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: British Journal of Pharmacology

Article Title: Targeting nociceptive transient receptor potential channels to treat chronic pain: current state of the field

doi: 10.1111/bph.14044

Figure Lengend Snippet: Schematic diagrams of TRP channels, showing two of the four subunits in the tetrameric structures. All panels follow the same colour scheme from N‐ to C‐terminus: N‐terminal cytoplasmic domain, turquoise; linker, yellow; S1–S4, lilac; S5–S6, blue; TRP‐box helix, red; C‐terminus, green; and flexible N‐ or C‐terminal extensions, grey. (A) TRPV1, TRPV3 and TRPV4 proteins contain six ankyrin repeats in their N‐terminus. The C‐terminus interacts with the ankyrin repeats, connecting the cytoplasmic regions. A chemical agonist (e.g. capsaicin or resiniferatoxin for TRPV1) binding site was identified at the interface of the S1–S4 and S5–S6 regions. Of note, TRPV4 contains a proline‐rich region in its flexible N‐terminal extension. (B) TRPA1's unique structural features include a C‐terminal coiled‐coil region and 17 ankyrin repeats in the N‐terminus; repeats 1–11 were not modelled in the cryoEM structure and are likely to connect to repeats 12–17 through a flexible link. An IP6 molecule bridges the N‐terminal ankyrin repeats to the C‐terminal coiled‐coil. A chemical antagonist, A967079, binds within the S5–S6 pore region (yellow circle). Cysteines sensing reactive electrophiles are represented as black circles in the membrane‐proximal N‐terminal linker region that surrounds the TRP‐box helix (red). (C) No structure is yet available for TRPM8, but its transmembrane domain is homologous to that of TRPV1 and TRPA1. The N‐terminal region contains ‘melastatin homology regions’ (MHRs), and its C‐terminus contains a predicted coiled coil.

Article Snippet: Not unexpectedly, TRPV1 channels are activated by painful venoms from a wide range of species including jellyfish (Cuypers et al., 2006 ), spiders (Siemens et al., 2006 ) and centipedes (Yang et al., 2015 ).

Techniques: Binding Assay, Membrane

Journal: British Journal of Pharmacology

Article Title: Targeting nociceptive transient receptor potential channels to treat chronic pain: current state of the field

doi: 10.1111/bph.14044

Figure Lengend Snippet: Chemical structures of TRPV1 channel modulators. Capsaicin and resiniferatoxin are agonists, while AMG‐517 and AZD1386 are antagonists.

Article Snippet: Not unexpectedly, TRPV1 channels are activated by painful venoms from a wide range of species including jellyfish (Cuypers et al., 2006 ), spiders (Siemens et al., 2006 ) and centipedes (Yang et al., 2015 ).

Techniques: