1 research outputs found
Dual Electrochemical and Physiological Apoptosis Assay Detection of in Vivo Generated Nickel Chloride Induced DNA Damage in Caenorhabditis elegans
Environmental nickel exposure is
known to cause allergic reactions,
respiratory illness, and may be responsible for some forms of cancer
in humans. Nematodes are an excellent model organism to test for environmental
toxins, as they are prevalent in many different environments. Nickel
exposure has previously been shown to impact nematode life processes.
In this study, Caenorhabditis elegans nematodes exposed to NiCl<sub>2</sub> featured high levels of programmed
cell death (PCD) in a concentration-dependent manner as measured by
counting apoptotic corpses in the nematode germ line. A green fluorescent
protein (GFP) reporter transgene was used that highlights cell corpse
engulfment by fluorescence microscopy. Analysis of the reporter in
a <i>p53</i> mutant strain putatively indicates that the
PCDs are a result of genomic DNA damage. In order to assay the potential
genotoxic actions of NiCl<sub>2</sub>, DNA was extracted from nematodes
exposed to increasing concentrations of NiCl<sub>2</sub> and electrochemically
assayed. In vivo damaged DNA was immobilized on pyrolytic graphite
electrodes using the layer-by-layer (LbL) technique. Square-wave voltammograms
were obtained in the presence of redox mediator, ruthenium trisbipyridine
(RuÂ(bpy)<sub>3</sub><sup>2+</sup>), that catalytically oxidizes guanines
in DNA. Oxidative peak currents were shown to increase as a function
of NiCl<sub>2</sub> exposure, which further suggests that the extracted
DNA from nematodes exposed to the nickel was damaged. This report
demonstrates that our electrochemical biosensor can detect damage
at lower Ni concentrations than our physiological PCD assay and that
the results are predictive of physiological responses at higher concentrations.
Thus, a biological model for toxicity and animal disease can be assayed
using an electrochemical approach