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J Vis Exp


Title:The Identification of Sea Lamprey Pheromones Using Bioassay-Guided Fractionation
Author(s):Scott AM; Li K; Li W;
Address:"Department of Fisheries and Wildlife, Michigan State University. Department of Fisheries and Wildlife, Michigan State University; liweim@msu.edu"
Journal Title:J Vis Exp
Year:2018
Volume:20180717
Issue:137
Page Number: -
DOI: 10.3791/58059
ISSN/ISBN:1940-087X (Electronic) 1940-087X (Linking)
Abstract:"Bioassay-guided fractionation is an iterative approach that uses the results of physiological and behavioral bioassays to guide the isolation and identification of an active pheromone compound. This method has resulted in the successful characterization of the chemical signals that function as pheromones in a wide range of animal species. Sea lampreys rely on olfaction to detect pheromones that mediate behavioral or physiological responses. We use this knowledge of fish biology to posit functions of putative pheromones and to guide the isolation and identification of active pheromone components. Chromatography is used to extract, concentrate, and separate compounds from the conditioned water. Electro-olfactogram (EOG) recordings are conducted to determine which fractions elicit olfactory responses. Two-choice maze behavioral assays are then used to determine if any of the odorous fractions are also behaviorally active and induce a preference. Spectrometric and spectroscopic methods provide the molecular weight and structural information to assist with the structure elucidation. The bioactivity of the pure compounds is confirmed with EOG and behavioral assays. The behavioral responses observed in the maze should ultimately be validated in a field setting to confirm their function in a natural stream setting. These bioassays play a dual role to 1) guide the fractionation process and 2) confirm and further define the bioactivity of isolated components. Here, we report the representative results of a sea lamprey pheromone identification that exemplify the utility of the bioassay-guided fractionation approach. The identification of sea lamprey pheromones is particularly important because a modulation of its pheromone communication system is among the options considered to control the invasive sea lamprey in the Laurentian Great Lakes. This method can be readily adapted to characterize the chemical communication in a broad array of taxa and shed light on waterborne chemical ecology"
Keywords:Animals Biological Assay/*methods Chemical Fractionation/*methods Petromyzon/*growth & development Pheromones/*chemistry;
Notes:"MedlineScott, Anne M Li, Ke Li, Weiming eng Research Support, Non-U.S. Gov't Video-Audio Media 2018/08/07 J Vis Exp. 2018 Jul 17; (137):58059. doi: 10.3791/58059"

 
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