Journal:

Article Title: Inhibition of m3 muscarinic acetylcholine receptors by local anaesthetics

doi: 10.1038/sj.bjp.0704040

Figure Lengend Snippet: (A) m3 muscarinic signalling induced by MCh (4.2×10−7 M) is not affected by relevant concentrations of QX314. Only concentrations of 10−3 M (78±8.7% of control response) and 10−2 M (55±5.1% of control response) inhibit partially. (B) Schematic illustration of the chimeras employed. (C) Concentration-response curve for effect of extracellularly applied QX314 on chimera 1. IC50 is 1.8±0.6 μM. Mean control response was 4.9±0.4 μA. (D – E) Effects of various concentrations of extracellularly administered QX314 on chimera 3 (D) and chimera 5 (E). Modest inhibition was observed in both cases (* indicates significant differences vs control). Maximal inhibition is obtained using 10−3 M QX314 on chimera 3 (64.2±7.5% of control response (1.9±0.2 μA)) and using 10−4 M QX314 on chimera 5 (58.2±8.5% of control response (1.5±0.2 μA)). (F) Extracellularly applied QX314 (10−3 M) does not inhibit chimera 2, 4 or 6. Mean response sizes are 102.1±8.5, 96.2±15 and 104.8±13.5% of control response for chimera 2, 4 and 6, respectively.

Article Snippet: Sincere thanks to Astra Pharmaceuticals, LP (Westborough, MA, U.S.A.) for supplying QX314.

Techniques: Control, Concentration Assay, Inhibition

Journal:

Article Title: Inhibition of m3 muscarinic acetylcholine receptors by local anaesthetics

doi: 10.1038/sj.bjp.0704040

Figure Lengend Snippet: Concentration-dependent inhibition of MCh (4.2×10−7 M)-induced ICl(Ca) in oocytes expressing m3 muscarinic receptors by (A) intracellularly injected QX314 (IC50 445±226 μM), (B) extracellularly administered benzocaine (IC50 258±182 μM) and (C) intracellularly applied lidocaine (IC50 341±33 μM).

Article Snippet: Sincere thanks to Astra Pharmaceuticals, LP (Westborough, MA, U.S.A.) for supplying QX314.

Techniques: Concentration Assay, Inhibition, Expressing, Injection

Journal:

Article Title: Inhibition of m3 muscarinic acetylcholine receptors by local anaesthetics

doi: 10.1038/sj.bjp.0704040

Figure Lengend Snippet: (A) Responses to MCh (4.2×10−7 M) after selective G-protein α-subunit knockdown using oligonucleotides directed against Gαo, Gαq, Gα11 or Gα14, as compared with control responses. Data were collected 48 h after antisense injection. Knockdown of Gαq (49.3±5.7% of control) or Gα11 (65.4±6.8% of control) significantly affected MCh-induced responses (P<0.05), whereas anti-Gαo (110.7±13.6% of control) or anti-Gα14 (112.9±12.9% of control) injected cells showed responses similar to those observed in control cells (P<0.05). Therefore, m3 signalling is mediated primarily by Gq and G11. (B) Mean±s.d. of ICl(Ca), elicited by MCh (10−7 M) in oocytes expressing m3 muscarinic receptors, injected with 50 nl of 150 mM KCl (control, 3.4±0.16 μA, black bar); QX314 (445×10−6 M) reduced m3 responses to 44±3% of the control response (1.51±0.11 μA, white bar) 150 mM KCl 48 h after injection of either anti-Gαq (1.43±0.18 μA) or anti-Gα11 (1.35±0.24 μA, grey bars); or QX314 (445×10−6 M) 48 h after injection of either anti-Gαq (1.55±0.13 μA) or anti-Gα11 (0.5±0.11 μA, white bars). Lack of effect of the local anaesthetic after Gq knockdown indicates that inhibition requires this G proteins subunit. (C) Schematic summary of hypothesized sites of action on m3 muscarinic signalling for the local anaesthetics studied.

Article Snippet: Sincere thanks to Astra Pharmaceuticals, LP (Westborough, MA, U.S.A.) for supplying QX314.

Techniques: Knockdown, Control, Injection, Expressing, Inhibition

Journal: The Journal of Clinical Investigation

Article Title: Mechanisms and treatments of neuropathic itch in a mouse model of lymphoma

doi: 10.1172/JCI160807

Figure Lengend Snippet: ( A ) Response latency in hot-plate tests at 50°C, 53°C, and 56°C in vehicle-treated ( n = 10) and RTX-treated ( n = 9) CTCL mice at day 20. F (2, 51) = 98.25, P < 0.0001. ( B ) CTCL-induced itch in the early phase (day 20) in mice before (baseline [BL])and after the treatment with vehicle ( n = 6) or RTX ( n = 6). F (1, 10) = 42.93, P < 0.0001. ( C ) CTCL-induced itch in mice in the late phase (day 60) before and after the treatment with vehicle ( n = 6) or RTX ( n = 6). F (1, 10) = 52.77, P < 0.0001. ( D ) CTCL-induced itch in the early phase (day 20) in mice before and 1 hour and 3 hours after the treatment with intratumoral injection of vehicle, flagellin (Fla) (1 μg, 30 μL), QX-314 (6 mM, 30 μL), QX-314 (6 mM, 30 μL) plus flagellin (1 μg), or QX-314 (6 mM, 30 μL) plus capsaicin (Cap) (10 μg). n = 6/group. F (4, 75) = 4.49, P = 0.0026. ( E ) CTCL-induced itch in the late phase (day 60) in mice before and 1 hour and 3 hours after intratumoral injection of vehicle, flagellin (1 μg, 30 μL), QX-314 (6 mM, 30 μL), QX-314 (6 mM, 30 μL) plus flagellin (1 μg), or QX-314 (6 mM, 30 μL) plus capsaicin (10 μg). n = 6/group. F (4, 22) = 4.20, P = 0.0112. Data are expressed as the mean ± SEM. Two-way ANOVA with Bonferroni’s post hoc test, # P < 0.05, ## P < 0.01, ### P < 0.001, and *** P < 0.001.

Article Snippet: Gabapentin (catalog 60142-96-3), capsaicin (catalog 404-86-4), QX-314 (catalog 21306-56-9), and RTX (catalog R-6712) were purchased from LC Laboratories.

Techniques: Injection

Journal: Journal of Pain Research

Article Title: Comparison of the transport of QX-314 through TRPA1, TRPM8, and TRPV1 channels

doi: 10.2147/JPR.S41614

Figure Lengend Snippet: Chemical structure of FAM-conjugated QX-314 (molecular mass, 692 Da). Abbreviations: FAM, 5(6)-carboxyfluorescein; QX-314, N-ethyl-lidocaine.

Article Snippet: QX-314 (Enzo Life Sciences Inc) was dissolved in physiological saline.

Techniques:

Journal: Journal of Pain Research

Article Title: Comparison of the transport of QX-314 through TRPA1, TRPM8, and TRPV1 channels

doi: 10.2147/JPR.S41614

Figure Lengend Snippet: Effect on thermal sensitivity in TRPA1 channels opening. Notes: injections (10 μL in each group) of only 5% AITC (AITC group), only 2% QX-314 (QX-314 group), a mixture of AITC and QX-314 (AITC/QX-314 group), or the vehicle (vehicle group) were made into rat right plantar hind paws. The experimental groups were not significantly different from the vehicle group. Abbreviations: AITC, allyl isothiocyanate; QX-314, N-ethyl-lidocaine; TRPAI, transient receptor potential ankyrin I.

Article Snippet: QX-314 (Enzo Life Sciences Inc) was dissolved in physiological saline.

Techniques:

Journal: Journal of Pain Research

Article Title: Comparison of the transport of QX-314 through TRPA1, TRPM8, and TRPV1 channels

doi: 10.2147/JPR.S41614

Figure Lengend Snippet: Effect on thermal sensitivity in TRPM8 channels opening. Notes: Injections (10 μL in each group) of only 5% menthol (menthol group), only 2% QX-314 (QX-314 group), a mixture of menthol and QX-314 (menthol/QX-314 group), or the vehicle (vehicle group), were made into rat right plantar hind paws. The menthol/QX-314 group showed more analgesia than the vehicle group at 10 to 60 minutes ( P < 0.01). The menthol group showed more analgesia than the vehicle group at 10 to 60 minutes ( P < 0.01). No significant difference was seen between the menthol group and the menthol/QX-314 group. The results indicate the independent analgesic effect of menthol. Abbreviations: QX-314, N-ethyl-lidocaine; TRPM8, transient receptor potential melastatin-8.

Article Snippet: QX-314 (Enzo Life Sciences Inc) was dissolved in physiological saline.

Techniques:

Journal: Journal of Pain Research

Article Title: Comparison of the transport of QX-314 through TRPA1, TRPM8, and TRPV1 channels

doi: 10.2147/JPR.S41614

Figure Lengend Snippet: Effect on thermal sensitivity in TRPV1 channels opening. Notes: Injections (10 μL in each group) of only 0.1% capsaicin (capsaicin group), only 2% QX-314 (QX-314 group), a mixture of capsaicin and QX-314 (capsaicin/QX-314 group), or the vehicle (vehicle group), were made into rat right plantar hind paws. The capsaicin/QX-314 group showed longer withdrawal latency than the QX-314 group or the vehicle group at 60 to 300 minutes ( P < 0.01, and P < 0.001, respectively). The capsaicin/QX-314 group showed analgesia compared with the capsaicin group at 60 to 180 minutes ( P < 0.05, and P < 0.01, respectively). The capsaicin group showed analgesia compared with vehicle group at 240 to 300 minutes ( P < 0.01). The QX-314 group did not differ significantly from the vehicle group. Abbreviations: QX-314, N-ethyl-lidocaine; TRPV1, transient receptor potential ankyrin.

Article Snippet: QX-314 (Enzo Life Sciences Inc) was dissolved in physiological saline.

Techniques:

Journal: Journal of Pain Research

Article Title: Comparison of the transport of QX-314 through TRPA1, TRPM8, and TRPV1 channels

doi: 10.2147/JPR.S41614

Figure Lengend Snippet: Inhibition of the antinociceptive effect of capsaicin/QX-314 by capsazepine. Notes: Capsazepine (50 μg/mL; 10 μL) was injected 30 minutes prior to the capsaicin/QX-314 (10 μL) injection (capsazepine/capsaicin/QX-314 group) into rat right plantar hind paws. The capsaicin/QX-314 group showed a longer withdrawal latency than the capsazepine/capsaicin/QX-314 group, the QX-314 group, or the vehicle group at 60 to 300 minutes ( P < 0.01, P < 0.001). The capsazepine/capsaicin/QX-314 group did not differ significantly from the vehicle group. Abbreviation: QX-314, N-ethyl-lidocaine.

Article Snippet: QX-314 (Enzo Life Sciences Inc) was dissolved in physiological saline.

Techniques: Inhibition, Injection

Journal: Journal of Pain Research

Article Title: Comparison of the transport of QX-314 through TRPA1, TRPM8, and TRPV1 channels

doi: 10.2147/JPR.S41614

Figure Lengend Snippet: Effect on uptake of the fluorescence tracer (FAM). The fluorescence in small neurons (arrows) was conspicuous in DRGs injected with capsaicin and QX-314-FAM ( A ). Fluorescence was not detected in DRGs injected with only QX-314-FAM ( B ), AITC/QX-314-FAM ( C ), and menthol/QX-314-FAM ( D ). Abbreviations: AIT, allyl isothiocyanate; DRG, dorsal root ganglia; FAM, 5(6)-carboxyfluorescein; QX-314, N-ethyl-lidocaine; TRPA1, transient receptor potential ankyrin 1; TRPM8, transient receptor potential melastatin-8; TRPV1, transient receptor protein vanilloid 1.

Article Snippet: QX-314 (Enzo Life Sciences Inc) was dissolved in physiological saline.

Techniques: Fluorescence, Injection