NDLI: Novel whole-tissue quantitative assay of nitric oxide levels in Drosophila neuroinflammatory response.

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Year: 2013 Issue: 82

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Novel whole-tissue quantitative assay of nitric oxide levels in Drosophila neuroinflammatory response.

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Novel whole-tissue quantitative assay of nitric oxide levels in Drosophila neuroinflammatory response.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Ajjuri, Rami R. O'Donnell, Janis M.
Description	Author Affiliation: Ajjuri RR ( Department of Biological Sciences, University of Alabama.)
Abstract	Neuroinflammation is a complex innate immune response vital to the healthy function of the central nervous system (CNS). Under normal conditions, an intricate network of inducers, detectors, and activators rapidly responds to neuron damage, infection or other immune infractions. This inflammation of immune cells is intimately associated with the pathology of neurodegenerative disorders, such as Parkinson's disease (PD), Alzheimer's disease and ALS. Under compromised disease states, chronic inflammation, intended to minimize neuron damage, may lead to an over-excitation of the immune cells, ultimately resulting in the exacerbation of disease progression. For example, loss of dopaminergic neurons in the midbrain, a hallmark of PD, is accelerated by the excessive activation of the inflammatory response. Though the cause of PD is largely unknown, exposure to environmental toxins has been implicated in the onset of sporadic cases. The herbicide paraquat, for example, has been shown to induce Parkinsonian-like pathology in several animal models, including Drosophila melanogaster. Here, we have used the conserved innate immune response in Drosophila to develop an assay capable of detecting varying levels of nitric oxide, a cell-signaling molecule critical to the activation of the inflammatory response cascade and targeted neuron death. Using paraquat-induced neuronal damage, we assess the impact of these immune insults on neuroinflammatory stimulation through the use of a novel, quantitative assay. Whole brains are fully extracted from flies either exposed to neurotoxins or of genotypes that elevate susceptibility to neurodegeneration then incubated in cell-culture media. Then, using the principles of the Griess reagent reaction, we are able to detect minor changes in the secretion of nitric oxide into cell-culture media, essentially creating a primary live-tissue model in a simple procedure. The utility of this model is amplified by the robust genetic and molecular complexity of Drosophila melanogaster, and this assay can be modified to be applicable to other Drosophila tissues or even other small, whole-organism inflammation models.
File Format	HTM / HTML
e-ISSN	1940087X
DOI	10.3791/50892
Journal	Journal of Visualized Experiments
Issue Number	82
Language	English
Publisher	MyJove Corp.
Publisher Date	2013-12-04
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Discipline Physical Sciences Discipline Life Sciences Discipline Medicine Inflammation Metabolism Nitric Oxide Animals Brain Brain Chemistry Brain Diseases Drosophila Research Support, N.i.h., Extramural Video-audio Media
Content Type	Text
Resource Type	Article

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