Intracellular recordings revealed that in neocortical explant cultures prepared on the day of birth and examined 3-6 weeks later, neurons mature and establish complex synaptic relationships that lead to spontaneous and triggered synchronous discharge. The spontaneous synchronous activity took several forms, including periodic generation of epileptiform depolarizing waves, prolonged periods of seizure-like discharge, and periodic, intense barrages of IPSPs. Synchronous depolarizations were associated with a marked increase in membrane conductance. Intracellular injection of currents of varying polarity and intensity affected their amplitudes and polarities without influencing the probability of their occurrence, indicating that the discharge reflected the synchronous activities of a neuronal population. This conclusion was confirmed with simultaneous recordings from pairs of neurons. Effects of the GABAa receptor antagonist, bicuculline, and the NMDA receptor antagonist, 2-aminophosphonvalerate (2APV), were used to assess the contributions of impairment of inhibition and enhancement of excitation to the initiation of synchronous discharge. The frequency with which spontaneous depolarizations were generated in normal medium was markedly reduced by 2APV. Moreover, seizure-like activity was induced by removing Mg++ from the medium, a condition that enhances conductance through NMDA receptor-coupled channels. This behavior was also attenuated by 2APV. Perfusion of bicuculline was potently epileptogenic. 2APV cut short the late, voltage-dependent phase of bicuculline-induced paroxysmal depolarizations, indicating a role of NMDA receptors in generating this component of the wave. Epileptiform activities induced by withdrawal of Mg++ were greatly augmented by bicuculline, indicating that blockade of inhibition was not a prerequisite for seizure-like activity. This conclusion is supported by the finding that in many neurons in untreated cultures, paroxysmal generation of trains of IPSPs was the primary manifestation of spontaneous, synchronous population discharge.