Chelation of intracellular calcium reduces cell death after hyperglycemic in vitro ischemia in murine hippocampal slice cultures.

Abstract

The aggravating effect of high glucose levels during cerebral ischemia has been extensively documented in clinical studies and in vivo models of global and focal ischemia. Detailed mechanistic studies of hyperglycemic ischemia have so far been hampered by the lack of in vitro models since glucose during anoxia in vitro is highly protective. We have previously reported glucose toxicity in murine hippocampal organotypic slice cultures exposed to anoxia in an acidotic medium containing high potassium and low calcium. In the present study, we compared the importance of calcium, nitric oxide and free radicals during in vitro ischemia (IVI) and hyperglycemic (40 mM) IVI. Extracellular calcium was a ubiquitous factor for cell death after IVI, but its removal from the medium had no effect on cell death after hyperglycemic IVI. When intracellular calcium was chelated by the 1,2-Bis(2-amino-5-fluorophenoxy)ethane-N,N,N',N'-tetraacetic acid tetrakis(acetoxymethyl) ester (BAPTA-AM) cell death appeared earlier but was mitigated in hyperglycemic IVI, while it was increased in glucose-free IVI. Addition of the nitric oxide synthase (NOS) inhibitor N(omega)-Nitro-L-arginine methyl ester hydrochloride (L-NAME) or the free radical scavengers N-tert-butyl-alpha-phenylnitrone (PBN), deferoxamine and N-acetyl-L-cysteine (NAC) did not affect cell damage in either paradigm. We conclude that the aggravating effect of hyperglycemia during in vitro ischemia is partially mediated by calcium ions released from intracellular stores.

• Authors: Tobias Cronberg, Anna Rytter, Tadeusz Wieloch
• Journal: Brain research (Brain Res) Vol. 1049 Issue 1 Pg. 120-7 (Jul 05 2005) ISSN: 0006-8993 [Print] Netherlands
• PMID: 15935997 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Chelating Agents
• Free Radicals
• 1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester
• Nitric Oxide
• Egtazic Acid
• Glucose
• Calcium

Topics

• Animals
• Calcium (metabolism)
• Cell Death (drug effects)
• Chelating Agents (pharmacology)
• Egtazic Acid (analogs & derivatives, pharmacology)
• Free Radicals (metabolism)
• Glucose (metabolism)
• Hippocampus (drug effects, metabolism, pathology)
• Hyperglycemia (complications, metabolism, pathology)
• In Vitro Techniques
• Intracellular Fluid (drug effects, metabolism)
• Ischemia (etiology, metabolism, pathology)
• Mice
• Mice, Inbred BALB C
• Nitric Oxide (metabolism)