Late calcium EDTA rescues hippocampal CA1 neurons from global ischemia-induced death.
Transient global ischemia induces a delayed rise in intracellular Zn2+, which may be mediated via glutamate receptor 2 (GluR2)-lacking AMPA receptors (AMPARs), and selective, delayed death of hippocampal CA1 neurons. The molecular mechanisms underlying Zn2+ toxicity in vivo are not well delineated. Here we show the striking finding that intraventricular injection of the high-affinity Zn2+ chelator calcium EDTA (CaEDTA) at 30 min before ischemia (early CaEDTA) or at 48-60 hr (late CaEDTA), but not 3-6 hr, after ischemia, afforded robust protection of CA1 neurons in approximately 50% (late CaEDTA) to 75% (early CaEDTA) of animals. We also show that Zn2+ acts via temporally distinct mechanisms to promote neuronal death. Early CaEDTA attenuated ischemia-induced GluR2 mRNA and protein downregulation (and, by inference, formation of Zn2+-permeable AMPARs), the delayed rise in Zn2+, and neuronal death. These findings suggest that Zn2+ acts at step(s) upstream from GluR2 gene downregulation and implicate Zn2+ in transcriptional regulation and/or GluR2 mRNA stability. Early CaEDTA also blocked mitochondrial release of cytochrome c and Smac/DIABLO (second mitochondria-derived activator of caspases/direct inhibitor of apoptosis protein-binding protein with low pI), caspase-3 activity (but not procaspase-3 cleavage), p75NTR induction, and DNA fragmentation. These findings indicate that CaEDTA preserves the functional integrity of the mitochondrial outer membrane and arrests the caspase death cascade. Late injection of CaEDTA at a time when GluR2 is downregulated and caspase is activated inhibited the delayed rise in Zn2+, p75NTR induction, DNA fragmentation, and cell death. The finding of neuroprotection by late CaEDTA administration has striking implications for intervention in the delayed neuronal death associated with global ischemia.
Calderone
Agata
A
Department of Neuroscience, Albert Einstein College of Medicine, Bronx, New York 10461, USA.
Jover
Teresa
T
Mashiko
Toshihiro
T
Noh
Kyung-min
KM
Tanaka
Hidenobu
H
Bennett
Michael V L
MV
Zukin
R Suzanne
RS
D000818:Animals; D051017:Apoptosis Regulatory Proteins; D002545:Brain Ischemia / Q000378:metabolism / Q000473:pathology*; D002352:Carrier Proteins / Q000378:metabolism; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects / Q000502:physiology; D002614:Chelating Agents / Q000494:pharmacology*; D045304:Cytochromes c / Q000378:metabolism; D053938:DNA Fragmentation / Q000187:drug effects; D004492:Edetic Acid / Q000494:pharmacology*; D005849:Gerbillinae; D006624:Hippocampus / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D008297:Male; D008928:Mitochondria / Q000378:metabolism; D024101:Mitochondrial Proteins / Q000378:metabolism; D009474:Neurons / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D020800:Receptor, Nerve Growth Factor; D018091:Receptors, AMPA / Q000378:metabolism / Q000502:physiology; D017475:Receptors, Nerve Growth Factor / Q000378:metabolism; D013997:Time Factors; D015032:Zinc / Q000378:metabolism / Q000502:physiology*
D051017:Apoptosis Regulatory Proteins; D002352:Carrier Proteins; D002614:Chelating Agents; C489013:DIABLO protein, rat; D024101:Mitochondrial Proteins; D020800:Receptor, Nerve Growth Factor; D018091:Receptors, AMPA; D017475:Receptors, Nerve Growth Factor; D045304:Cytochromes c; D004492:Edetic Acid; C505410:Casp3 protein, rat; D053148:Caspase 3; D020169:Caspases; D015032:Zinc; C104722:glutamate receptor ionotropic, AMPA 2