Discipline: Biological Sciences
Subcategory: Physiology and Health
Destiny Mitchell - Mississippi Valley State University
Co-Author(s): Romain Harmancey, University of Mississippi Medical Center, Jackson, MS
Type 2 diabetes accounts for 95% of the 25.8 million diabetic cases in the United States, and projections anticipate that 1 in 3 Americans will suffer from the disease by 2050. Our laboratory has previously demonstrated that cardiac mitochondrial uncoupling protein (UCP) 3 levels are down-regulated by 50 to 60% in rodent models of insulin resistance and hyperinsulinemia. Other groups have also demonstrated that UCP3 is cardio protective in the setting of myocardial infarction and reperfusion. Our research investigates the molecular mechanisms by which UCP3 protects the heart in the presence of a heart attack. We hypothesize that decreased mitochondrial UCP3 content is directly responsible for the impairment of long-chain fatty acid oxidation at reperfusion and increased death rate of diabetic patients following a heart attack. The goal of this project is aimed towards making an adequate UCP3 deficient model to test this hypothesis. CRISPR/Cas 9 was used to generate two lines of rodent models bearing mutations in the UCP 3 locus. The information presented in this study investigates the genotyping by restriction fragment length analysis. In order to confirm the complete loss of UCP 3, western blots were performed in various tissue samples of the rodent models. The three genotypes that were introduced in the rodent models using the CRISPR/Cas 9 technique were heterozygous, wild type, and knockout. The results from the western blots illustrated that UCP 3 content was not found in the mitochondria of the rodent tissue samples. Future studies will aim to determine whether the loss of UCP3 leads to alterations in myocardial metabolism, and more specifically whether this leads to impaired long-chain fatty acid oxidation following myocardial infarction and reperfusion.
Funder Acknowledgement(s): NIH R25 HL121042, R00 HL112952
Faculty Advisor: Rachel Beecham, firstname.lastname@example.org
Role: I took part in performing the western blots for the mitochondria of the different tissues. I also was in charge of determining the genotypes of the rat models.