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Autophagy guide Autophagy Tools for cell death series Contents Introduction to cell death Autophagy -- Types of autophagy ?? Macroautophagy ?? Microautophagy ?? Chaperone ??mediated autophagy Autophagy cellular and molecular mechanisms -- Induction and ph

Autophagy Tools for cell death series Contents Introduction to cell death Autophagy -- Types of autophagy ?? Macroautophagy ?? Microautophagy ?? Chaperone ??mediated autophagy Autophagy cellular and molecular mechanisms -- Induction and phagophore formation -- Phagophore elongation and autophagosome formation -- Fusion degradation and recycling -- Autophagy regulation Autophagy and disease -- Cancer -- Neurodegenerative diseases -- Cardiovascular diseases -- Infectious disease Autophagy detection -- Markers of autophagy -- Visual identi ?cation -- ATG LC detection ?? Western Blotting ?? Turnover of LC ??II ATG ??PE ?? Luminescence based assays ?? GFP ??ATG LC detection ?? Fluorescence microscopy ?? Tandem mRFP mcherry ??GFP uorescence microscopy ?? Flow and multispectral imaging cytometry ?? Immunohistochemistry -- SQSTM p and related LC binding protein detection -- TOR AMPK and ATG ULK detection -- Additional autophagy related markers ?? ATG ?? ATG ?? ATG ?? BECLIN ?? DRAM General considerations for autophagy detection References Introduction to cell death Cell death happens when a biological cell fails to maintain essential life functions This may be the result of the natural process of development or protection against cellular defects of normal cells replacement or factors such as disease or localized injury Cell death can be classi ?ed according to its morphological appearance such as apoptotic or necrotic enzymological criteria with or without involvement of distinct proteases functional aspects programmed or accidental or immunological characteristics immunogenic or non ??immunogenic In this guide we want to examine the role of autophagy a self ??degradation process that has been historically linked to non ??apoptotic cell death Autophagy Autophagy from the Greek ??auto ? ??self and ??phagein ? ??to eat is a general term for processes by which dysfunctional cellular components are degraded inside the cell through the action of lysosomes Components of the cytoplasm including organelles are sequestered and moved into the lumen of lysosomes vacuoles where they are broken down into basic components and returned to the cytosol for reuse Under physiological conditions autophagy has a number of vital roles such as maintenance of the amino acid pool during starvation preimplantation development clearance of intracellular microbes and regulation of innate and adaptive immunity Autophagy has been widely implicated in many pathophysiological processes such as cancer metabolic and neurodegenerative disorders as well as cardiovascular and pulmonary diseases It also has an important role in aging and exercise One of the characteristic features of autophagy is its dynamic regulation cellular autophagic activity is usually low under basal conditions but there are a number of stimuli that can lead to its upregulation Nutrient starvation is the most well ??known inducer of autophagy but other biological stress stimuli such as hypoxia or energy depletion hormonal stimulation or pharmaceutical agents can increase autophagy in the cells as well Autophagy is a common morphological feature in dying cells and therefore it was presumed for a long time to be a cell death pathway Recent investigations show that one of its major functions is to work on keeping cells alive under stressful conditions that could