Journal: Pain

Article Title: Inhibition of dual leucine zipper kinase prevents chemotherapy-induced peripheral neuropathy and cognitive impairments

doi: 10.1097/j.pain.0000000000002256

Figure Lengend Snippet: The DLK inhibitor IACS′8287 prevents cisplatin induced peripheral neuropathy. (A) Treatment schedule of IACS′8287 and cisplatin. IACS′8287 (30 mg/kg) or vehicle via oral gavage was given 1 day before start of each cisplatin cycle and 1 hour before each cisplatin or PBS i.p. injection for 2 treatment cycles. (B) Representative Western blot and quantification for p-c-Jun, c-Jun, and GAPDH in DRG from mice received one cycles of IACS′8287 and cisplatin treatment (n = 8, two-way ANOVA reveal significant main effect on interaction: F (1, 28) = 4.601, P = 0.0408, and significant IACS′8287 treatment effect: F (1, 28) = 58.55, P < 0.0001; * P < 0.05, ** P < 0.01, **** P < 0.0001). Sensitivity to mechanical stimulation of the hind paws was monitored over time using the von Frey test. Coadministration of IACS′8287 prevented cisplatin-induced mechanical allodynia in (C) male mice (n = 8, statistical analysis was conducted using Log[withdrawal threshold (g)], two-way repeated-measures ANOVA reveal significant main effect on interaction: F (9, 84) = 105, P < 0.0001, significant treatment effect: F (3, 28) = 98.73, P < 0.0001, and significant time effect: F (3, 84) = 438.8, P < 0.0001; Tukey post hoc analysis: *** P < 0.001, **** P < 0.0001 as compared to Veh+PBS; ^^^^ P < 0.0001 as compared to IACS′8287+Cisplatin) and (D) female mice (statistical analysis was conducted using Log[withdrawal threshold (g)], two-way repeated-measures ANOVA reveal significant main effect on interaction: F (6, 27) = 15.78, P < 0.0001, significant treatment effect: F (2, 9) = 13.72, P = 0.0018, and significant time effect: F (3, 27) = 89.31, P < 0.0001; Tukey post hoc analysis: **** P < 0.0001 as compared to Veh+PBS; ^^^^ P < 0.0001 as compared to IACS′8287+Cisplatin). Paw biopsies were obtained from the hind paws of mice that received 2 cycles of cisplatin with or without the DLK inhibitor IACS′8287. Tissues were stained with antibodies against PGP9.5 (red) and collagen (green). (E) Representative images from each treatment group, scale bar = 20 μm, magnification ×40. (F) Quantification of IENF density expressed as the number of nerve fibers crossing the basement membrane/length of the basement membrane (mm) (n = 8, two-way ANOVA reveal significant main effect on interaction: F (1, 28) = 12.68, P = 0.0013; Tukey post hoc analysis: * P < 0.05, ** P < 0.01). (E) Lumbar DRG neurons were isolated after completion of behavioral testing for mitochondrial bioenergetic analysis using the Seahorse XFe24 Flux Analyzer. Changes induced by cisplatin and IACS′8287 in (G) basal OCR, MRC, and SRC (n = 6, two-way ANOVA reveal significant main effect on interaction: F (1, 20) = 18.88, P = 0.0003 for basal OCR; F (1, 20) = 22.57, P = 0.0001 for MRC; F (1, 20) = 17.70, P = 0.0004 for SRC; Tukey post hoc analysis: * P < 0.05). Results are expressed as mean ± SEM. ANOVA, analysis of variance; DLK, dual leucine zipper kinase; DRG, dorsal root ganglion; IENF, intraepidermal nerve fibers; MRC, maximal respiratory capacity.

Article Snippet: Primary antibodies against DLK (75-355, 1:500, NeuroMab, Davis, CA), phospho-c-Jun (Ser63) (9261, 1:250, Cell Signaling Technology, Danvers, MA), c-Jun (9156, 1:250, Cell Signaling Technology, Danvers, MA), or glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (MAB374, 1:5000, Millipore, Burlington, MA) were used, followed by LI-COR secondary antibody (1:5000).

Techniques: Injection, Western Blot, Staining, Membrane, Isolation

Journal: eLife

Article Title: Neuronal hyperexcitability is a DLK-dependent trigger of herpes simplex virus reactivation that can be induced by IL-1

doi: 10.7554/eLife.58037

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-DLK/MAP3K12 (Rabbit polyclonal) , Thermo Fisher , PA5-32173 RRID: AB_2549646 , WB (1:500).

Techniques: Recombinant, Plasmid Preparation, Blocking Assay, Sequencing, shRNA, Control, SYBR Green Assay, Staining