In vitro methods to monitor autophagy in Lepidoptera.
Autophagy is attracting growing interest, especially in relation to increasing evidence of the importance of autophagic processes in animal development, as well as in human cancer progression. In holometabolous insects (i.e., that undergo four distinct life cycle stages, including embryo, larva, pupa and imago), such as flies, butterflies, bees and beetles, autophagy has been found to play a fundamental role in metamorphosis, and given the high degree of conservation of the genes and the basic mechanisms of autophagy, attention to these relatively simple models has increased significantly. Together with Drosophila, Lepidoptera larvae are among the most common invertebrate models in studies concerning the protective action of starvation-induced autophagy or the possible role of autophagy as a programmed cell death process. In this chapter, we provide experimental methods developed for, or applicable to, the study of the autophagic process in the IPLB-LdFB cell line derived from the fat body of the caterpillar of the gypsy moth, Lymantria dispar.
Tettamanti
Gianluca
G
Department of Structural and Functional Biology, University of Insubria, Varese, Italy.
Malagoli
Davide
D
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001343:Autophagy / Q000502:physiology*; D001681:Biological Assay / Q000379:methods*; D053148:Caspase 3 / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure; D003599:Cytoskeleton / Q000378:metabolism / Q000648:ultrastructure; D004247:DNA / Q000378:metabolism; D004586:Electrophoresis / Q000379:methods; D006801:Humans; D020287:In Situ Nick-End Labeling; D007915:Lepidoptera / Q000502:physiology*; D008247:Lysosomes / Q000378:metabolism / Q000648:ultrastructure; D046529:Microscopy, Electron, Transmission / Q000379:methods; D008928:Mitochondria / Q000378:metabolism / Q000648:ultrastructure; D017382:Reactive Oxygen Species / Q000378:metabolism
D017382:Reactive Oxygen Species; D000255:Adenosine Triphosphate; D004247:DNA; D053148:Caspase 3