Cell volume regulation has been studied in neuronal and glial cultures but little is known about volume regulation in brain tissue with an intact extracellular space. We investigated volume regulation in hippocampal slices maintained in an interface chamber and exposed to hypo-osmotic medium. Relative changes in intracellular and extracellular volume were measured respectively as changes in light transmittance and extracellular resistance. Slices exposed to hypo-osmotic medium (200-240 mOsm/L) showed a decrease in light transmittance, which occasionally was preceded by a brief transient increase. However, hypo-osmotic exposure was always accompanied by a monotonic increase in extracellular resistance. Peak changes in light transmittance and extracellular resistance occurred at 15-20 min following exposure to hypo-osmotic medium. Optical evidence of volume regulation (RVD) was observed in six of 12 slices and occurred over the next 60-90 min. We hypothesized that the relatively low incidence of RVD was related to depletion of taurine, an osmolyte known to play an important role in volume regulation, during preparation of the slices. Indeed, taurine levels in freshly prepared slices were <50% of those reported in intact hippocampus. Incubation of slices in 1 mM taurine restored taurine to levels observed in situ and increased both the likelihood and magnitude of RVD in hypo-osmotic medium. Inhibition of taurine flux with 100 microM 5-nitro-2-(3 phenylpropylamino) benzoic acid blocked both RVD and the transient undershoot of volume commonly associated with return of swollen slices to iso-osmotic medium. Taurine treatment had no effect on levels of several other amino acids but preserved slice potassium content. The results indicate a critical role for cellular taurine during hypo-osmotic volume regulation in hippocampal slices. Inconsistencies between optical measurements of cellular volume changes and electrical measurements of extracellular space are likely to result from the complex nature of light transmittance in the interface slice preparation.