1. We have determined the ascorbic acid (ascorbate) and glutathione (GSH) content of cortical and hippocampal slices form rat brain after prolonged (6 h) incubation and have correlated these levels with the histological quality of the slices. Ascorbate and GSH levels in control and sliced tissue were determined by high performance liquid chromatography (HPLC) with electrochemical detection. Cell morphology of incubated slices was compared with that of intact tissue in cresyl violet stained tissue sections. 2. Roughly 70% of tissue ascorbate and GSH was lost from slices during incubation in vitro. Normal in vivo levels of ascorbate (2-3 μmol g-1 tissue wet weight) could be maintained by including 200-400 μM ascorbate (typical extracellular concentration) in the incubation media. By contrast, the loss of GSH could not be prevented by incubation with GSH. 3. The morphology of cells in hippocampal slices incubated under conditions that maintained ascorbate content and compartmentalization were similar to those of intact tissue. Ascorbate protected pyramidal cells in CA1 and CA3 regions of the hippocampus from the degeneration that was seen in slices incubated in ascorbate-free media. 4. These data suggest that loss of endogenous antioxidants may be a major factor in neuronal loss in vitro and support the notion that ascorbate is an endogenous neuroprotective agent.
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