, 2010). Measuring the Na+ current directly at the mammalian node of Ranvier, however, will require further improvements in optical and electrical recording methods. Although the present study was done in neocortical L5 axons, this functional role for the first node might be found in
other myelinated axons of the central nervous system. For instance, in neurons of the inferior Neratinib olive, it was recently noted that the AP number in a burst critically depended on the length of the remaining axon (Mathy et al., 2009). Similar to the present findings, only neurons with axons longer than ∼100 μm, spared by the slice-cutting procedure, generated multiple spikes in a single burst. Since olivary axons have an AIS length of ∼40 μm, at which point it becomes myelinated (de Zeeuw et al., 1990), it is conceivable that the intrinsic complex spike burst in olivary axons requires the activation of nodal Na+ channels, in analogy with the present observations. In summary, the present data demonstrate that Na+ channels in the first node of Ranvier of mammalian neocortical axons have a direct contribution to the information processing capacity by increasing the spike output gain near threshold and facilitating high-frequency APs. These findings add to a growing literature revealing that axons are highly enriched, with voltage-gated mechanisms enabling complex integrative operations and a dynamic regulation
of AP initiation and patterns (Debanne et al., 2011). Pomalidomide nmr All experiments were carried out according to guidelines approved
by the Animal Ethics Committee of the Australian National University. Adult male Wistar rats (22–42 days old) were deeply anaesthetized with 3% isoflurane and decapitated. Parasagittal cortical brain slices were cut with until an angle of 15° at 300 μm (Vibratome 7000s, Campden Instruments Ltd., Loughborough, UK) in a high-Mg2+/low-Ca2+ ice-cold artificial cerebrospinal fluid (ACSF) consisting of 125 mM NaCl, 25 mM NaHCO3, 3 mM KCl, 1.25 mM NaH2PO4, 25 mM glucose, 1 mM CaCl2, and 6 mM MgCl2 (pH 7.4, oxygenated with 95% O2/5% CO2). Individual slices were transferred to the stage of a Zeiss Axioskop (Carl Zeiss, NSW, Australia), and L5 pyramidal neurons were visualized using Dodt optics (Luigs & Neumann Elektrotechnik, Ratingen, Germany). Slices were perfused with oxygen-saturated (95% O2, 5% CO2) ACSF consisting of 125 mM NaCl, 25 mM NaHCO3, 3 mM KCl, 1.25 mM NaH2PO4, 25 mM glucose, 2 mM CaCl2, and 1 mM MgCl2. Whole-cell current-clamp recordings were made using either Dagan BVC-700A (Dagan Corporation, Minneapolis, MN), Axoclamp 2A, or Multiclamp 700A amplifiers (Molecular Devices, Inc., Sunnyvale, CA) in bridge mode. Voltage was analog low-pass filtered at 10 kHz (Bessel) and digitally sampled at 50 or 100 kHz using an ADA converter (ITC-18, Heka Elektronik, Lambrecht, Germany), and data were acquired with Axograph X software (AxoGraph Scientific, Sydney, NSW, Australia).