EdU, a new thymidine analogue for labelling proliferating cells in the nervous system
Chehrehasa, Fatemeh, Meedeniya, Adrian C.B., Dwyer, Patrick, Abrahamsen, Greger, & Mackay-Sim, Alan (2009) EdU, a new thymidine analogue for labelling proliferating cells in the nervous system. Journal of Neuroscience Methods, 177(1), pp. 122-130.
The standard method of labelling proliferating cells uses the thymidine analogue, bromodeoxyuridine (BrdU), which incorporates into the DNA during S-phase of the cell cycle. A disadvantage of this method is that the immunochemical processing requires pre-treatment of the cells and tissue with heat or acid to reveal the antigen. This pre-treatment reduces reliability of the method and degrades the specimen, reducing the ability for multiple immuno-fluorescence labelling at high resolution. We report here the utility of a novel thymidine analogue, ethynyl deoxyuridine (EdU), detected with a fluorescent azide via the “click” chemistry reaction (the Huisgen 1,3-dipolar cycloaddition reaction of an organic azide to a terminal acetylene). The detection of EdU requires no heat or acid treatment and the incorporated EdU is covalently conjugated to fluorescent probe. The reaction is quick and compatible with fluorescence immunochemistry and other fluorescent probes. We show here that EdU is non-toxic in vitro and in vivo and can be used in place of BrdU to label cells during neurogenesis and the progeny identified at least 30 days later. The fluorescent labelling of EdU, markedly improves the detection of proliferating cells and allows concurrent high resolution fluorescence immunochemistry.
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|Item Type:||Journal Article|
|Keywords:||BrdU, EdU, proliferation, neurogenic, Brain|
|Subjects:||Australian and New Zealand Standard Research Classification > BIOLOGICAL SCIENCES (060000)|
|Divisions:||Current > QUT Faculties and Divisions > Faculty of Health
Current > Institutes > Institute of Health and Biomedical Innovation
|Copyright Owner:||Copyright 2009 Elsevier|
|Copyright Statement:||This is the author’s version of a work that was accepted for publication in Journal of Neuroscience Methods. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Journal of Neuroscience Methods, [VOL 177, ISSUE 1, (2009)] DOI: 10.1016/j.jneumeth.2008|
|Deposited On:||06 Apr 2014 23:15|
|Last Modified:||31 May 2014 03:42|
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