Working with dauer larvae

Karp, Xantha

doi:10.1895/wormbook.1.180.1

Cited by 62 publications

(88 citation statements)

References 70 publications

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“…However, our knowledge regarding how the dauer entry decision is made and how the decision is coordinately executed across different tissues is still limited (Androwski et al, 2017). First, it is difficult to identify L2d, the stage when environmental signals are integrated and the dauer-commitment decision is made, because of its lack of distinct features (Karp, 2018). Additionally, it can be labor-intensive to look for nondauer features in dauers that fail to coordinately remodel all of their tissues.…”

Section: Introductionmentioning

confidence: 99%

Genetic markers enable the verification and manipulation of the dauer entry decision

Shih

Lee

Sternberg

2019

Developmental Biology

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Phenotypic plasticity allows animals to survive in changing environments through the alteration of phenotypes or development. One of the best-studied examples of phenotypic plasticity is dauer larval development in the free-living roundworm Caenorhabditis elegans. When faced with hostile environments, C. elegans larvae can exit reproductive development and enter the stress-resistant and spore-like dauer larval stage. However, knowledge about how the dauer entry decision is made, and how the different tissues of the animal coordinate to execute transformation into dauer, is limited. This is because identifying animals that make the entry decision, or that fail to coordinately remodel their tissues during dauer development, is time-consuming and labor-intensive. Utilizing our previously reported RNA-seq of animals going through dauer or reproductive development (Lee et al., 2017), we have identified genetic markers for conveniently tracking and manipulating the dauer entry decision. These include col-183 (which tracks dauer fate in the hypodermis), ets-10 (neurons and intestine), nhr-246 (intestine and hypodermis), and F53F1.4 (reproductive fate in the hypodermis). Using condition shift experiments, we demonstrate that the dauer-specific fluorescent expression of the markers correspond to the commitment event of the dauer entry decision, and therefore label when the decision is made. We show that these markers can be used to manipulate the entry decision by driving the reproduction-promoting gene daf-9 under the control of the dauer-specific marker col-183, through which we could shift animals into nondauer development. We further demonstrate that the markers can be used to track tissue coordination during the decision. daf-9, daf-15, and daf-18 partial dauers exhibit incomplete expression of the ets-10 marker, with our results indicating that the same gene (e.g. daf-9 or daf-18) can affect dauer development differently in different tissues. Our findings provide molecular tools for studying phenotypic plasticity during a whole animal decision.

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Section: Introductionmentioning

confidence: 99%

Genetic markers enable the verification and manipulation of the dauer entry decision

Shih

Lee

Sternberg

2019

Developmental Biology

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“…Previous systems-level studies have analyzed the C. elegans transcriptome during molt into dauer, during dauer, and during recovery from dauer (22)(23)(24). However, the transcriptome of L2d larvae during the dauer-entry decision has not been characterized, likely because L2d larvae lack strong distinguishing traits that can be used to isolate them (25). Furthermore, dauer and reproductive development have not been compared under parallel growth conditions, a lack that has limited the discovery of genes that are differentially expressed in the two developments.…”

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confidence: 99%

FMRFamide-like peptides expand the behavioral repertoire of a densely connected nervous system

Lee

Shih

Schaedel

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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Animals, including humans, can adapt to environmental stress through phenotypic plasticity. The free-living nematode can adapt to harsh environments by undergoing a whole-animal change, involving exiting reproductive development and entering the stress-resistant dauer larval stage. The dauer is a dispersal stage with dauer-specific behaviors for finding and stowing onto carrier animals, but how dauers acquire these behaviors, despite having a physically limited nervous system of 302 neurons, is poorly understood. We compared dauer and reproductive development using whole-animal RNA sequencing at fine time points and at sufficient depth to measure transcriptional changes within single cells. We detected 8,042 genes differentially expressed during dauer and reproductive development and observed striking up-regulation of neuropeptide genes during dauer entry. We knocked down neuropeptide processing using mutants and demonstrate that neuropeptide signaling promotes the decision to enter dauer rather than reproductive development. We also demonstrate that during dauer neuropeptides modulate the dauer-specific nictation behavior (carrier animal-hitchhiking) and are necessary for switching from repulsion to CO (a carrier animal cue) in nondauers to CO attraction in dauers. We tested individual neuropeptides using CRISPR knockouts and existing strains and demonstrate that the combined effects of and mimic the effects of on nictation and CO attraction. Through meta-analysis, we discovered similar up-regulation of neuropeptides in the dauer-like infective juveniles of diverse parasitic nematodes, suggesting the antiparasitic target potential of SBT-1. Our findings reveal that, under stress, increased neuropeptide signaling in enhances their decision-making accuracy and expands their behavioral repertoire.

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“…The eggs were incubated at 25 C on standard NGM plates containing E. coli [64]. Dauer larvae induced using daf-c mutations were identified by SDS selection: incubation in 1% SDS for at least 10 minutes kills all non-dauer stages except for eggs, from which dauer larvae can easily be distinguished [65,66]. Dauers induced by starvation were identified based on dauer morphology: pharyngeal constriction, radial constriction, and the presence of dauer alae.…”

Section: Methods Detailsmentioning

confidence: 99%

“…Crosses were initiated with twelve L4 loxP>daf-18(+)>loxP; daf-18(0) and four L4 daf-18(0); p::Cre(opti) hermaphrodites at 20 C. The parent crosses were moved to new plates periodically over 2-3 days and each egg lay was shifted to 25 C once the parents were removed. Scoring tissue-specific excision mosaic dauer larvae Dauers were selected by incubation in 1% SDS and scored 65-100 hours after egg lay, which corresponds to 1-2 days after dauer formation [66]. Only hermaphrodite dauers were scored.…”

Section: Daf-18 Tissue-specific Rescue Experimentsmentioning

confidence: 99%

Cell Non-autonomous Function of daf-18/PTEN in the Somatic Gonad Coordinates Somatic Gonad and Germline Development in C. elegans Dauer Larvae

Tenen

Greenwald

2019

Current Biology

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Working with dauer larvae

Cited by 62 publications

References 70 publications

Genetic markers enable the verification and manipulation of the dauer entry decision

Genetic markers enable the verification and manipulation of the dauer entry decision

FMRFamide-like peptides expand the behavioral repertoire of a densely connected nervous system

Cell Non-autonomous Function of daf-18/PTEN in the Somatic Gonad Coordinates Somatic Gonad and Germline Development in C. elegans Dauer Larvae

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