Increased extracellular ascorbate release reflects glutamate re-uptake during the early stage of reperfusion after forebrain ischemia in rats.
Ascorbate is highly concentrated in neuropils, and its extracellular release is closely related to that of the excitatory neurotransmitters. Thus, the extracellular release of ascorbate and glutamate was measured during the early stage of forebrain ischemia-reperfusion in the rat hippocampus using a microdialysis biosensor system. Male Wistar rats were anesthetized with halothane under mechanical ventilation and normothermia. Two probes of the microdialysis biosensor electrode were inserted in the hippocampus bilaterally. One probe was perfused with phosphate-buffered saline (PBS) and the oxidation signal of dialyzed ascorbate was recorded. A second electropolymerized probe was perfused with PBS containing glutamate oxidase for glutamate measurement. Forebrain ischemia-reperfusion was performed by bilateral carotid artery occlusion with hemorrhagic hypotension (MAP=30 mmHg) for 10 min (Group 10, n=10) or 15 min (Group 15, n=10), followed by reperfusion for 60 min. The release of glutamate increased significantly to 294% (Group 10) and 334% (Group 15) during ischemia, and then decreased rapidly. In Group 15, however, it remained significantly higher after reperfusion than in Group 10. The release of ascorbate increased significantly to 504% (Group 10) and 334% (Group 15) after reperfusion. In Group 10, it was significantly higher for 5-15 min after reperfusion than in Group 15. The marked increase of ascorbate during reperfusion was associated with the rapid decrease in glutamate. The extended time of ischemia significantly inhibited glutamate re-uptake and ascorbate release during reperfusion. These findings suggest the extracellular ascorbate release during reperfusion after global ischemia as a marker of glutamate re-uptake.
Department of Anesthesiology, University of the Ryukyus, Faculty of Medicine, 207 Uehara, Nishihara-cho, Nakagami-gun, 903-0215, Okinawa, Japan. t1935y@med.u-ryuku.ac.jp
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D000818:Animals; D000975:Antioxidants / Q000378:metabolism; D001205:Ascorbic Acid / Q000378:metabolism*; D015374:Biosensing Techniques; D005110:Extracellular Space / Q000378:metabolism; D018698:Glutamic Acid / Q000378:metabolism*; D002546:Ischemic Attack, Transient / Q000378:metabolism*; D008297:Male; D017551:Microdialysis; D016548:Prosencephalon / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D015427:Reperfusion Injury / Q000378:metabolism*; D013997:Time Factors
D000975:Antioxidants; D018698:Glutamic Acid; D001205:Ascorbic Acid