4 in patients with lower urinary tract dysfunction

doi: 10.4111/icu.20210418

Figure Lengend Snippet: Immunofluorescence staining of TRPV1 and TRPV4. (A) TRPV1 in the DU group. (B) TRPV1 in the non-DU group. (C) TRPV4 in the DU group. (D) TRPV4 in the non-DU group. (E) TRPV1 in the urgency group. (F) TRPV1 in the non-urgency group. (G) TRPV4 in the urgency group. (H) TRPV4 in the non-urgency group. Scale bars: 200 µm. TRPV, transient receptor potential vanilloid; DU, detrusor underactivity.

Article Snippet: After two freeze-thaw cycles were performed, the homogenate was centrifuged at 2°C to 8°C for 5 minutes at 5,000× g . The levels of TRPV1 and TRPV4 in the specimen were measured according to the manufacturer’s instructions using a human TRPV1 ELISA kit and a human TRPV4 ELISA kit (Cusabio Biotech Co., Houston, TX, USA).

Techniques: Immunofluorescence, Staining

Journal: The Journal of Neuroscience

Article Title: CB1-Dependent Long-Term Depression in Ventral Tegmental Area GABA Neurons: A Novel Target for Marijuana

doi: 10.1523/JNEUROSCI.0190-17.2017

Figure Lengend Snippet: CB1 mediates HFS-LTD. Two candidate receptors that could act as a presynaptic target for eCBs include TRPV1 and CB1. A, The TRPV1 antagonist capsazepine failed to block LTD, suggesting that TRPV1 is not involved in this HFS-LTD (n = 9; p > 0.05 compared with control LTD). B, The CB1 antagonist AM-251 significantly blocked LTD (n = 9, p < 0.05 compared with control LTD).

Article Snippet: A , The TRPV1 antagonist capsazepine failed to block LTD, suggesting that TRPV1 is not involved in this HFS-LTD ( n = 9; p > 0.05 compared with control LTD).

Techniques: Blocking Assay, Control