Formation and circulation of cerebrospinal fluid (CSF) have been studied in the isolated brain ventricles of anesthetized cats by a new approach and under direct observation. A plastic cannula was introduced into the aqueduct of Sylvius through the vermis cerebelli and the outflow of CSF from the cannula was used as the CSF formation and circulation index. During the 60 min of observation at a physiological CSF pressure not a single drop of CSF escaped out of the end of the cannula. This indicates that CSF net formation and circulation inside the brain ventricles, proposed by classical hypothesis regarding CSF dynamics, should be at least re-evaluated.