rabbit polyclonal antibody against human trpv1  (Santa Cruz Biotechnology)

Journal: Annals of the Rheumatic Diseases

Article Title: Increased function of pronociceptive TRPV1 at the level of the joint in a rat model of osteoarthritis pain

doi: 10.1136/annrheumdis-2013-203413

Figure Lengend Snippet: Transient receptor potential vanilloid 1 (TRPV1) and CD68 immunoreactivities in human synovium. TRPV1 (A) and CD68 (B) immunoreactivities in synovium from a patient with osteoarthritis (OA). Immunoreactivities both for TRPV1 (green) and for CD68 (red) colocalise to mononuclear cells within the synovial lining and sublining regions (arrows). Occasional CD68 immunoreactive cells do not display TRPV1 immunoreactivity (arrowheads). L: surface of synovial lining. Double fluorescence immunohistochemistry. Scale bar=50 μm. (C) Box and whisker plots showing increased abundance of TRPV1-immunoreactive cells in OA compared with non-arthritic postmortem (PM) control synovium. **p=0.003. (D) Western blot analysis of TRPV1 in membrane-enriched fractions of synovial from five patients with OA. TRPV1 immunoreactivity was detected at approximately 100 kDa in lanes 1, 3, 4 and 5.

Article Snippet: Immunohistochemistry with an affinity-purified rabbit polyclonal antibody directed against human TRPV1 (SC-20813, Santa Cruz Biotechnology Inc.) was conducted on synovial sections from patients with OA (n=9; three men/six women), RA (n=8; two men/six women) and PM controls (n=7; six men/one woman).

Techniques: Fluorescence, Immunohistochemistry, Whisker Assay, Control, Western Blot, Membrane

Journal: Annals of the Rheumatic Diseases

Article Title: Increased function of pronociceptive TRPV1 at the level of the joint in a rat model of osteoarthritis pain

doi: 10.1136/annrheumdis-2013-203413

Figure Lengend Snippet: Endogenous transient receptor potential vanilloid 1 (TRPV1) activation at the level of the joint contributes to pain. (A) Intra-articular administration of JNJ-17203212 significantly attenuated weight-bearing asymmetry compared with intra-articular vehicle (3% Tween 80, 0.5% EtOH in saline) in rats 14 days post-mono-iodoacetate (MIA) injection (n=11/9 rats) and to a similar degree as systemic (intra-peritoneal (i.p)) JNJ-17203212. Vehicle effects following intra-articular and i.p administration were not significantly different; for clarity, intra-articular vehicle is presented. Data are presented as mean±SEM and analysis used a two-way analysis of variance (ANOVA) with a Bonferroni's post hoc test (weight-bearing) or Mann–Whitney test (allodynia). *p<0.05 between i.art JNJ-17203212 and i.art vehicle. $ p<0.05 between i.p JNJ-17203212 and i.art vehicle. (B) Levels of the endogenous TRPV1 ligand 12-hydroxy-eicosatetraenoic acid were significantly greater in joint tissue from MIA-treated rats compared with saline-treated rats (p<0.05, Mann–Whitney; n=8 rats per group). (C) Stimulus–response curves of joint afferents to mechanical von Frey monofilament stimulation (0.16–15g) applied to the centre of the receptive field in MIA-treated and saline-treated rats at day 14 post-injection. There was a significant leftward shift in the stimulus–response curve in MIA-treated rats compared with saline-treated rats (**p<0.01, n=30/37 MIA/saline, respectively). Data are displayed at mean±SEM. Analysis used a two-way ANOVA with Bonferroni's post hoc test. (D) Close intra-arterial administration of the TRPV1 agonist capsaicin (5–10 μM) increased (sensitised) mechanically evoked responses of joint afferents in saline-treated rats. A significant increase in the frequency of firing was observed with 5 μM capsaicin in 40% (8/20) of fibres in saline-treated rats (***p<0.001, n=8). Data are displayed at mean±SEM. Analysis used a two-way ANOVA with Bonferroni's post hoc test. (E) Close intra-arterial administration of capsaicin (5 and 10 μM) significantly reduced mechanical thresholds of 45% (9/20) of joint afferents in saline-treated rats (n=9). Horizontal bars represent median values and analysis used a Mann–Whitney test, *p<0.05.

Article Snippet: Immunohistochemistry with an affinity-purified rabbit polyclonal antibody directed against human TRPV1 (SC-20813, Santa Cruz Biotechnology Inc.) was conducted on synovial sections from patients with OA (n=9; three men/six women), RA (n=8; two men/six women) and PM controls (n=7; six men/one woman).

Techniques: Activation Assay, Saline, Injection, MANN-WHITNEY

Journal: Annals of the Rheumatic Diseases

Article Title: Increased function of pronociceptive TRPV1 at the level of the joint in a rat model of osteoarthritis pain

doi: 10.1136/annrheumdis-2013-203413

Figure Lengend Snippet: Transient receptor potential vanilloid 1 (TRPV1) immunoreactivity in the spinal cord. TRPV1 immunofluorescence detected in superficial dorsal horn (10× magnification) in rat lumbar (L3–L5) spinal cord at day 28 post-intra-articular injection of saline (A) or mono-iodoacetate (MIA) (B). Minimum and maximum brightness values were altered (32.01 min and 90.14 max) using Image J so as to highlight the area of TRPV1 positive staining. (C) Quantification of TRPV1 immunofluorescence in superficial dorsal horn of spinal cord taken from rats at 14 or 28 days following intra-articular injection of MIA and at day 28 following intra-articular injection of saline. Data are expressed as mean and SEM (n=5 per group). Analysis used a two-way analysis of variance (ANOVA) with a Bonferroni's post hoc test. *p<0.05 and ***p<0.001 compared with saline treatment and ###p<0.001 between day 14 and day 28 MIA treatment.

Article Snippet: Immunohistochemistry with an affinity-purified rabbit polyclonal antibody directed against human TRPV1 (SC-20813, Santa Cruz Biotechnology Inc.) was conducted on synovial sections from patients with OA (n=9; three men/six women), RA (n=8; two men/six women) and PM controls (n=7; six men/one woman).

Techniques: Immunofluorescence, Injection, Saline, Staining