title: Studying Mitochondrial Structure and Function in Drosophila Ovaries.
abstract: Analysis of the mitochondrial structure-function relationship is required for a thorough understanding of the regulatory mechanisms of mitochondrial functionality. Fluorescence microscopy is an indispensable tool for the direct assessment of mitochondrial structure and function in live cells and for studying the mitochondrial structure-function relationship, which is primarily modulated by the molecules governing fission and fusion events between mitochondria. This paper describes and demonstrates specific methods for studying mitochondrial structure and function in live as well as in fixed tissue in the model organism Drosophila melanogaster. The tissue of choice here is the Drosophila ovary, which can be isolated and made amenable for ex vivo live confocal microscopy. Furthermore, the paper describes how to genetically manipulate the mitochondrial fission protein, Drp1, in Drosophila ovaries to study the involvement of Drp1-driven mitochondrial fission in modulating the mitochondrial structure-function relationship. The broad use of such methods is demonstrated in already-published as well as in novel data. The described methods can be further extended towards understanding the direct impact of nutrients and/or growth factors on the mitochondrial properties ex vivo. Given that mitochondrial dysregulation underlies the etiology of various diseases, the described innovative methods developed in a genetically tractable model organism, Drosophila, are anticipated to contribute significantly to the understanding of the mechanistic details of the mitochondrial structure-function relationship and to the development of mitochondria-directed therapeutic strategies.
authors.affiliation: Department of Genetics, School of Medicine, University of Alabama at Birmingham.
authors.forename: Danitra J
authors.initials: DJ
authors.lastname: Parker
authors.affiliation: Department of Genetics, School of Medicine, University of Alabama at Birmingham.
authors.forename: Aida
authors.initials: A
authors.lastname: Moran
authors.affiliation: Department of Genetics, School of Medicine, University of Alabama at Birmingham; kasturi@uab.edu.
authors.forename: Kasturi
authors.initials: K
authors.lastname: Mitra
mesh_terms: D000818:Animals; D019927:Cyclin E / Q000378:metabolism; D003598:Cytoskeletal Proteins / Q000172:deficiency / Q000235:genetics / Q000378:metabolism; D004330:Drosophila / Q000378:metabolism*; D005260:Female; D019204:GTP-Binding Proteins / Q000172:deficiency / Q000235:genetics / Q000378:metabolism; D018613:Microscopy, Confocal; D008928:Mitochondria / Q000737:chemistry / Q000378:metabolism*; D063154:Mitochondrial Dynamics; D024101:Mitochondrial Proteins / Q000378:metabolism; D010053:Ovary / Q000378:metabolism; D038761:Photobleaching
chemical_list: D019927:Cyclin E; D003598:Cytoskeletal Proteins; D024101:Mitochondrial Proteins; C504426:DRP1 protein, Drosophila; D019204:GTP-Binding Proteins