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