Use of Liquid Chromatography Mass Spectrometry to Analyze UGT Activity on Sagebrush Xenobiotics
Abstract
Sagebrush (Artemisia tridentate), a common shrub of western North America, is highly adept at producing defensive toxic chemicals. This chemistry is thought to play a role in minimizing the rate of herbivore attacks. Despite the plant’s toxicity, some animals such as sage grouse (Centrocercus urophasianus) depend on sagebrush for shelter and up to 90 % of their nutrition over the winter. However, sage grouse are selective in choosing their dietary sagebrush. In order to understand sage-grouse preferences and proficiency at metabolizing certain kinds of sagebrush, we developed an in vitro assay to analyze the activity of UDP-glucuronosyltransferase (UGT) liver detoxification enzymes. UGT enzymes add a glucuronic acid group to xenobiotics through a condensation reaction. UGT from mouse liver was challenged with foliar extracts from two kinds of sagebrush; Early (Artemisia arbuscular ssp. longiloba) a sagebrush preferred by sage grouse and Wyoming (Artemisia tridentate ssp. wyomingensis) avoided by sage grouse. Liquid chromatography mass spectrometry (LCMS) was used to analyze the trend of elimination and production of substances from the two species of sagebrush. This experiment will be used to investigate the transformation of sagebrush compounds by sage-grouse liver UGTs as a way to understand healthy habitat restoration for sage grouse.
Use of Liquid Chromatography Mass Spectrometry to Analyze UGT Activity on Sagebrush Xenobiotics
Sagebrush (Artemisia tridentate), a common shrub of western North America, is highly adept at producing defensive toxic chemicals. This chemistry is thought to play a role in minimizing the rate of herbivore attacks. Despite the plant’s toxicity, some animals such as sage grouse (Centrocercus urophasianus) depend on sagebrush for shelter and up to 90 % of their nutrition over the winter. However, sage grouse are selective in choosing their dietary sagebrush. In order to understand sage-grouse preferences and proficiency at metabolizing certain kinds of sagebrush, we developed an in vitro assay to analyze the activity of UDP-glucuronosyltransferase (UGT) liver detoxification enzymes. UGT enzymes add a glucuronic acid group to xenobiotics through a condensation reaction. UGT from mouse liver was challenged with foliar extracts from two kinds of sagebrush; Early (Artemisia arbuscular ssp. longiloba) a sagebrush preferred by sage grouse and Wyoming (Artemisia tridentate ssp. wyomingensis) avoided by sage grouse. Liquid chromatography mass spectrometry (LCMS) was used to analyze the trend of elimination and production of substances from the two species of sagebrush. This experiment will be used to investigate the transformation of sagebrush compounds by sage-grouse liver UGTs as a way to understand healthy habitat restoration for sage grouse.