Electrophysiological properties and synaptic responses in the deep layers of the human epileptogenic neocortex in vitro

Abstract

1. Neocortical slices of the first and second temporal gyrus and frontal lobe, removed in human epileptic patients for the relief of intractable seizures, were maintained in vitro at 35 +/- 1 degrees C. Electrophysiological properties of neurons in the deep layers (1,800–2,600 micron below the pial surface) were studied with conventional intracellular recording and stimulation techniques. Synaptic responses were evoked by extracellular focal stimuli. Intracellular injections of some cells with the fluorescent dye Lucifer yellow revealed large spiny pyramidal neurons. 2. Values of input resistance, resting membrane potential (Vm), and action-potential amplitude were similar for neurons in different cortical regions. These parameters were also similar when neurons were grouped in accordance to the degree of electrographic epileptiform activity displayed by the cortical tissue in situ. 3. Inward rectification occurred when neurons were depolarized by 5–15 mV positive to the resting Vm. This rectification was abolished by extracellular application of tetrodotoxin (TTX, 1 microM), but was still observed in the presence of the Ca2+-channel blocker Cd2+ (2 mM). Pulses of hyperpolarizing current elicited a slowly developing inward rectification, called anomalous rectification, which was insensitive to TTX, but blocked by extracellular application of Cs+ (1-2 mM). 4. Intracellular injection of depolarizing square pulses of current (0.1-4 s) evoked repetitive firing. In most cells the firing rate decreased smoothly for tens of milliseconds (i.e., it adapted) before reaching a steady level. Plots of the relation between frequency of the repetitive firing and injected current (f-I curve) displayed two linear segments for the early intervals as well as for the adapted and/or the steady firing. The slope of the initial, steeper linear segment of the f-I curve computed during the early intervals and during the adapted firing was 163 +/- 51 and 56 +/- 27 (SD) Hz/nA, respectively. 5. A long-lasting (up to 8 s) afterhyperpolarization (AHP) followed the repetitive firing induced by square pulses of depolarizing current. Its amplitude was directly proportional to the amount of current injected, it was sensitive to changes in the Vm, and it had an equilibrium potential 10–40 mV negative to the resting Vm. This value plus the fact that the AHP could be recorded with KCl-filled microelectrodes suggested that it was caused by an increase in conductance to K+ ions. Bath application of the Ca2+ channel blockers Cd2+ (2 mM) or Mn2+ (2 mM) decreased and eventually blocked the AHP.(ABSTRACT TRUNCATED AT 400 WORDS)