Cell death in an ischemic infarct rim model.
Using an in vitro model that simulates the microenvironment in the ischemic infarct rim, we have examined the temporal profile and possible mechanisms of cell death in the neuropil (an astrocyte-rich area or ARA) of organotypic hippocampal slice cultures. Two-photon confocal microscopy, propidium iodide, and GFAP-GFP transgenic mice were used to confirm cell death in astrocytes. An 'ischemic solution' (IS) induced major cell death throughout the hippocampus over 24 h, with the earliest injury starting in ARA. Our studies using IS or ion replacements in IS revealed that cell death in ARA was modest when K(+) was increased or pH lowered. High K(+) is most effective in reducing cell death when HCO(3)(-) is normal or high. When Cl(-) or HCO(3)(-) was reduced, cell injury was worsened. 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS) protected cells from IS-induced death in a dose-dependent manner (1-4000 micromol/L). We conclude that (i) various areas of the hippocampal formation respond differently to ionic replacements; (ii) K(+) interacts with other ions to protect cells in ARA; and (iii) DIDS has a substantial protective effect in ARA by blocking DIDS-sensitive membrane exchangers or by interfering with intracellular signaling pathways.
Yao
Hang
H
Department of Pediatrics (Section of Respiratory Medicine), University of California, San Diego, La Jolla, California, USA.
Sun
Xiaolu
X
Gu
Xiang
X
Wang
Juan
J
Haddad
Gabriel G
GG
D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid / Q000494:pharmacology; D000818:Animals; D016923:Cell Death / Q000187:drug effects / Q000502:physiology; D002544:Cerebral Infarction / Q000235:genetics / Q000473:pathology / Q000517:prevention & control; D020925:Hypoxia-Ischemia, Brain / Q000235:genetics / Q000473:pathology* / Q000517:prevention & control; D051379:Mice; D008822:Mice, Transgenic; D009924:Organ Culture Techniques; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics
D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid