Measuring PDE4A and PDE4B in Normal and Diseased Mouse Leg Muscle Jessica Hall & Brittany Matthews 1 June 2001

Duchenne’s Muscular Dystrophy  Genetic disorder involving muscle weakness  Early onset: 2-6 years  Symptoms  Named for its discoverer  Rarely affects females

What are PDE4A & PDE4B?  Enzymes involved in regulating energy  Help adrenaline control muscle function  Help insulin control sugar use in muscle

Gel Electrophoresis  Load samples onto gel  Move the sample through the gel

Western Blotting  Transfer the proteins onto the membrane  Add primary antibody  Add secondary antibody  Add substrate to membrane  Expose film

Initial Results  No bands on film  Tested the enzyme & substrate  Tested multiple primary antibodies  Tested commercial gels vs. homemade gels  Tested amount of sample loaded*  Tested incubation time for the primary antibody*

Results with PDE4A Antibody

Results with PDE4 Antibody

Conclusions  There is no difference between diseased and normal mice with PDE4A in leg muscle.  Diseased muscle has more of an unknown PDE4 protein, based on a non-selective PDE4 antibody.