Submitted to: Journal of Federation of American Societies for Experimental Biology
Publication Type: Abstract Only
Publication Acceptance Date: December 15, 2009
Publication Date: April 24, 2010
Repository URL: http://www.fasebj.org
Citation: Johnson, W.T., Briske Anderson, M.J., Curtis, S.A., Asher, E.T. 2010. The Influence of Copper (Cu) Deficiency in a Cardiomyocyte Cell Model (HL-1 Cell) of Ischemia/Reperfusion Injury. Journal of Federation of American Societies for Experimental Biology. 24:719.1. Technical Abstract: Mitochondria are important mediators of cell death and this study examines whether mitochondrial dysfunction caused by Cu deprivation promotes cell death in a cell culture model for ischemia/reperfusion injury in cardiomyocytes. HL-1 cells (kindly donated by Dr. William C. Claycomb, LSU Health Sciences Center) were cultured for 3 days in the absence or presence of 80 µM TET, a Cu chelator. TET-treated cells in comparison to untreated cells had lower cytochrome c oxidase activity (32.5±0.4 vs 65.0±4.3 nmol cyt c oxidized·min-1·mg-1) and lower Cu content (0.95±0.06 vs 2.07±0.08 ng Cu/106 cells). After 3 days of TET treatment, ischemia/reperfusion was simulated by replacing medium with nutrient-free buffer and incubating in a hypoxic atmosphere for two hours followed by incubation in complete medium in a normoxic atmosphere for 90 min. Ischemic cells in comparison to control cells had reduced mitochondrial membrane potential (53±1 vs 42±1% depolarized cells) and increased apoptosis (16±1 vs 11±1% apoptotic cells) but TET treatment had no effect. However, following reperfusion, TET- treated compared to untreated cells exhibited lower membrane potential (55±1 vs 46±1% depolarized cells) and increased apoptosis (9.0±0.2 vs 6.9±0.2% apoptotic cells). These findings indicate that Cu status of cardiomyocytes is a determinant of reperfusion injury through a mechanism that likely involves mitochondrial dysfunction.