1. MITOCHONDRIAL COMPLEX 1 ACTIVITY IN MYOCARDIAL-SPECIFIC CHF1 DEFICIENT MICE Jeffrey Gordon 11/23/2010

2. CHF1 – A Brief Overview CHF1 is a basic-HLH transcription repressor which plays an important role in cardiovascular development CHF1 deficient mice are more susceptible to heart failure CHF1 myocardial transgenic mice are less susceptible to heart failure

3. My Project Hypothesis: differences of mitochondrial complex 1 activity may explain the phenotypic difference in susceptibility to heart failure in the myocardial specific CHF1 deficient mouse model Why investigate complex 1? Who cares / to what end?

4.  In heart failure, decreased mitochondrial content and overall mitochondrial dysfunction exist  There are increased RNA transcripts of complex 1 in CHF1 transgenic mice  Preliminary data suggests decreased mitochondrial respiration in CHF1 deficient mice  Complex I is a manageable target Why Test Complex 1 ?

5. Mitochondria  Mitochondria generate most of a cell’s ATP.  Mitochondria have a more broad function  Made of 4 complexes and ATP synthase http://en.wikipedia.org/wiki/File:ETC_electron_transport_chain.svg

6. Complex I  Largest complex – 45 different polypeptides  Redox reaction – NADH + H(+) + ubiquinone (Q) NAD(+) + dihydroubiquinone (QH2)

7. Who Cares / to what end?  Further characterize the mechanism responsible for increased susceptibility to heart failure of CHF1 deficient mice  Further understand mitochondria, which may offer therapeutic targets in the future

8. Overview of My Project 1. Breed wild-type controls and myocardial-specific CHF1 deficient mice 2. Isolate left ventricular mitochondrial tissue 3. Measure complex 1 activity via spectrophotometry 4. Normalize this to total mitochondrial activity 5. Compare the two groups

9. Spectrophotometry for Complex 1  Beer - Lambert Law  Measure light absorbance of a material to calculate concentration.  For complex 1: NADH + H(+) + ubiquinone (Q) NAD(+) + dihydroubiquinone (QH2) http://www.adinstruments.com/solutions/experiments/labchart_experiments/full.php ?exp_id=187&section_id=13&name_id=245&template=teaching

10. Results GenotypeC1 activityC1/CS F/F aMHC-Cre639.341.602 F/F aMHC-Cre567.731.836 F/F aMHC-Cre255.732.178 F/F aMHC-Cre506.351.61 F/F aMHC-Cre673.431.757 F/F427.931.932 F/F603.532.168 F/F499.542.055 F/F368.261.921 F/F334.161.802 F/F339.271.857 F/F521.72.3 F/F663.211.985 F/F579.241.428 -C1 = complex 1. CS = citrate synthase. -C1 Activity measured in nm/ (min *mg)

11. Results CHF1 Deficient WT P Value: 0.314564499

12. Conclusions  Complex 1 activity is not significantly different between wild-type and myocardial specific CHF- 1 deficient mice  Thus, differences in complex 1 are unlikely to explain the increased susceptibility to heart failure between these groups

13. Future Avenues  CHF1 deficient mice have abnormal with calcium signaling and apoptosis. Further studies into mitochondrial dysfunction may lead to better understandings of these mechanisms.