Methods to Study the Mitochondria - qRT-PCR, Western Blotting, Immunofluorescence, Electron Microscopy, “Fluorescence”, and Seahorse – techniques used to study the effects of age-related diseases like Alzheimer’s

Various laboratories across the world have developed methods to study mitochondrial proteins/markers through extractions of mitochondrial RNA, protein, mitophagy/autophagy in Alzheimer’s disease (AD) and other age-related diseases. Techniques such as qRT-PCR, western blotting, immunofluorescence, transmission electron microscopy and Seahorse bioanalyzer, mitochondrial membrane potential, detection of mitophagy, and mitochondrial functional assays have been used as outlined in this article. Most of these techniques were performed in vitro (human and mouse neuronal cell lines, transfected with mutant APP or Tau cDNAs) and in vivo (brain tissues from different mouse models of Alzheimer’s and other neurological diseases). Mitochondrial abnormalities in Alzheimer’s disease reported various forms, such as excessive reactive oxygen species (ROS) production, mitochondrial calcium dyshomeostasis, loss of ATP, and defects in mitochondrial dynamics and transport, and mitophagy. Mitochondrial dysfunction is largely involved in aging, age-related diseases such as cancer, diabetes, obesity and neurodegenerative disorders, including Alzheimer’s disease (AD), Parkinson’s disease (PD), Huntington’s disease, amyotrophic lateral sclerosis (ALS), multiple sclerosis (MS) and others. The essence of our article is to update protocols/methods and make them available to students, scholars, and researchers of mitochondria. We believe our article is important and useful to future mitochondrial studies.