title: Quantification of mitochondrial morphology in neurites of dopaminergic neurons using multiple parameters.
abstract: BACKGROUND
  Studies of mitochondrial morphology vary in techniques. Most use one morphological parameter while others describe mitochondria qualitatively. Because mitochondria are so dynamic, a single parameter does not capture the true state of the network and may lead to erroneous conclusions. Thus, a gestalt method of analysis is warranted.
  
  
  METHODS
  This work describes a method combining immunofluorescence assays with computerized image analysis to measure the mitochondrial morphology within neuritic projections of a specific population of neurons. Six parameters of mitochondrial morphology were examined utilizing ImageJ to analyze colocalized signals.
  
  
  RESULTS
  Using primary neuronal cultures from Drosophila, we tested mitochondrial morphology in neurites of dopaminergic (DA) neurons. We validate our model using mutants with known defects in mitochondrial morphology. Furthermore, we show a difference in mitochondrial morphology between cells treated as control or with a neurotoxin inducing PD (Parkinson's Disease in humans)-like pathology. We also show interactions between morphological parameters and experimental treatment.
  
  
  METHODS
  Our method is a significant improvement of previously described methods. Six morphometric parameters are quantified, providing a gestalt analysis of mitochondrial morphology. Also it can target specific populations of mitochondria using immunofluorescence assay and image analysis.
  
  
  CONCLUSIONS
  We found that our method adequately detects differences in mitochondrial morphology between treatment groups. We conclude that some parameters may be unique to a mutation or a disease state, and the relationship between parameters is altered by experimental treatment. We suggest at least four variables should be considered when using mitochondrial structure as an experimental endpoint.
authors.affiliation: Functional Pharmacology, Department of Neuroscience, Biomedicinska Centrum, Uppsala University, Husargatan 3, Box 593, 75124 Uppsala, Sweden. Electronic address: lyle.wiemerslage@neuro.uu.se.
authors.forename: Lyle
authors.initials: L
authors.lastname: Wiemerslage
authors.affiliation: Neuroscience Program, Department of Biological Sciences, Ohio University, Athens, OH 45701, United States. Electronic address: leed1@ohio.edu.
authors.forename: Daewoo
authors.initials: D
authors.lastname: Lee
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D000831:Animals, Newborn; D002478:Cells, Cultured; D003598:Cytoskeletal Proteins / Q000235:genetics / Q000378:metabolism; D059290:Dopaminergic Neurons / Q000648:ultrastructure*; D004330:Drosophila; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism; D004625:Embryo, Nonmammalian; D019204:GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D007091:Image Processing, Computer-Assisted; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D008854:Microscopy, Electron; D008928:Mitochondria / Q000378:metabolism / Q000648:ultrastructure*; D016501:Neurites / Q000378:metabolism / Q000648:ultrastructure*; D025341:Principal Component Analysis; D013223:Statistics as Topic
chemical_list: D003598:Cytoskeletal Proteins; D029721:Drosophila Proteins; D008565:Membrane Proteins; C527116:OPA1 protein, Drosophila; D049452:Green Fluorescent Proteins; C504426:DRP1 protein, Drosophila; D019204:GTP-Binding Proteins
keywords: Drosophila; Mitochondria; Parkinson; drp1; opa1