autophagosome

By Christina Towers, PhD.

By Christina Towers, PhD.

By Christina Towers, PhD

Autophagy is an essential process that cells utilize to degrade and recycle damaged material and fuel metabolism, especially under stress.  The process is evolutionarily conserved and complex, relying on over 20 key proteins. Induction of autophagy is mediated by the formation of the ULK and BECLIN complexes, the latter of which includes BH3-containing proteins and AMBRA1, resulting in the formation of a double membrane phagophore structure.

Autophagy more than a cytosolic event

Autophagy is a cellular process whereby cytosolic components are broken down and eliminated or recycled. As a homeostatic mechanism, basal autophagic activity eliminates excess or abnormal proteins and organelles1. As an induced process, autophagy may be triggered by various external challenges, such as decreased nutrient and energy resources, and oxidative stress¹.

Apoptosis is a form of programmed cell death which is mediated by cysteine proteases called caspases. It is an essential phenomenon in the maintenance of homeostasis and growth of tissues, and it also plays a critical role in immune response. The cytomorphological alterations and the key features of apoptosis are listed below:

The human form of microtubule-associated protein light chain 3 (LC3) is expressed as 3 splice variants LC3A, LC3B, and LC3C.¹ LC3B is a subunit of the MAP1A and MAP1B microtubule-binding proteins and plays a central role in autophagosome membrane structure.

Autophagy is a conserved mechanism whereby cells form double membrane autophagosomes to sequester cytoplasmic components for subsequent destruction by fusion with lysosomes (eukaryotes) or vacuoles (yeast). Targets of autophagy include aging proteins, damaged organelles and invasive pathogens, and the resulting breakdown products can be recycled back to the cytoplasm for re-use under conditions of starvation (1).

Microtubule-associated protein light chain 3 (LC3) is considered one of the definitive markers of autophagy, and its use is widespread in labs throughout the world. Despite its popularity, there are several considerations when employing LC3 antibodies in immunoassays, Western blots in particular.