Detection of genetically altered copper levels in Drosophila tissues by synchrotron x-ray fluorescence microscopy.

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...

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Main Authors: Jessica C Lye, Joab E C Hwang, David Paterson, Martin D de Jonge, Daryl L Howard, Richard Burke
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2011-01-01
Series:PLoS ONE
Online Access:http://europepmc.org/articles/PMC3203902?pdf=render
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author Jessica C Lye
Joab E C Hwang
David Paterson
Martin D de Jonge
Daryl L Howard
Richard Burke
author_facet Jessica C Lye
Joab E C Hwang
David Paterson
Martin D de Jonge
Daryl L Howard
Richard Burke
author_sort Jessica C Lye
collection DOAJ
description 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.
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spelling doaj.art-409ea223d19947e2accd08e59ded28a12022-12-22T02:42:52ZengPublic Library of Science (PLoS)PLoS ONE1932-62032011-01-01610e2686710.1371/journal.pone.0026867Detection of genetically altered copper levels in Drosophila tissues by synchrotron x-ray fluorescence microscopy.Jessica C LyeJoab E C HwangDavid PatersonMartin D de JongeDaryl L HowardRichard BurkeTissue-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.http://europepmc.org/articles/PMC3203902?pdf=render
spellingShingle Jessica C Lye
Joab E C Hwang
David Paterson
Martin D de Jonge
Daryl L Howard
Richard Burke
Detection of genetically altered copper levels in Drosophila tissues by synchrotron x-ray fluorescence microscopy.
PLoS ONE
title Detection of genetically altered copper levels in Drosophila tissues by synchrotron x-ray fluorescence microscopy.
title_full Detection of genetically altered copper levels in Drosophila tissues by synchrotron x-ray fluorescence microscopy.
title_fullStr Detection of genetically altered copper levels in Drosophila tissues by synchrotron x-ray fluorescence microscopy.
title_full_unstemmed Detection of genetically altered copper levels in Drosophila tissues by synchrotron x-ray fluorescence microscopy.
title_short Detection of genetically altered copper levels in Drosophila tissues by synchrotron x-ray fluorescence microscopy.
title_sort detection of genetically altered copper levels in drosophila tissues by synchrotron x ray fluorescence microscopy
url http://europepmc.org/articles/PMC3203902?pdf=render
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