Quantitative Assessment of Pheromone-Induced Dauer Formation in Caenorhabditis elegans

Neal, Scott J.; Kim, Kyuhyung; Sengupta, Piali

doi:10.1007/978-1-62703-619-1_20

Cited by 20 publications

(20 citation statements)

References 28 publications

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“…Donor oligo sequences for rict-1 All examined strains were cultured for at least 3 generations without being 470 allowed to starve prior to use in dauer assays. Dauer formation assays were performed 471 essentially as described (90). In brief, 5 growth-synchronized young adult animals grown 472 on OP50 were placed on dauer assay agar plates (90) with (for pheromone-induced dauer 473 formation assays) or without (for heat-induced dauer formation assays) ascr#5 474 pheromone and live E. coli.…”

Section: Generation Of Near-isogenic Lines (Nils) 431mentioning

confidence: 99%

Rictor/TORC2 mediates gut-to-brain signaling in the regulation of phenotypic plasticity inC. elegans

O’Donnell

Chao

Kammenga

et al. 2017

Preprint

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1Animals integrate external cues with information about internal conditions such as 2 metabolic state to execute the appropriate behavioral and developmental decisions. 3Information about food quality and quantity is assessed by the intestine and transmitted to 4 modulate neuronal functions via mechanisms that are not fully understood. The 5 conserved Target of Rapamycin complex 2 (TORC2) controls multiple processes in 6 response to cellular stressors and growth factors. Here we show that TORC2 coordinates 7 larval development and adult behaviors in response to environmental cues and feeding 8 state in the bacterivorous nematode C. elegans. During development, pheromone, 9 bacterial food, and temperature regulate expression of the daf-7 TGF-β and daf-28 10 insulin-like peptide in sensory neurons to promote a binary decision between 11 reproductive growth and entry into the alternate dauer larval stage. We find that TORC2 12 acts in the intestine to regulate neuronal expression of both daf-7 and daf-28, which 13 together reflect bacterial-diet dependent feeding status, thus providing a mechanism for 14 integration of food signals with external cues in the regulation of neuroendocrine gene 15 expression. In the adult, TORC2 similarly acts in the intestine to modulate food-regulated 16 foraging behaviors via the PDFR-1 neuropeptide receptor. We also demonstrate that 17 genetic variation affects food-dependent larval and adult phenotypes, and identify 18 quantitative trait loci (QTL) associated with these traits. Together, these results suggest 19 that TORC2 acts as a hub for communication of feeding state information from the gut to 20 the brain, thereby contributing to modulation of neuronal function by internal state. 21

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Section: Generation Of Near-isogenic Lines (Nils) 431mentioning

confidence: 99%

Rictor/TORC2 mediates gut-to-brain signaling in the regulation of phenotypic plasticity inC. elegans

O’Donnell

Chao

Kammenga

et al. 2017

Preprint

Self Cite

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“…First, we subjected newly hatched larvae to high concentrations of crude dauer pheromone as previously described, and scored the percentage of animals that formed dauers (Neal et al 2013). As expected, wild-type larvae populations formed significantly more dauers in high pheromone conditions (82.2 6 2.4%) compared to control water plates (4.1 6 0.8%) ( Figure 1A).…”

Section: Endogenous Rnai Pathways Are Required For Dauer Formationmentioning

confidence: 60%

“…To test dauer formation in the presence of high pheromone concentrations, we conducted assays using crude pheromone as previously described with a few modifications (Neal et al 2013;Zhang et al 2013). For each independent batch of crude pheromone, one activity unit was defined as the amount of pheromone that resulted in 33% dauers in wild-type animals (Zhang et al 2013).…”

Section: Dauer Formation Assaysmentioning

confidence: 99%

Endogenous RNAi Pathways Are Required in Neurons for Dauer Formation in Caenorhabditis elegans

Bharadwaj¹,

Hall

2017

Genetics

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Animals can adapt to unfavorable environments through changes in physiology or behavior. In the nematode, Caenorhabditis elegans, environmental conditions perceived early in development determine whether the animal enters either the reproductive cycle, or enters into an alternative diapause stage named dauer. Here, we show that endogenous RNAi pathways play a role in dauer formation in crowding (high pheromone), starvation, and high temperature conditions. Disruption of the Mutator proteins or the nuclear Argonaute CSR-1 result in differential dauer-deficient phenotypes that are dependent upon the experienced environmental stress. We provide evidence that the RNAi pathways function in chemosensory neurons for dauer formation, upstream of the TGF-b and insulin signaling pathways. In addition, we show that Mutator MUT-16 expression in a subset of individual pheromone-sensing neurons is sufficient for dauer formation in high pheromone conditions, but not in starvation or high temperature conditions. Furthermore, we also show that MUT-16 and CSR-1 are required for expression of a subset of G proteins with functions in the detection of pheromone components. Together, our data suggest a model where Mutator-amplified siRNAs that associate with the CSR-1 pathway promote expression of genes required for the detection and signaling of environmental conditions to regulate development and behavior in C. elegans. This study highlights a mechanism whereby RNAi pathways mediate the link between environmental stress and adaptive phenotypic plasticity in animals.

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“…A video demonstrating their protocol is included in Schroeder and Flatt (2014). A detailed protocol for assaying dauer larva formation induced by crude dauer pheromone is also available (Neal et al, 2013). Alternatively, the potency of dauer pheromone can be assayed by testing for inhibition of dauer recovery (Golden and Riddle, 1984c).…”

Section: Methods To Induce Dauer Formationmentioning

confidence: 99%

Working with dauer larvae

Karp¹

2018

WormBook

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Quantitative Assessment of Pheromone-Induced Dauer Formation in Caenorhabditis elegans

Cited by 20 publications

References 28 publications

Rictor/TORC2 mediates gut-to-brain signaling in the regulation of phenotypic plasticity inC. elegans

Rictor/TORC2 mediates gut-to-brain signaling in the regulation of phenotypic plasticity inC. elegans

Endogenous RNAi Pathways Are Required in Neurons for Dauer Formation in Caenorhabditis elegans

Working with dauer larvae

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