Examination and quantification of hemosiderin deposits in the spleen of C57Bl/6J mice exposed to methamphetamine,

Abstract

Methamphetamine (METH), a widely used psychostimulant, is known to produce drastic changes to different system throughout the body by increasing vasodilation, blood pressure, and heart rate. The spleen is a multifunctional organ to include immune, hematopoietic, and filtering of effete red blood cells. Alterations in splenic function can indicate local and/or systemic toxicity. We noted qualitative changes in the spleens of mice exposed to chronic METH and include gross weight changes, hemosiderin deposition, and morphological alterations in red and white pulp when compared to saline controls. Overabundant iron has detrimental effects on the body via oxidative phosphorylation and mitochondrial dysfunction leading to cell death and organ damage. We will quantitate the observed iron increases in the spleen of 14 male C57Bl/6J mice given daily i.p. injections of 5mg/kg METH or saline for 10 days. The spleens were removed, homogenized, and dry ashed using heat to remove organic content. The total iron content will be determined using a colorimetric assay in which organic materials are burned off, iron dissolved in nitric acid, and concentrations obtained with fluorescence spectrometry. Future experiments will be conducted to examine iron deposits in other organs as well as to determine the mechanism by which deposition occurs.