Developmental changes in Na⁺ conductances in rat neocortical neurons: Appearance of slowly inactivating component

1. Na⁺ conductances have been characterized in rat neocortical neurons from the sensorimotor area. Neurons were obtained by acute dissociation from animals at developmental stages from embryonic day 16 (E16) to postnatal day 50 (P50) to quantify any developmental changes in the kinetic properties of the Na⁺ conductance. 2. Neurons were divided into two classes, based on morphology, to determine whether there are any cell-type specific differences in Na⁺ conductances that contribute to the different action potential morphologies seen in current-clamp recordings in vitro. 3. Upon isolation, neurons were voltage clamped using the whole-cell variation of the patch-clamp technology. Both cell types, pyramidal and nonpyramidal, demonstrate large increases in Na⁺ current density during this developmental period (E16-P50). Normalized conductances were near 10 pS/μm² in neurons from embryonic animals, and increased 6- to 10-fold during the first 2 wk postnatal. The final conductance reached in pyramidal neurons was higher than in nonpyramidal neurons. 4. We found no differences between the two cell types, pyramidal and nonpyramidal, in the boltage dependence of activation, inactivation kinetics, voltage dependence of steady-state inactivation, and recovery from inactivation. 5. The time course of Na⁺ current in immature neurons were fit with classical Hodgkin-Huxley kinetics. However, in more mature neurons the kinetics of inactivation became more complicated such that two decay components were required to obtain good fit. The slowly decaying component had a time course 5 to 10 times slower than the fast component. 6. Several procedures were used to reduce the magnitude of Na⁺ conductance in mature neurons to ensure graded, voltage-dependent inward currents. These included reduced extracellular [Na⁺], submaximal tetrodotoxin concentrations, and reduced holding potential. Under each of these conditions we were able to verify the observation that Na⁺ current inactivation occurs with two exponentials. 7. Single-channel Na⁺ currents were obtained from cell-attached patches. The membrane density of active Na⁺ channels increases with development, and ensemble averages from mature neurons demonstrated two inactivation processes. The slow inactivation process was accounted for by long-latency single-channel openings of the same amplitude as the short-latency openings. 8. We conclude that there are no kinetic differences in the Na⁺ channels between cell types. Differences in action potentials are then not explained by differences in Na⁺ current kinetics, but might be partially explained by the different densities. The developmental increase in Na⁺ current density, which is related to an increase in the number of channels per unit membrane area, underlies the increased rate of rise of action potentials in neurons during maturation. The increase in a slowly inactivating component with development affects near-threshold functions such as the persistent inward current that contributes to repetitive firing.