Little is known about signaling mechanisms underlying temporal associative learning. Here, we show that mice with a targeted point mutation that prevents autophosphorylation of [alpha]CaMKII ([alpha]CaMKII[superscript T286A]) learn trace eyeblink conditioning normally. This forms a sharp contrast to the severely impaired spatial learning in the water maze and contextual fear conditioning observed in [alpha]CaMKII[superscript T286A] mutants. Importantly, hippocampal lesions impaired trace eyeblink conditioning in [alpha]CaMKII[superscript T286A] mice, suggesting a potential role of hippocampal [alpha]CaMKII-independent mechanisms. These results indicate that hippocampal signaling mechanisms that underlie temporal associative learning as assessed by trace eyeblink conditioning may differ from those of spatial and contextual learning. (Contains 3 figures.)