1 The ability of bath-applied L-glutamate to enhance subsequent depolarizations produced by bath-applied L-aspartate on lobster muscle was further investigated by means of intracellular recording techniques. 2. Increasing the conditioning glutamate concentration or exposure time produced a greater enhancement of aspartate responses. Enhancement was also dependent on the time interval between glutamate and aspartate doses and was not prevented by overnight storage of preparations in vitro. 3. The dose-depolarization curve for enhanced aspartate responses (measured at a fixed time following a given dose of glutamate) was displaced to the left along the abscissa scale relative to control, with no detectable change in limiting log-log slope. 4. Conditioning doses of kainate or domoate (but not quisqualate, aspartate, or KCl) also enhanced aspartate responses; however, their conditioning effect was little affected by increasing the concentration, exposure time, or time interval before applying aspartate. The rate of onset and decline of the enhanced aspartate response always resembled that of the previous conditioning agonist. 5. D and L-Aspartate were approximately equieffective depolarizing agents whereas D-glutamate was approximately 1/40 as potent as L-glutamate. After a conditioning dose of D or L-glutamate, responses to D or L-aspartate were enhanced. 6. In a Na+-free (Li+) medium, both the glutamate depolarization and the conditioning effect towards aspartate were largely abolished. With kainate however, Na+ was not apparently important either for evoking the kainate response or for producing the conditioning effect. 7. Bath-applied glutamate greatly enhanced and prolonged the time course of the iontophoretic aspartate potential with only a small effect on the glutamate potential; however, these effects were not maintained after washout of glutamate. In contrast, bath-application of aspartate depressed the aspartate potential while enhancing the glutamate potential. Some sites that were insensitive to iontophoretically-applied aspartate became clearly responsive to this agent during a bath-application of glutamate. 8. It is proposed that during conditioning with bath-applied glutamate, kainate or domoate, some agonist is trapped by extrajunctional sites and is subsequently displaced by bath-applied aspartate to produce the long-term enhancement effect.