VA Ann Arbor Healthcare System

2 ARTICLES PUBLISHED IN JoVE

Medicine

Visualization of Endogenous Mitophagy Complexes In Situ in Human Pancreatic Beta Cells Utilizing Proximity Ligation Assay

Gemma Pearson¹, Scott A. Soleimanpour^1,2

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.

Biology

Complementary Approaches to Interrogate Mitophagy Flux in Pancreatic β-Cells

Elena Levi-D’Ancona^1,2, Vaibhav Sidarala¹, Scott A. Soleimanpour^1,3,4

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.