Effects of Larval Density on Gene Regulation in Caenorhabditis elegans During Routine L1 Synchronization
UMass Chan AffiliationsDepartment of Biochemistry and Molecular Pharmacology
UMass Metabolic Network
Document TypeJournal Article
Biochemical Phenomena, Metabolism, and Nutrition
Cellular and Molecular Physiology
Genetics and Genomics
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AbstractBleaching gravid C. elegans followed by a short period of starvation of the L1 larvae is a routine method performed by worm researchers for generating synchronous populations for experiments. During the process of investigating dietary effects on gene regulation in L1 stage worms by single-worm RNA-Seq, we found that the density of resuspended L1 larvae affects expression of many mRNAs. Specifically, a number of genes related to metabolism and signalling are highly expressed in worms arrested at low density, but are repressed at higher arrest densities. We generated a GFP reporter strain based on one of the most density-dependent genes in our dataset - lips-15 - and confirmed that this reporter was expressed specifically in worms arrested at relatively low density. Finally, we show that conditioned media from high density L1 cultures was able to downregulate lips-15 even in L1 animals arrested at low density, and experiments using daf-22 mutant animals demonstrated that this effect is not mediated by the ascaroside family of signalling pheromones. Together, our data implicate a soluble signalling molecule in density sensing by L1 stage C. elegans, and provide guidance for design of experiments focused on early developmental gene regulation.
G3 (Bethesda). 2018 Mar 30. pii: g3.118.200056. doi: 10.1534/g3.118.200056. Link to article on publisher's site
Permanent Link to this Itemhttp://hdl.handle.net/20.500.14038/29255
RightsCopyright © 2018. G3 applies a Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) to all its published material, per http://www.g3journal.org/content/scope-and-publication-policies#copyright.
Except where otherwise noted, this item's license is described as Copyright © 2018. G3 applies a Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) to all its published material, per http://www.g3journal.org/content/scope-and-publication-policies#copyright.