Tags: autophagy

The workflow enables the quantification of autophagosomes in Arabidopsis thaliana root epidermal cells in 3D. It combines immunolabeling of an autophagosome marker ATG8 with commercially available anti-ATG antibody and the subsequent stereological quantification of the immunolabeled particles. The immunolabeled samples are imaged with a confocal microscope and Z-stacks are acquired.

Macroautophagy is often quantified by live imaging of autophagosomes labeled with fluorescently tagged ATG8 protein (FP-ATG8) in Arabidopsis thaliana. The labeled particles are then counted in single focal planes. This approach may lead to inaccurate results as the actual 3D distribution of autophagosomes is not taken into account and appropriate sampling in the Z-direction is not performed.

Autophagy is an evolutionary conserved catabolic process deemed to maintain or restore cellular and organismal homeostasis. In plants, basal autophagy is essential for growth and development, it is required for nutrient remobilization during senescence and nutrient deficiency, for removal of organelles and macromolecules formed during plant development or damaged by environmental stresses.