LGI3/2–ADAM23 interactions cluster Kv1 channels in myelinated axons to regulate refractory period

Isolated dorsal roots were prepared as described previously (Torsney, 2011; Dickie et al., 2017). Following isoflurane-induced anesthesia, control (Adam23LoxP/LoxP; n = 3) and knock-out (PvCre:Adam23LoxP/LoxP; n = 3) mice (∼6–8 wk old) were decapitated and their lumbar (L4/L5) dorsal roots were removed in an ice-cold dissection solution. L4/L5 dorsal roots were cut near their entry zone and their ganglia were removed. The roots were briefly recovered for up to 15 min in 32–34 °C oxygenated NMDG recovery solution and then placed in oxygenated NaCl holding solution for 1 h at room temperature prior to recording. For the electrophysiological compound action potential (CAP) recordings, the isolated roots were transferred to a recording chamber of an upright microscope (Zeiss) and perfused with a constant flow (1–2 ml/min) of oxygenated recording solution. The 95% O2/5% CO2-saturated dissection solution contained 3.0 mM KCl, 1.2 mM NaH2PO4, 26 mM NaHCO3, 15 mM glucose, 251.6 mM sucrose, 7 mM MgCl2, and 0.5 mM CaCl2, pH 7.3–7.4. The recording solution contained 125.8 mM NaCl, 3.0 mM KCl, 1.2 mM NaH2PO4, 26 mM NaHCO3, 15 mM glucose, 1.3 mM MgCl2, and 2.4 mM CaCl2, pH 7.3–7.4. The NMDG recovery solution comprised 93 mM NMDG. 2.5 mM KCl, 1.2 mM NaH2PO4, 30 mM NaHCO3 25 mM glucose, 20 mM HEPES, 5 mM Sodium absorbate, 2 mM Thiourea, 3 mM Sodium pyruvate, 10 mM MgSO4, 0.5 mM CaCl2, pH 7.3–7.4. The holding solution contained 92 mM NaCl, 2.5 mM KCl, 1.2 mM NaH2PO4, 30 mM NaHCO3, 25 mM glucose, 20 mM HEPES, 5 mM Sodium absorbate, 2 mM Thiourea, 3 mM Sodium pyruvate, 2 mM MgSO4, 2 mM CaCl2, pH 7.3–7.4.

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