Glycine-activated currents in 1- to 11-day-old rat medullary neurons were studied using patch clamp techniques. Glycine produced neither repeatable whole-cell current responses nor single-channel activity in the cell-attached mode until cells were in culture for a week or more. However, Cl- channels were present at the early stages because glycine-activated channels were seen in excised, inside-out patches. Furthermore, for cells less than a week in culture, 10 patches which did not exhibit glycine-activated Cl- channels in the cell-attached mode did upon excision. Consequently, the activation properties of these Cl- channels undergo a developmental change in that some cellular factor(s) presumably prevents the Cl- channels from opening in the intact cell during the initial stages in culture.