|Budget Amount *help
¥3,000,000 (Direct Cost : ¥3,000,000)
Fiscal Year 1998 : ¥1,500,000 (Direct Cost : ¥1,500,000)
Fiscal Year 1997 : ¥1,500,000 (Direct Cost : ¥1,500,000)
Hippocampal slices including entorhinal cortex were cut from 8-9 day-old Wistar rat brain and cultivated for 2 weeks in the medium consisting of 50 % MEM, 25 % HBSS and 25 % horse serum. Glucose deprivation or exposure to excitatory amino acid were employed as a model of ischemic brain cell death. In the former experiment, glucose-deficient HBSS containing 10mM 2-deoxyglucose was used for 1 h as glucose-deprivation. Dead neurons were quantified with 5 μM propidium iodide (PI) 48 h after the restoration period in normal medium. In the latter experiment, N-methyl-D-aspartate (NMDA) was included in the normal medium for short period (13 - 100 μM, 15 min). Dead neurons were evaluated 24 and 48 h after the exposure. Glucose deprivation for 1 h induced CA1 area specific neuronal cell death. α,β-methylene ATP (1 mM) included in the medium both during and after the glucose-deprivation period inhibited this neuronal cell death, however, this agent did not affect cell survival by itself. Short-term exposure to NMDA induced concentration-dependent and area-dependent neuronal cell death, i.e. CA1>CA3>dentate gyrus. ATP and purinergic agents, applied both during and after the NMDA-exposure period, did not affect the NMDA-induced neuronal cell death in any hippocamlap area. These results of this research projects indicate that purinergic agents might be utilized as novel neuroprotective therapeutics in hypoglycemic ischemia.