Pan, XiaopingDutta, Neesha Kishore2025-07-032025-052025-05-07May 2025http://hdl.handle.net/10342/14193Nicotine, a psychoactive alkaloid found in tobacco, drives addiction by binding to nicotinic acetylcholine receptors (nAChRs), altering cholinergic signaling, and promoting receptor overexpression. The overexpression of nAChRs has been linked to drug dependence, making nicotine addiction a leading cause of preventable deaths worldwide, largely due to tobacco use and secondhand smoke exposure. The nematode Caenorhabditis elegans, with its conserved nAChR subunits and well-characterized nervous system, exhibits behavioral responses to nicotine similar to those in mammals, including tolerance, acute stimulation, withdrawal, and sensitization. These traits allow for it to be an ideal model for studying the molecular and genetic mechanisms of nicotine addiction. The role of miRNAs in regulating nAChRs, nicotine-dependent behaviors, and the epigenetic effects involved in transgenerational inheritance will be explored in this study, with a focus on miRNA inhibition of cel-miR-100a to knock down miR-100a implicated in addiction pathways. Behavioral assays demonstrate nicotine dependence, withdrawal-like effects, and transgenerational inheritance, with F2-generation worms, which are never directly exposed to nicotine, but exhibit increased locomotion on nicotine-free plates. Experimental protocols begin with the cultivation of synchronized C. elegans populations (F0 generation) and chronic (24-hour) exposure to 61.7µM nicotine in K-medium. Behavioral assessments are performed using the WormLab software, revealing changes in addiction-related behaviors and suggesting miRNA regulation of nAChRs as a key molecular mechanism. Statistical analyses, such as ANOVA and regression modeling, are further used to evaluate gene expression and behavioral data. These findings provide novel insights into the molecular and genetic basis of nicotine addiction, including its transgenerational risks, and could inform the development of miRNA-based therapeutic methods to combat nicotine dependence and its long-term risks.application/pdfNicotine dependencemiRNA regulationTransgenerational inheritanceHonors Thesis2025-06-12