Undergraduate Biotechnology Research Fellowship in Toxicology
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Overview
abstract
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The model organism Caenorhabditis elegans (C.elegans) has been widely used to decipher action mechanisms of toxicants/drugs and represent a promising non-vertebrate model for toxicological/pharmacological researches. Although progress has been made to understand the addiction mechanisms of many abused drugs including nicotine, how genes interact with environments and generate inheritable neurological and behavioral alternations has been rarely studied. In this project, we will investigate the nicotine-induced neurological behaviors in C.elegans using a multigenerational approach. Previously our study has characterized several addictive behavioral patterns following nicotine exposures in the P0 generation, including stimulation, withdrawal, and tolerance. In this project we will investigate the multigenerational impacts of nicotine on behavioral and gene expressions in the F1 and F2 generations. C. elegans will be treated with control, 1ppm (6.17?M) and 10ppm (61.7?M) of nicotine for 24-hour and the nicotine?dependent behaviors will be characterized using a worm-tracking system. As the primary target of nicotine is the nicotinic acetylcholine receptors (nAChRs), we will also detect the expression of 28 nAChRs genes and related microRNA genes which may regulate the genetic adaption. Methods: 1) C.elegans P0 generation will be treated with nicotine and total RNA will be extract from P0, F1, and F2 generations; 2) Design primers and measure expression of the 28 nAChR genes using the quantitative Real-time PCR; 3) Identify conserved miRNAs that potentially target nAChRs using a target prediction algorithm TargetScan; and 4) Analyze selected targeting microRNAs using a stem-loop real-time PCR procedure. The C. elegans genome encodes 28 nAChR genes that are highly conserved across species including humans. This project will test a novel and significant hypothesis: multigenerational behaviors are inherited and regulated by microRNA-directed gene expression of nicotine targets. This project will have a substantial impact on current body of knowledge on microRNA regulated epigenetic mechanisms in multigenerational nicotine effects.
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