VA Ann Arbor Healthcare System 3 articles published in JoVE Biology Complementary Approaches to Interrogate Mitophagy Flux in Pancreatic β-Cells Elena Levi-D’Ancona1,2, Vaibhav Sidarala1, Scott A. Soleimanpour1,3,4 1Department of Internal Medicine, Division of Metabolism, Endocrinology and Diabetes, University of Michigan, Ann Arbor, 2Graduate Program in Immunology, University of Michigan Medical School, 3Department of Molecular and Integrative Physiology, University of Michigan, Ann Arbor, 4VA Ann Arbor Healthcare System This protocol outlines two methods for the quantitative analysis of mitophagy in pancreatic β-cells: first, a combination of cell-permeable mitochondria-specific dyes, and second, a genetically encoded mitophagy reporter. These two techniques are complementary and can be deployed based on specific needs, allowing for flexibility and precision in quantitatively addressing mitochondrial quality control. Behavior Automated Rat Single-Pellet Reaching with 3-Dimensional Reconstruction of Paw and Digit Trajectories Alexandra Bova1, Krista Kernodle1, Kaitlyn Mulligan2, Daniel Leventhal2,3,4 1Neuroscience Graduate Program, University of Michigan, 2Department of Neurology, University of Michigan, 3Department of Biomedical Engineering, University of Michigan, 4Department of Neurology, VA Ann Arbor Health System Rodent skilled reaching is commonly used to study dexterous skills, but requires significant time and effort to implement the task and analyze the behavior. We describe an automated version of skilled reaching with motion tracking and three-dimensional reconstruction of reach trajectories. Medicine Visualization of Endogenous Mitophagy Complexes In Situ in Human Pancreatic Beta Cells Utilizing Proximity Ligation Assay Gemma Pearson1, Scott A. Soleimanpour1,2 1Department of Internal Medicine, Division of Metabolism, Endocrinology and Diabetes, University of Michigan, Ann Arbor, 2VA Ann Arbor Healthcare System This protocol outlines a method for quantitative analysis of mitophagy protein complex formation specifically in beta cells from primary human islet samples. This technique thus allows analysis of mitophagy from limited biological material, which are crucial in precious human pancreatic beta cell samples.
