title: Detection of genetically altered copper levels in Drosophila tissues by synchrotron x-ray fluorescence microscopy.
abstract: Tissue-specific manipulation of known copper transport genes in Drosophila tissues results in phenotypes that are presumably due to an alteration in copper levels in the targeted cells. However direct confirmation of this has to date been technically challenging. Measures of cellular copper content such as expression levels of copper-responsive genes or cuproenzyme activity levels, while useful, are indirect. First-generation copper-sensitive fluorophores show promise but currently lack the sensitivity required to detect subtle changes in copper levels. Moreover such techniques do not provide information regarding other relevant biometals such as zinc or iron. Traditional techniques for measuring elemental composition such as inductively coupled plasma mass spectroscopy are not sensitive enough for use with the small tissue amounts available in Drosophila research. Here we present synchrotron x-ray fluorescence microscopy analysis of two different Drosophila tissues, the larval wing imaginal disc, and sectioned adult fly heads and show that this technique can be used to detect changes in tissue copper levels caused by targeted manipulation of known copper homeostasis genes.
authors: School of Biological Sciences, Monash University, Melbourne, Victoria, Australia.
authors: Jessica C
authors: JC
authors: Lye
authors: Joab E C
authors: JE
authors: Hwang
authors: David
authors: D
authors: Paterson
authors: Martin D
authors: MD
authors: de Jonge
authors: Daryl L
authors: DL
authors: Howard
authors: Richard
authors: R
authors: Burke
mesh_terms: D000818:Animals; D001692:Biological Transport / Q000235:genetics; D001921:Brain / Q000378:metabolism; D003300:Copper / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000235:genetics* / Q000378:metabolism*; D005123:Eye / Q000378:metabolism; D017344:Genes, Insect / Q000235:genetics; D006257:Head; D060227:Imaginal Discs / Q000378:metabolism; D008856:Microscopy, Fluorescence / Q000379:methods*; D009928:Organ Specificity; D015203:Reproducibility of Results; D017356:Synchrotrons; D014921:Wings, Animal / Q000378:metabolism; D014965:X-Rays
chemical_list: D003300:Copper