Investigation of Phospholipase C-β1b Splice Variants on Function in H9c2 Cells

Chelsea Parker Dr. Theresa Filtz

Oregon State UniversityHHMI 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

http://odyb.net/physical-health/9-types-of-diseases-caused-by-obesity/

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

http://atlasgeneticsoncology.org/Genes/PLCB1ID41742ch20p12.html

Molecular Pathway

PLC-β1b in heart cells Signaling

http://journals.prous.com/journals/dof/19992407/html/df240759/images/Yerxa2.gif

Previous Research

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PLC

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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