aavpro titration kit  (TaKaRa)

Journal: Journal of Neurochemistry

Article Title: Plasticity of Mouse Dorsal Root Ganglion Neurons by Innate Immune Activation Is Influenced by Electrophysiological Activity

doi: 10.1111/jnc.16292

Figure Lengend Snippet: Neurite outgrowth of DRG neurons is diminished by retigabine (RTG) treatment. (A) Example traces of spontaneously firing DRG neurons that become quiescent under RTG treatment. (B) Quantification of action potential (AP) frequency in binned time domains centred around the period of RTG stimulation. (C) Individual traces of unstimulated and RTG‐stimulated APs. Note the deeper AHP curve and slower return to RMP in the RTG trace. (D–G) Representative images of neurites stained with βIII tubulin at the experimental endpoint after treatment with blank media (D, F) or RTG (E, G). (H) Quantification of male and female neurites after treatment with RTG or control (blank media). Scale bar = 50 μm; * p < 0.05. Data is analysed by an unpaired two‐tailed t ‐test. Bar graphs represent mean ± SEM with n = 24 wells from three independent cell culture preparations. Categorical data is represented by parts‐of‐whole transformation.

Article Snippet: For experiments involving the Kv7 channel agonist retigabine (RTG) and the CaMKII inhibitor KN93, a master stock was prepared by reconstituting lyophilised RTG (Tocris, Cat# 6233, 50 mM) or KN93 (Tocris, Cat# 1278, 2.5 mM) in dimethylsulfoxide (DMSO), aliquoting and storing at −20°C.

Techniques: Staining, Control, Two Tailed Test, Cell Culture, Transformation Assay

Journal: Journal of Neurochemistry

Article Title: Plasticity of Mouse Dorsal Root Ganglion Neurons by Innate Immune Activation Is Influenced by Electrophysiological Activity

doi: 10.1111/jnc.16292

Figure Lengend Snippet: Retigabine prevents the growth‐promoting action of inflammatory conditioned media. (A–C) Representative images of neurites from mouse DRG neurons after culture for 48 h with either no treatment, BMDM‐conditioned media treatment, or BMDM‐conditioned media +100 μM RTG treatment. (D) Quantification of mean area of neurite outgrowth. Scale bar = 100 μm, and dashed line represents mean of untreated control; **** p < 0.0001. Data is analysed by one‐way ANOVA. Bar graphs represent mean ± SEM normalised to untreated controls, with n = 6 wells from one cell culture preparation, and the dashed line represents the mean of the untreated control bar.

Article Snippet: For experiments involving the Kv7 channel agonist retigabine (RTG) and the CaMKII inhibitor KN93, a master stock was prepared by reconstituting lyophilised RTG (Tocris, Cat# 6233, 50 mM) or KN93 (Tocris, Cat# 1278, 2.5 mM) in dimethylsulfoxide (DMSO), aliquoting and storing at −20°C.

Techniques: Control, Cell Culture

Journal: Journal of Neurochemistry

Article Title: Plasticity of Mouse Dorsal Root Ganglion Neurons by Innate Immune Activation Is Influenced by Electrophysiological Activity

doi: 10.1111/jnc.16292

Figure Lengend Snippet: Retigabine prevents pain hypersensitivity in the EAE model of neuroinflammatory disease. (A) Withdrawal thresholds to von Frey hair stimulation in mice immunised for EAE and treated with vehicle control (left) or RTG (10 mg/kg) (right). Vehicle‐treated mice exhibit stereotypical reductions in withdrawal thresholds indicative of pain hypersensitivity. Mice that begin treatment with RTG on day 7 post‐immunisation exhibit a complete reversal of these behaviours. * p < 0.05, two‐way ANOVA, Tukey host hoc test. (B) Number of action potentials from DRGNs of EAE‐immunised mice treated with retigabine (RTG, 100 μM) versus vehicle control. Frequency of action potentials (Hz) was recorded from 3 s current injections in 10 pA increments. (C) Resting membrane potential (RMP, mV) from DRGNs of EAE‐immunised mice treated with retigabine (RTG, 100 μM) versus vehicle control. **** p < 0.001, two‐tailed unpaired Student's t ‐test, DRGNs treated with RTG (100 μM) versus vehicle control. (D) Action potential injection threshold (rheobase, pA) from DRGNs of EAE‐immunised mice treated with retigabine (RTG, 100 μM) versus vehicle control. Line graphs represent mean ± SEM, with n = 8 animals from one in vivo experiment. Bar graphs represent mean ± SEM, with n = 24–33 cell recordings from one cell culture preparation. *** p < 0.001, two‐tailed unpaired Student's t ‐test, DRGNs treated with RTG (100 μM) versus vehicle control.

Article Snippet: For experiments involving the Kv7 channel agonist retigabine (RTG) and the CaMKII inhibitor KN93, a master stock was prepared by reconstituting lyophilised RTG (Tocris, Cat# 6233, 50 mM) or KN93 (Tocris, Cat# 1278, 2.5 mM) in dimethylsulfoxide (DMSO), aliquoting and storing at −20°C.

Techniques: Control, Membrane, Two Tailed Test, Injection, In Vivo, Cell Culture