Grantee Research Project Results
Detection of Nitric Oxide in Mice by Electron Paramagnetic Resonance TechniquesEPA Grant Number: U915227
Title: Detection of Nitric Oxide in Mice by Electron Paramagnetic Resonance Techniques
Investigators: Fecker, Jesse A.
Institution: Dartmouth College
EPA Project Officer: Smith, Bernice
Project Period: November 1, 1997 through November 1, 2000
Project Amount: $102,000
RFA: STAR Graduate Fellowships (1997)
Research Category: Academic Fellowships , Biology/Life Sciences , Fellowship - Biochemistry
The objective of this research project is to study the chemical and biological properties of dithiocarbamates, complexes used in trapping nitric oxide in vivo, in order to determine nitric oxide levels by electron paramagnetic resonance (EPR). The ability to detect and quantify nitric oxide will lead to an understanding of the role nitric oxide plays in neurological diseases, including Huntington's disease.Approach:
Three types of mice were used in these studies; control mice, transgenic mice exhibiting symptoms of Huntington's Disease, and the non-transgenic littermates of the same species. All mice were injected with 500 mg/kg diethyldithiocarbamate and 100 mg/kg iron(II) sulfate approximately one1 hour before sacrifice. Mice were injected with 100 mg/kg 3-nitropropionic acid at several times in order to generate a time -course response. Upon sacrifice, the mice were quickly dissected, and tissues were frozen in liquid nitrogen (77 K). The frozen tissues were analyzed by X-band EPR spectroscopy. Following spectroscopy, the amplitude of the signals attributed to copper and nitric oxide were measured from the EPR spectra. Through the use of standards containing known concentrations of copper and nitric oxide, it was possible to quantify the amount of copper and nitric oxide present in a tissue sample.Supplemental Keywords:
fellowship, nitric oxide, dithiocarbamate, electron paramagnetic resonance, (EPR), chelation, metal binding, free radicals, Huntington's Disease.