title: Methods to Assess Mitochondrial Morphology in Mammalian Cells Mounting Autophagic or Mitophagic Responses.
abstract: It is widely acknowledged that mitochondria are highly active structures that rapidly respond to cellular and environmental perturbations by changing their shape, number, and distribution. Mitochondrial remodeling is a key component of diverse biological processes, ranging from cell cycle progression to autophagy. In this chapter, we describe different methodologies for the morphological study of the mitochondrial network. Instructions are given for the preparation of samples for fluorescent microscopy, based on genetically encoded strategies or the employment of synthetic fluorescent dyes. We also propose detailed protocols to analyze mitochondrial morphometric parameters from both three-dimensional and bidimensional datasets. Finally, we describe a protocol for the visualization and quantification of mitochondrial structures through electron microscopy.
authors.affiliation: Section of Pathology, Oncology and Experimental Biology, Laboratory for Technologies of Advanced Therapies (LTTA), University of Ferrara, Ferrara, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Marchi
authors.affiliation: Section of Pathology, Oncology and Experimental Biology, Laboratory for Technologies of Advanced Therapies (LTTA), University of Ferrara, Ferrara, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Bonora
authors.affiliation: Section of Pathology, Oncology and Experimental Biology, Laboratory for Technologies of Advanced Therapies (LTTA), University of Ferrara, Ferrara, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Patergnani
authors.affiliation: Section of Pathology, Oncology and Experimental Biology, Laboratory for Technologies of Advanced Therapies (LTTA), University of Ferrara, Ferrara, Italy.
authors.forename: C
authors.initials: C
authors.lastname: Giorgi
authors.affiliation: Section of Pathology, Oncology and Experimental Biology, Laboratory for Technologies of Advanced Therapies (LTTA), University of Ferrara, Ferrara, Italy. Electronic address: paolo.pinton@unife.it.
authors.forename: P
authors.initials: P
authors.lastname: Pinton
mesh_terms: D000818:Animals; D001343:Autophagy; D005456:Fluorescent Dyes / Q000032:analysis; D049452:Green Fluorescent Proteins / Q000032:analysis; D006801:Humans; D018613:Microscopy, Confocal / Q000379:methods*; D008854:Microscopy, Electron / Q000379:methods*; D008928:Mitochondria / Q000378:metabolism / Q000648:ultrastructure*; D063306:Mitophagy; D061848:Optical Imaging / Q000379:methods*
chemical_list: D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins
keywords: Autophagy; Biogenesis; Fission; Fusion; GFP; Mitochondria; Mitochondrial morphology; Mitochondrial network; Mitophagy