Properties of subthreshold response and action potential recorded in layer V neurons from cat sensorimotor cortex in vitro

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Abstract
Properties of the action potential and subthreshold response were studied in large layer V neurons in in vitro slices of cat sensorimotor cortex using intracellular recording and stimulation, application of agents that block active conductances, and a single-microelectrode voltage clamp (SEVC). A variety of measured parameters, including action potential duration, afterpotentials, including action potential duration, afterpotentials, input resistance, rheobase and membrane time constant, were similar to the same parameters reported for large neurons from this region of cortex in vivo. Action-potential amplitudes and resting potentials were greater in vitro. Most measured parameters were distributed unimodally, suggesting that these parameters are similar in all layer V neurons irrespective of their axonal termination. The voltage response to subthreshold constant-current pulses exhibited both time and voltage dependence in the great majority of cells. Current pulses in either the hyperpolarizing or subthreshold of depolarizing direction cause the membrane potential to attain an early peak and then decay (sag) to a steady level. On termination of the pulse, the membrane response transiently overshoots resting potential. Plots of current-voltage relations demonstrate inward rectification during polarization on either side of resting potential. Subthreshold inward rectification in the depolarizing direction is abolished by tetrodotoxin (TTX). The ionic currents responsible for subthreshold rectification and sag were examined using the SEVC. Steady inward rectification in the depolarizing direction is caused by a persistent, subthreshold Na current (INaP). Sag observed in response to a depolarizing current pulse is due to activation of a slow outward current, which superimposes on and partially counters the persistent Na current. Both sag in response to hyperpolarizing current pulses and rectification in the hyperpolarizing direction are caused by a slow inward sag current that is activated by hyperpolarizing voltage steps. The sag current is unaltered by TTX; tetraethylammonium, (TEA), Co2+, Ba2+ or 4-aminopyridine. Fast-rising, short-duration action potentials can be elicited by an intracellular current pulse or by orthodromic or antidromic stimulation. Spikes are blocked by TTX. The form of the afterpotential following a directly evoked spike varies among cells with similar resting potentials. Biphasic afterhyperpolarizations (AHP) and fast and slow components were most frequently seen. About 30% of the cells displayed a depolarizing afterpotential (DAP), which was often followed by an AHP. Other cells displayed a purely monophasic AHP. Threshold and accomodative properties of the action potential were studied using linearly rising current ramps. Both voltage and current thresholds remain relatively constant for 0.2- to 1.0-s ramps to the same final current, indicating that accomodation does not occur in this range of ramp durations. The properties of in vitro neocortical neurons examined here are comparable to those measured in vivo by others. For this reason the neocortical slice preparation is a promising system for analysis of the mechanisms underlying neocortical neuron excitability.