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Investigation of Phospholipase C-β1b Splice Variants on Function in H9c2 Cells
Chelsea Parker Dr. Theresa Filtz
Oregon State UniversityHHMI Program Summer 2009
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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/
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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
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Molecular Pathway
PLC-β1b in heart cells Signaling
http://journals.prous.com/journals/dof/19992407/html/df240759/images/Yerxa2.gif
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Previous Research
-Control
PLC1a PLC1b
+Control Cell
are
a
**
0.0
1.0
2.0
No vir
us
PLC
1a
PLC
1b
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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
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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
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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
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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
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Special Thanks To
Dr. Theresa Filtz
Dr. Satin Salehi
Dr. Kevin Ahern
Howard Hughes Medical Institute
Ray, Frances, and Dale Cripps Scholarship Fund