DIFFUSION CHARACTERISTICS AND EXTRACELLULAR VOLUME FRACTION DURING NORMOXIA AND HYPOXIA IN SLICES OF RAT NEOSTRIATUM

1. Diffusion properties of submerged, superfused slices from the rat neostriatum were measured by quantitative analysis of concentration-time profiles of tetramethylammonium (TMA+) introduced by iontophoresis. TMA+ was sensed at an ion-selective microelectrode (ISM) positioned 100-150-mu-m from the source pipette. Slice viability was assessed from the extracellular field potentials evoked by intrastriatal electrical stimulation. 2. Under normoxic conditions the extracellular volume fraction (alpha) was 0.21 (range 0.18-0.24), and the tortuosity (lambda) was 1.54, in slices with good field potentials. In slices with poor field potentials, alpha was 0.09-0.16. Extraction of correct-alpha and lambda in the slice required evaluation of nonspecific uptake, k', which was 1 x 10(-2) s-1. 3. Slices were made hypoxic by superfusing physiological saline equilibrated with 95% N2-5% CO2 for 10-30 min. Synaptic components of field potentials were inhibited after 3-4 min in hypoxic media. In some experiments extracellular K+ concentration ([K+]o) was monitored with ISMs. During hypoxia, [K+]o rose from an average baseline of 5.1 mM to 7-10 mM. After reoxygenation, [K+]o transiently fell below the original level. 4. The average value for alpha during hypoxia was 0.13 (a 38% decrease), which was significantly different from control (P < 0.001) and increased progressively during hypoxic exposure. In contrast, tortuosity and k' were unchanged by this treatment. 5. These data represent the first characterization of the diffusion properties of the rat striatal slice and of changes in extracellular volume fraction during hypoxia in a brain slice preparation. Under normoxic conditions, slices exhibited values for volume fraction and tortuosity that were very similar to those in other preparations. Under hypoxic conditions, alpha was reduced by the same extent as reported in other studies during anoxia in vivo. 6. These results are pertinent to the diffusion of other small molecules in the striatum under conditions where carrier-mediated uptake is largely absent. Two specific applications are the diffusion of striatal dopamine in Parkinson's disease and the accurate assessment of recovery in microdialysis studies.