Variations in the excitability of individual cortical neurones during the invasion of spreading depression (SD) have been monitored by observing the alterations of spontaneous and L-glutamate-induced firing. Invasion of many neurones during SD is marked by a brief burst of firing which occurs concurrently with the onset of the negative slow extracellular potential. Other neurones do not fire, although the microelectrode records a negative slow wave. Depression of glutamate-induced and spontaneous firing follows and may last for several minutes. The initial loss of excitability of those neurones that discharge during SD invasion may be due to excessive depolarization. This phase is rapidly succeeded by a period of depressed excitability, during which the neurones can be invaded by an antidromic spike or excited by increased amounts of L-glutamate. These findings indicate that SD propagation initially involves the release of an excitant substance, possibly glutamic acid. The continuing effects of SD are due to the reduction in cell excitability. As many neurones are depressed without undergoing an initial excitation, it appears that a depressant substance is also involved. This may be gamma-aminobutyric acid.