The visual system of Hermissenda has been studied extensively as a site of cellular plasticity produced by classical conditioning. A one-trial conditioning procedure consisting of light paired with the application of serotonin (5-HT) to the exposed, but otherwise intact, nervous system produces suppression of phototactic behavior tested 24 h after conditioning. Short- and long-term enhancement (STE and LTE) of excitability in identified type B photoreceptors is a cellular correlate of one-trial conditioning. LTE can be expressed in the absence of STE suggesting that STE and LTE may be parallel processes. To examine the development of enhancement, we studied its time-dependent alterations after one-trial conditioning. Intracellular recordings from identified type B photoreceptors of independent groups collected at different times after conditioning revealed that enhanced excitability follows a biphasic pattern in its development. The analysis of spikes elicited by 2 and 30 s extrinsic current pulses at different levels of depolarization showed that enhancement reached a peak 3 h after conditioning. From its peak, excitability decreased toward baseline control levels 5-6 h after conditioning followed by an increase to a stable plateau at 16 to 24 h postconditioning. Excitability changes measured in cells from unpaired control groups showed maximal changes 1 h posttreatment that rapidly decremented within 2 h. The conditioned stimulus (CS) elicited significantly more spikes 24 h postconditioning for the conditioned group as compared with the unpaired control group. The analysis of the time-dependent development of enhancement may reveal the processes underlying different stages of memory for this associative experience.