Veratridine

2-Deoxy-d-glucose (2-DG, Sigma D6134-1G); NMDA (Tocris, 0114); D-AP5 (Tocris, 0106); MK801 (Tocris, 0924); CGP (Tocris, 1493); DIDS (Sigma, D3514); Veratridine (Tocris, 2918); Bumetamide (Sigma, B3023); GlyH-101 (Sigma, 219671); NPPB (Tocris, 0593); Mannitol (Spectrum Chemical, MA165); Ionomycin (Tocris, 1704); SMIFH2 (Sigma, S4826-5MG); CK-666 (Sigma, SML_0006_5MG); Latrunculin A (Lat-A, Sigma, L5163); Thapsigargin (Tocris, 1138); Jasplakinolide (JASP, Tocris, 2792); Bovine Serum Albumin (BSA, Sigma, A4919-5G); DAPI (Biotium, 40043); SAHA (Tocris, 4652); Santacruzamate A (Tocris, 7191); CI994 (Tocris, 2952); FK228 (Tocris, 3515); Tubastatin (Tocris, 6270); Nicotinamide (Tocris, 4106); EX527 (Tocris, 2780); anti-sirtuin 5 inhibitor peptides (Dr.C.A. Olsen, NRE139; NRD167); Suramin (Fisher, AC328540500); C646 (Sigma, SML0002); Cycloheximide (Sigma, C4859); Blebbistatin (Sigma, B0560); Taxol (Tocris, 1097); Nocodazole, (Tocris, 1228); MyoVin-1, (Calbiochem, 475984); Mibefradil, (Tocris, 2198); ω-Conotoxin GVIA, (Tocris, 1085); NBQX (Tocris, 0373); Nifedipine, (Tocris, 1075). Each chemical was dissolved according to the manufacturer’s guidelines, either in water or DMSO, or ethanol.

KB-R7943 (Tocris Bioscience # 1244) was suspended in DMSO and applied to the axonal compartment at a final concentration of 10 μM for 1 h during axotomy (including 15 min pre-incubation before axotomy). Tetrodotoxin citrate (TTX; Tocris Bioscience #1078) was suspended in HBS and applied to the axonal compartment at a final concentration of 1 μM for 1 h during axotomy (beginning 15 min prior to axotomy). Veratridine (Tocris Bioscience #2918) was suspended in DMSO and applied to the axonal compartment at a final concentration of 10 μM for 10 min in the absence of axotomy/injury. Local ABS, which includes low-Ca²⁺, high-Mg²⁺, and TTX (0.5 mM CaCl₂, 10 mM MgCl₂, 1 μM TTX) was applied solely to the axonal compartment for 1 h during axotomy (15 min prior and 45 min after axotomy). Dantrolene (Tocris Bioscience #0507) and (-)-Xestospongin C (Tocris Bioscience #1280), stock concentrations prepared in DMSO, were diluted in HBS or ABS solution and added to axonal compartment at a final concentration of 20 and 1 μM respectively for 1 h during axotomy (with 15 min pre-treatment and 45 min treatment post-axotomy). DMSO or HBS was used as vehicles. Media stored from the axonal compartment prior to treatment was added back to the axonal compartment after treatment and washes with pre-warmed fresh neuron culture media.

Veratridine, dihydrokainic acid (DHK), 2,3-dioxo-6-nitro-1,2,3,4-tetrahydrobenzo[f]quinoxaline-7-sulfonamide disodium salt (NBQX), (S)-α-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (s-AMPA) and citalopram hydrobromide were purchased from Tocris (Bristol, UK). 4-chloro-dl-phenylalanine methyl ester hydrochloride (pCPA) was purchased from Sigma-Aldrich (Tres Cantos, Spain).
For intracerebral microinfusion, Veratridine was dissolved in 12.5% dimethyl sulfoxide in artificial cerebrospinal fluid and pH adjusted to 6.5; DHK and s-AMPA were dissolved in PBS 10 × as previously reported;²⁹ NBQX and citalopram were dissolved in artificial cerebrospinal fluid. The microinfusion volume was 0.5 μl per side in all cases. pCPA was administered intraperitoneally, was daily prepared by dissolving in saline and pH was adjusted to 6.0 with 0.1 m NaOH.^{30 (link)}