Investigation of Phospholipase C-β1b Splice Variants on Function in H9c2 Cells Chelsea Parker Dr. Theresa Filtz Oregon State University HHMI Program Summer.

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Congestive Heart Failure

Presentation transcript:

Investigation of Phospholipase C-β1b Splice Variants on Function in H9c2 Cells Chelsea Parker Dr. Theresa Filtz Oregon State University HHMI Program Summer 2009

Importance Hypertrophy Congestive Heart Failure Causes: obesity, birth defects, high blood pressure Widespread: over 79 million people affected Treatments: Digitalis, Beta blockers, diuretics

Background Phospholipase C-β’s are stress activated enzymes present in many tissues and organs Only the tail structure of PLC-β1b differs from other phospholipase C-β subtypes

Molecular Pathway PLC-β1b in heart cells Signaling

Previous Research -Control PLC  1a PLC  1b +Control Cell area ** No virus PLC  1a PLC  1b

Project Description Study H9c2 cells Induce hypertrophy Assay changes in native PLC- β1 levels Transfect H9c2 cells with both PLC-β1 variants Assay for hypertrophy Create mutants of PLC-β1b Over-express the PLC-β1b mutants in H9c2 cells Assay for hypertrophy

Hypothesis The tail section (extreme C-terminus) of PLC-β1b is responsible for PLC-β1b -induced heart cell enlargement Prediction: Over-expression of PLC-β1b tail segments will lead to decreased heart cell enlargement in H9c2 cells

Results H9c2 cell maintenance techniques Identification of PLCB-1 in H9c2 cells Hypertrophy Transformation of GFP, PLC-β1a, and PLC-β3 Transfection and expression of GFP in H9c2 cells 7/14/2009

Continued Research Optimize transfection efficiency Generate a PLC-β1b construct with PCDNA as the vector Create PLC-β1b mutants Transfect H9c2 cells with PLC-β DNA and PLC-β1b mutant DNA

Special Thanks To Dr. Theresa Filtz Dr. Satin Salehi Dr. Kevin Ahern Howard Hughes Medical Institute Ray, Frances, and Dale Cripps Scholarship Fund