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• W2313631279 abstract "BACKGROUND: Calcium (Ca2+) is a cofactor of multiple cellular processes. The mechanisms that lead to elevated cytosolic Ca2+ concentration are unclear. OBJECTIVE: To illuminate how bloody cerebrospinal fluid (bCSF) from patients with intraventricular hemorrhage causes cell death of cultured human astrocytes. METHODS: Cultured astrocytes were incubated with bCSF. In control experiments, native CSF was used. Cytosolic Ca2+ concentration was measured by fura-2 fluorescence. Apoptosis and necrosis were evaluated by staining with Hoechst-3342 and propidium iodide. RESULTS: Incubation of astrocytes with bCSF provoked a steep Ca2+ concentration peak that was followed by a slow Ca2+ rise during the observation period of 50 minutes. Necrosis, but not apoptosis, was induced. Blockade of ATP-sensitive P2 receptors with suramin inhibited the bCSF-induced initial Ca2+ peak and necrosis. Blockade of P1 receptors with 8-phenyltheophylline or of N-methyl-D-aspartate receptors with D(−)-2-amino-5-phosphopentanoic acid had no significant effect. Preincubation with xestospongin D, a blocker of inositol 1,4,5-trisphosphate receptors, prevented the initial Ca2+ rise and reduced the rate of necrosis. Preemptying of the endoplasmic reticulum with thapsigargin protected astrocytes from the bCSF-induced Ca2+ peak. Inhibition of mitochondrial permeability transition pores opening with cyclosporin A reduced the rate of astrocytic necrosis significantly, although it did not influence the initial Ca2+ peak. CONCLUSION: bCSF elicits a steep, transient Ca2+ rise when administered to human astrocytes by activation of ATP-sensitive P2 receptors and subsequent inositol 1,4,5-trisphosphate-dependent Ca2+ release from endoplasmic reticulum. This massive Ca2+ overload leads to subsequent mitochondrial permeability transition pores opening and necrosis of the cells. ABBREVIATIONS: bCSF, bloody cerebrospinal fluid ER, endoplasmic reticulum HEPES, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid HOE, Hoechst 33342 nCSF, native cerebrospinal fluid NMDA,N-methyl-D-aspartate PI, propidium iodide SAH, subarachnoid hemorrhage SERCA, endoplasmic reticulum calcium ATPase TBI, traumatic brain injury XeD, xestospongin D" @default.
• W2313631279 created "2016-06-24" @default.
• W2313631279 creator A5021967295 @default.
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• W2313631279 date "2013-03-01" @default.
• W2313631279 modified "2023-10-18" @default.
• W2313631279 title "The Mechanisms of Energy Crisis in Human Astrocytes After Subarachnoid Hemorrhage" @default.
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• W2313631279 doi "https://doi.org/10.1227/neu.0b013e31827d0de7" @default.
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