L-citrulline, L-ornithine, and agmatine are the metabolites of L-arginine by nitric oxide synthase (NOS), arginase, and arginine decarboxylase (ADC), respectively. In contrast to the NOS and arginase pathways, the role of the ADC-agmatine pathway in learning and memory has only been paid attention lately. Recent evidence suggests a potential involvement of agmatine in learning and memory processing. The present study further addressed this issue by comparing the levels of agmatine, as well as L-arginine, L-citrulline, and L-ornithine, in the hippocampus, parahippocampal region, prefrontal cortex, vestibular nucleus, and cerebellum in rats that were trained in the delayed nonmatch to position task in the T-maze with their yoked controls. There were significantly increased agmatine levels in the prefrontal, entorhinal, and perirhinal cortices and increased L-citrulline concentrations in the dentate gyrus (DG) and prefrontal cortex in the T-maze training group relative to the control one. L-arginine and L-ornithine levels were not significantly different between groups in the brain regions examined. These results demonstrate T-maze training-induced region-specific increases in L-citrulline and agmatine. Significant positive correlations between prefrontal and perirhinal agmatine levels and animals' performance in the T-maze further suggest the direct involvement of agmatine in learning and memory processing.