Two monkeys were trained in a delayed sequential motor task in which the time interval between events and the delay duration were either fixed or variable. Single-unit neuronal activity was recorded in the pre-supplementary motor area (pre-SMA). During the delay, we observed a gradual increase in activity (build-up pattern) in the fixed but not in the variable condition. In the former but not in the latter, the monkey had the opportunity to estimate time duration. Consequently, the build-up pattern observed in the pre-SMA might represent the neuronal substrate of a time accumulator system proposed by previous authors on the basis of functional imaging data. Such a system could play a critical role in the working memory of temporal information.