Single-nucleus RNA-seq of Drosophila Thorax Post Exercise Treatment: Pilot Study
Authors: Bre Minniefield; Nicole Riddle
Affiliation: University of Alabama at Birmingham
Keywords: t. other (Exercise ); b. metabolism
Physical activity in any form can significantly decrease the incidence of various diseases and benefit overall health. However, we are unable to recommend optimized exercise treatments for a specific individual’s health needs. Despite significant progress, we lack a thorough understanding of the biological pathways that are activated or impacted by exercise. For example, little is known about how individual tissues are affected by exercise treatments and which cell- and tissue-levels change in response to exercise. Additionally, little is known about the pathway(s) that mediate short- and long-term exercise responses. Identifying molecular mediators of exercise response has the potential to provide trackable factors to better predict the outcomes of exercise for an individual, thus, allowing for optimized and personalized exercise treatments.
Here, we use a Drosophila exercise model to gain an understanding of the cell- and tissue-level changes precipitated by a short-term exercise treatment. Rotational stimulation by rotating the fly enclosures has been shown to increase animal activity in Drosophila, modeling exercise. Depending on genotype, animals treated with rotational stimulation show increases in activity up to 10-fold, and this increased activity alters gene expression, metabolite levels, and body condition. Using our established protocols with the TreadWheel exercise system, here, we carry out a single-nucleus transcriptome study focused on muscle to compare exercise-treated and control animals. Specifically, we use a wild-derived strain, DGRP line 304, which in previous studies showed a significant change in activity level with rotation and a change in weight as response to exercise in both sexes. Exercise-treated animals experienced five consecutive days of rotational stimulation for 2hrs, while control animals do not. Post exercise, thorax dissections were performed, a body part which consists of ~95% muscle. Using 10xGenomics technology, we collect single-nucleus transcriptome profiles from this tissue. We use these profiles to identify expression patterns, key cell types, and molecular pathways that are 1) differing between exercised and controlled flies; and 2) significantly affected by exercise. The analysis is in progress, and results will be reported. Discovering the key elements of exercise response will provide the foundational data to develop effective, personalized exercise plans, including alternatives for individuals unable to exercise.