Transmission electron microscopy protocol for characterising autophagy-dependent cell death (ADCD) in Drosophila...

Autophagy-dependent cell death (ADCD) describes a form of regulated cell death (RCD) that is driven by autophagy machinery. While other modes of RCD such as apoptosis and necrosis can be defined by distinct cellular features, the characterisation of ADCD relies primarily on genetic manipulation, with limited guidelines for assessing cellular morphology. Drosophila melanogaster larval midgut histolysis provides the strongest in vivo evidence of developmental ADCD, resulting in degradation of gastric caeca and the proventriculus during larval-to-pupal transition (metamorphosis). To establish morphological hallmarks of ADCD, in this protocol, we describe methods for dissecting and preparing larval midguts for morphological characterisation of ADCD using transmission electron microscopy (TEM).