Poly l ornithine hbr

Sensory neurons of trigeminal ganglia (TGs) were obtained from 8 to 12 week-old Trpm3^+/+ and Trpm3^−/− mice, as described before [32 (link),37 (link)]. Briefly, mice were euthanized by CO₂, TGs were isolated and digested with collagenase (2 mg/ml) and dispase (2,5 mg/ml) (both from Invitrogen/Thermo Fisher Scientific, Waltham, MA, USA). Suspension of sensory neurons was seeded on laminin (100 μg/ml) and poly-L-ornithine HBr (500 μg/ml) (both from Sigma Aldrich) coated glass bottom culture dishes (MatTek, Ashland, MA, USA) and cultured in Neurobasal medium supplemented with 2% B-27 supplement, 2 mM L-glutamine, 100 μg/ml penicillin/streptomycin, 2 ng/ml glial cell line-derived neurotrophic factor (GNDF) (all from Invitrogen/Thermo Fisher Scientific) and 10 ng/ml NT-4 (PeproTech, London, UK) at 37 °C in 5% CO₂ containing humidified atmosphere. Neurons were used for experiments within 24 to 36 hrs following isolation.