Low dose of D-cycloserine (DCS), a partial agonist of glycine binding site on N-methyl-D-aspartate (NMDA) receptors, can facilitate extracellular signal-regulated kinase1/2 (ERK1/2) activity in the amygdala and modulate emotional behavior. However, the relationship between ERK1/2 activation, individual anxiety levels, and DCS is unknown. Therefore, based on open arm time in the elevated plus-maze, male Wistar rats were divided into subgroups with either low (LOA) or high open arm (HOA) time. Open arm time is usually accepted as a critical index of unconditioned anxiety-like/avoidance behavior. On the following day, DCS (30 mg/kg, i.p.) was administered 30 min before the second elevated plus-maze test. On day 8 and 9, the rats were subjected to a 2-day session of the forced swim test, receiving the DCS treatment again 30 min before the 2nd day. On the 16th day, 30 min after the administration of DCS, the rats were sacrificed in order to detect the phosphorylation of ERK1/2 (p-ERK1/2) in the amygdala by Western blots. The results showed that: (1) DCS decreased the open arm time in HOA but not LOA rats. (2) DCS suppressed the immobility in the day-2 trial of the forced swim test and increased the p-ERK1/2 level in the amygdala in LOA but not HOA rats. This is the first instance data has been found indicating different sensitivities of p-ERK1/2 and behavioral responses to the treatment of DCS between HOA and LOA rats. The results suggest that the activity of NMDA receptor-mediated ERK1/2 signaling is mediated by individual behavioral differences which are related to the antidepressant-like activity of DCS. This study provides first insight into the pathophysiological role of ERK signaling with regard to individual differences in emotional behavior.