DRG neurons were excised from 6-week-old mice (WT or TRPV1−/−) and enzymatically dissociated in HBSS containing collagenase (2 mg/ml) and dispase (4 mg/ml; Invitrogen) for 45 min at 37°C (20). DRGs were rinsed twice in HBSS and once in culture medium consisting of Dulbecco’s minimum essential medium supplemented with 10% heat-inactivated FBS, streptomycin (100 μg/ml), penicillin (100 U/ml), and nerve growth factor (100 ng/ml; all from Invitrogen). Individual neurons were dispersed by trituration through a fire-polished glass Pasteur pipette in 4 ml of media. Neurons were cultured on glass coverslips previously treated with HBSS + 25% poly-L-ornithine and laminin (both from Sigma), overnight at 37°C with 5% CO2 in 96% humidity.

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