A review article published in Traffic by Maurizio Molinari’s laboratory describes state-of-the art tools and methods to monitor intracellular traffic of lysosomal enzymes, as well as the catabolic processes of autophagy and heterophagy.
Bellinzona, November 07, 2024 – Lysosomal compartments control the clearance of cell-own material (autophagy) and of material that cells endocytose from the external environment (heterophagy) to warrant supply of nutrients, to eliminate macromolecules or parts of organelles present in excess, aged, or containing toxic material. Defective cargo delivery to lysosomal compartments is harmful to cells and organs since it causes accumulation of toxic compounds and defective organellar homeostasis. Moreover, inherited or sporadic mutations in lysosomal proteins and enzymes may hamper their folding in the endoplasmic reticulum (ER) and their lysosomal transport via the Golgi compartment, resulting in a series of debilitating diseases defined as lysosomal storage disorders.
The published review describes tools and methods available to researchers to monitor intracellular transport events to endo-lysosomal compartments as well as autophagic and heterophagic fluxes to degradative organelles. It has been provided an up-to-date description of available single- and tandem-fluorescent reporters, mono- and multi-modal microscopy techniques, and artificial-intelligence analysis strategies. Tools developed in the last couple of years in our lab are also described. For example the tandem Halo-GFP reporter to monitor intracellular trafficking events in time, by light and electron microscopy, biochemistry, and flow cytometry, and LysoQuant, a deep-learning approach for the segmentation and classification of fluorescence images capturing multi parameters including size, distribution, number of acidic degradative compartments and cargo delivery within them.
Photo (from left): Mikhail Rudinskiy and Diego Morone
