Journal: Translational Andrology and Urology

Article Title: Evaluation of transurethral GreenLight laser-selective vaporization for refractory overactive bladder in women

doi: 10.21037/tau-24-67

Figure Lengend Snippet: Expressions and comparisons of inflammatory factors and sensory receptors in bladder mucosa tissues before and after treatment. (A) The expressions of TRPV1, P2X3, TNF-α, and IL-6 in patients’ bladder mucosal tissues were stained by immunohistochemical staining before and at 12 weeks after treatment. Representative histological images are shown at 400× magnification. (B) Average optical density value of inflammatory factors and sensory receptors before and after treatment were presented in the bar chart. TNF, tumor necrosis factor; IL-6, interleukin 6; AOD, average optical density.

Article Snippet: Immunohistochemical staining was performed using the avidin-biotin-peroxidase method (ab64212, Abcam, Cambridge, MA, USA) with TRPV1 antibody (CSB-PA822774LA01HU, CUSABIO, Houston, TX, USA), P2X3 antibody (17843-1-AP, Proteintech, Rosemont, IL, USA), tumor necrosis factor alpha (TNF-α) antibody (17590-1-AP, Proteintech, Rosemont, IL, USA), and interleukin 6 (IL-6) antibody (21865-1-AP, Proteintech, Rosemont, IL, USA).

Techniques: Staining, Immunohistochemical staining

Journal: Molecular Pain

Article Title: Knockdown of TRPV2 channels in sensory neurons increases limb use and weight bearing but does not affect spontaneous flinching behavior in a mouse model of bone cancer

doi: 10.1177/1744806918819942

Figure Lengend Snippet: Timeline of experimental protocols for investigating effects of selective inhibition of TRPV2 protein on bone cancer pain (a) or tactile sensory thresholds and thermal nociception in normal mice (b). (a) Two days after insertion of an i.t. catheter, behavioral tests including tests for limb use, weight bearing, and spontaneous flinches were performed as baseline controls. Then, murine sarcoma cells were implanted in the left femurs of mice. Each behavioral test was conducted on days 13, 14, 15, 16, 17, 19, and 21 thereafter. In a separate group of mice, Western blot analysis for expression of TRPV2 protein in dorsal root ganglion neurons was performed at baseline and on days 13, 17, and 19. siRNA against TRPV2 or scrambled siRNA was injected into the i.t. space via the implanted catheter on days 14, 15, and 16 (total dose, 15 µg). (b) Two days after insertion of the i.t. catheter, behavioral tests including tests for tactile sensory thresholds and paw withdrawal latency to noxious heat stimuli were performed at baseline. siRNA against TRPV2 or scrambled siRNA was injected into the i.t. space via the implanted catheter for three consecutive days (total dose, 15 µg). Each behavioral test was performed at baseline and on days 2 and 4 from the last siRNA injection. In another separate group of mice, Western blot analysis for expression of TRPV2 protein in dorsal root ganglion neurons was performed at baseline and on days 2 and 4. siRNA: small interfering RNA; i.t.: intrathecal.

Article Snippet: After washing, the membrane was incubated with an anti-TRPV2 polyclonal antibody (1:2000 dilution; Trans Genic Inc.) and an anti-beta actin rabbit polyclonal antibody (1:5000 dilution; Sigma-Aldrich) overnight at 4°C.

Techniques: Inhibition, Western Blot, Expressing, Injection, Small Interfering RNA

Journal: Molecular Pain

Article Title: Knockdown of TRPV2 channels in sensory neurons increases limb use and weight bearing but does not affect spontaneous flinching behavior in a mouse model of bone cancer

doi: 10.1177/1744806918819942

Figure Lengend Snippet: TRPV1 and TRPV2 expression in dorsal root ganglion (DRG) neurons innervating bone marrow of the femur. We injected the retrograde neurotracer FG into bone marrow of the left femurs of normal mice (5 µl of 2% FG solution). Forty-eight hours after the FG injection, lumbar DRGs were removed for TRPV1 and TRPV2 immunohistochemistry. FG-labeled DRG (a and c), TRPV1-ir (b), and TRPV2-ir DRG neurons (d) are also shown. The sections in (a) and (b) and those in (c) and (d) are the same. The section of TRPV1-ir (b) is different from that of TRPV2-ir (d). Arrows indicate FG-labeled DRG neurons (a and c), and arrowheads indicate double-labeled DRG neurons (b and d). Size distribution of TRPV1-ir FG-labeled DRG neurons (e). Size distribution of TRPV2-ir FG-labeled DRG neurons (f). Profiles of the DRG neurons were divided into small (<30 µm), medium (30–40 µm), and large (>40 µm) cells by measuring the diameters. FG-labeled DRG neurons, TRPV1-ir FG-labeled DRG neurons or TRPV2-ir FG-labeled DRG neurons were counted in each section. Then, the total numbers of neurons in the 24 sections derived from eight mice (L2–L4 DRGs) were estimated. TRPV1: transient receptor potential vanilloid subfamily 1; TRPV2: transient receptor potential vanilloid subfamily 2; ir: immunoreactive; FG: Fluoro-Gold.

Article Snippet: After washing, the membrane was incubated with an anti-TRPV2 polyclonal antibody (1:2000 dilution; Trans Genic Inc.) and an anti-beta actin rabbit polyclonal antibody (1:5000 dilution; Sigma-Aldrich) overnight at 4°C.

Techniques: Expressing, Injection, Immunohistochemistry, Labeling, Derivative Assay

Journal: Molecular Pain

Article Title: Knockdown of TRPV2 channels in sensory neurons increases limb use and weight bearing but does not affect spontaneous flinching behavior in a mouse model of bone cancer

doi: 10.1177/1744806918819942

Figure Lengend Snippet: Effects of intrathecal administration of siRNA against TRPV2 (TRPV2-siRNA) and scr-siRNA on bone cancer pain-related behaviors. Murine sarcoma cells were injected into mouse femurs. From day 14, mice were treated with TRPV2-siRNA or scr-siRNA for three consecutive days (arrows). Scores of limb use during spontaneous ambulation (a) and weight bearing during spontaneous standing (b) in the TRPV2-siRNA group after siRNA administration were significantly higher than those in the scr-siRNA group. In contrast, there was no significant difference in the number of spontaneous flinches between the two groups (c). n = 8, in each group. *P < 0.05 versus scr-siRNA. BL: baseline; TRPV2: transient receptor potential vanilloid subfamily 2; siRNA: small interfering RNA; scr: scrambled control.

Article Snippet: After washing, the membrane was incubated with an anti-TRPV2 polyclonal antibody (1:2000 dilution; Trans Genic Inc.) and an anti-beta actin rabbit polyclonal antibody (1:5000 dilution; Sigma-Aldrich) overnight at 4°C.

Techniques: Injection, Small Interfering RNA

Journal: Molecular Pain

Article Title: Knockdown of TRPV2 channels in sensory neurons increases limb use and weight bearing but does not affect spontaneous flinching behavior in a mouse model of bone cancer

doi: 10.1177/1744806918819942

Figure Lengend Snippet: Changes in TRPV2 protein expression in lumbar dorsal root ganglion neurons ipsilateral to the femur injected with sarcoma cells. Small interfering RNA (siRNA) against TRPV2 was injected into the intrathecal space on days 14, 15, and 16. (a) Representative results of Western blot analysis of TRPV2 (upper) and beta-actin (lower). (b) Quantitative results of TRPV2 protein expression. TRPV2 protein expression was normalized to beta-actin levels. TRPV2 protein expression did not change until day 13. TRPV2 protein expression was significantly downregulated by siRNA against TRPV2. Data are expressed as means ± standard deviations ( n = 4). *P < 0.05 versus BL. #P < 0.05 versus day 13. BL: baseline; TRPV2: transient receptor potential vanilloid subfamily 2.

Article Snippet: After washing, the membrane was incubated with an anti-TRPV2 polyclonal antibody (1:2000 dilution; Trans Genic Inc.) and an anti-beta actin rabbit polyclonal antibody (1:5000 dilution; Sigma-Aldrich) overnight at 4°C.

Techniques: Expressing, Injection, Small Interfering RNA, Western Blot

Journal: Molecular Pain

Article Title: Knockdown of TRPV2 channels in sensory neurons increases limb use and weight bearing but does not affect spontaneous flinching behavior in a mouse model of bone cancer

doi: 10.1177/1744806918819942

Figure Lengend Snippet: Effects of intrathecal administration of siRNA against TRPV2 (TRPV2-siRNA) and scr-siRNA on thermal sensitivity and tactile sensory thresholds of normal mice without cancer. TRPV2-siRNA or scr-siRNA was administered for three consecutive days (arrows). On the second, third, and fourth days after completion of siRNA administration, we investigated the thermal sensitivity and tactile sensory thresholds in normal mice. TRPV2-siRNA did not affect the mechanical withdrawal threshold (a) or withdrawal latency to thermal stimulation (b) in the paws of normal mice, as compared with those treated with scr-siRNA. n = 8, in each group. BL: baseline; TRPV2: transient receptor potential vanilloid subfamily 2; siRNA: small interfering RNA; scr: scrambled control.

Article Snippet: After washing, the membrane was incubated with an anti-TRPV2 polyclonal antibody (1:2000 dilution; Trans Genic Inc.) and an anti-beta actin rabbit polyclonal antibody (1:5000 dilution; Sigma-Aldrich) overnight at 4°C.

Techniques: Small Interfering RNA

Journal: Molecular Pain

Article Title: Knockdown of TRPV2 channels in sensory neurons increases limb use and weight bearing but does not affect spontaneous flinching behavior in a mouse model of bone cancer

doi: 10.1177/1744806918819942

Figure Lengend Snippet: Changes in TRPV2 protein expression in lumbar dorsal root ganglion neurons in normal mice. siRNA against TRPV2 was injected into the intrathecal space via the implanted catheter for three days. (a) Representative results of Western blot analysis of TRPV2 (upper) and beta-actin (lower). (b) Quantitative results of TRPV2 protein expression. TRPV2 protein expression was normalized to beta-actin levels. TRPV2 protein expression was decreased at two and four days after intrathecal injection of siRNA against TRPV2. Data are expressed as means ± standard deviations (n = 4). *P < 0.05 versus BL. BL: baseline; TRPV2: transient receptor potential vanilloid subfamily 2; siRNA: small interfering RNA.

Article Snippet: After washing, the membrane was incubated with an anti-TRPV2 polyclonal antibody (1:2000 dilution; Trans Genic Inc.) and an anti-beta actin rabbit polyclonal antibody (1:5000 dilution; Sigma-Aldrich) overnight at 4°C.

Techniques: Expressing, Injection, Western Blot, Small Interfering RNA