CD38 and Ceramide-Enriched Membrane Platforms in Bovine Coronary - PowerPoint PPT Presentation

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CD38 and Ceramide-Enriched Membrane Platforms in Bovine Coronary Myocytes July 31, 2008 Matt Childrey Department of Pharmacology & Toxicology Medical College of Virginia Virginia Commonwealth University Richmond, Virginia Summer Program

CD38 and Ceramide-Enriched Membrane Platforms in Bovine Coronary Myocytes July 31, 2008 Matt Childrey Department of Pharmacology & Toxicology Medical College of Virginia Virginia Commonwealth University Richmond, Virginia Summer Program for Undergraduate Research 2008
Background • Ceramide-enriched lipid rafts microdomains ceramide sphingomyelin ASM de novo • Oxotremorine CD38 • • 45 kDa
Hypothesis • Lipid raft membrane microdomains are involved in oxotremorine-induced CD38 activation in bovine coronary myocytes. cADPR hydrolysis Calcium CD38 oxotremorine release Lipid raft platform vasoconstriction
Methods •DMT Myograph: tension recording studies of fresh bovine myocardial vasculature •Vasoconstrictor: Oxotremorine (20 uM, 40 uM, 60 uM, 80 uM) •Drugs: Methyl-ß-cyclodextrin (1mM), desipramine (0.1 mM), filipin (1µg/mL), nicotinamide ( 6mM), treated for 15 minutes
Results Oxotremorine-induced Contraction 1000 800 % relative contraction 600 400 200 0 -200 0 uM 20 uM 40 uM 60 uM 80 uM oxotremorine control (n=29) mcd (n=29) desipramine (n=13) filipin (n=16) nicotinamide (n=22) Fig. 1. Effect of LR inhibitors on oxotremorine-induced constriction
Results Methyl-ß-cyclodextrin vs. Control Oxotremorine-induced Contraction (n=29) 700 600 500 % relative contraction 400 300 200 100 0 0 uM 20 uM 40 uM 60 uM 80 uM oxotremorine control mcd
Results Desipramine vs. Control Oxotremorine-induced Contraction 500 400 % relative contraction 300 200 100 0 -100 0 uM 20 uM 40 uM 60 uM 80 uM oxotremorine control (n=29) desipramine (n=13)
Results Filipin vs. Control Oxotremorine-induced Contraction 500 400 % relative contraction 300 200 100 0 0 uM 20 uM 40 uM 60 uM 80 uM oxotremorine control (n=29) filipin (n=16)
Results Nicotinamide vs. Control Oxotremorine-induced Contraction 1000 800 % relative contraction 600 400 200 0 0 uM 20 uM 40 uM 60 uM 80 uM oxotremorine control (n=29) nicotinamide (n=22)
Results
Discussion • CD38 as receptor and enzyme • ASM inhibition • Role of ceramide in CD38 activation? • Raft aggregation and oxotremorine stimulation
Questions and Future Directions • Inhibitor concentrations • Oxotremorine dosage levels • Myograph-specific conditions?
Acknowledgements • Dr.Li and Dr.Zou - special thanks for making this summer possible • VCU Pharmacology & Toxicology

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