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G3 (Bethesda)

Title:		Effects of Larval Density on Gene Regulation in Caenorhabditis elegans During Routine L1 Synchronization
Author(s):		Chan IL; Rando OJ; Conine CC;
Address:		"Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA 01605. Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA 01605 Oliver.Rando@umassmed.edu Colin.Conine@umassmed.edu"
Journal Title:		G3 (Bethesda)
Year:		2018
Volume:		20180504
Issue:		5
Page Number:		1787 - 1793
DOI:		10.1534/g3.118.200056
ISSN/ISBN:		2160-1836 (Electronic) 2160-1836 (Linking)
Abstract:		"Bleaching 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 signaling 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 signaling pheromones. Together, our data implicate a soluble signaling molecule in density sensing by L1 stage C. elegans, and provide guidance for design of experiments focused on early developmental gene regulation"
Keywords:		"Animals Caenorhabditis elegans/*genetics/*growth & development *Gene Expression Regulation, Developmental Genes, Reporter Green Fluorescent Proteins/metabolism Larva/genetics Signal Transduction Solubility chemical signaling density effects development ge;"
Notes:		"MedlineChan, Io Long Rando, Oliver J Conine, Colin C eng R01 HD080224/HD/NICHD NIH HHS/ P40 OD010440/OD/NIH HHS/ Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't England 2018/04/01 G3 (Bethesda). 2018 May 4; 8(5):1787-1793. doi: 10.1534/g3.118.200056"