Caspase involvement in autophagy

Caspases are a family of cysteine proteases widely known as the principal mediators of the apoptotic cell death response, but considerably less so as the contributors to the regulation of pathways outside cellular demise. In regards to autophagy, the modulatory roles of caspases have only recently begun to be adequately described. In contrast to apoptosis, autophagy promotes cell survival by providing energy and nutrients through the lysosomal degradation of cytoplasmic constituents. Under basal conditions autophagy and apoptosis cross-regulate each other through an elaborate network of interconnections which also includes the interplay between autophagyrelated proteins (ATGs) and caspases. In this review we focus on the effects of this crosstalk at the cellular level, as we aim to concentrate the main observations from research conducted so far on the fine-tuning of autophagy by caspases. Several members of this protease-family have been found to directly interact with key ATGs involved in different tiers across the autophagic cascade. Therefore, we firstly outline the core mechanism of macroautophagy in brief. In an effort to emphasize the importance of the intricate cross-regulation of ATGs and caspases, we also present examples drawn from Drosophila and plant models regarding the contribution of autophagy to apoptotic cell death during normal development

Assays to monitor mitophagy in Drosophila

Autophagy is a central pathway utilized by many eukaryotic cells in order to recycle intracellular constituents, particularly under periods of nutrient scarcity or cellular damage. The process is evolutionary conserved from yeast to mammals and can be highly selective with regards to the contents that are targeted for degradation. The availability of Drosophila transgenic lines and fluorophore-labelled autophagic markers allows nowadays for the more effortless visualization of the process within cells. Herein, we provide two protocols to prepare Drosophila samples for confocal and transmission electron microscopy for in vivo monitoring of mitophagy; a specific type of autophagy for the clearance of damaged or superfluous mitochondria from cells

Investigating Non-selective Autophagy in Drosophila

Drosophila melanogaster is a popular model organism in molecular genetics and cell biology. Various Drosophila tissues have been successfully used for studying autophagy, and our knowledge about the genetic regulation of this process is constantly growing. It is important to use assays that distinguish between non-selective autophagy and the selective forms. Here we introduce a selection of proven methods, which, taking into account their limitations, are suitable to measure non-selective autophagy in Drosophila fat and other tissues