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Resources » Paper

Nika, Liberta et al. (2015) International Worm Meeting "Fluorescent beads are a versatile tool to distinguish dauer larvae and other stages."

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  • Comments on Nika, Liberta et al. (2015) International Worm Meeting "Fluorescent beads are a versatile tool to distinguish dauer larvae and other stages." (0)

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    Status:
    Publication type:
    Meeting_abstract
    WormBase ID:
    WBPaper00047703

    Nika, Liberta, Gibson, Taylor, Konkus, Rebecca, & Karp, Xantha (2015). Fluorescent beads are a versatile tool to distinguish dauer larvae and other stages presented in International Worm Meeting. Unpublished information; cite only with author permission.

    C. elegans life history is dependent on environmental cues transduced through several signaling pathways. In favorable environmental conditions C. elegans develops continuously through four larval stages before molting into the adult stage. In contrast, adverse environmental conditions promote entry into the stress resistant dauer larva stage immediately following the second larval molt. Dauer larvae possess distinct properties including cellular and developmental arrest, extended lifespan, and discontinued feeding. If environmental conditions improve, dauer larvae recover to post-dauer L3 larvae, which are developmentally identical to continuously developing L3 larvae. Studies of dauer larvae have seeded discoveries in diverse fields, ranging from aging to neurobiology. Many of these studies rely on the unambiguous identification of dauer larvae. The most useful method to identify dauer larvae would be simple, scalable, and feasible even in mutant backgrounds that are defective in some aspects of dauer morphogenesis. None of the current methods possess all of these qualities. Here, we describe such a method taking advantage of the inability of dauer larvae to feed. Fluorescent beads are added to the bacterial food source, and dauer larvae are identified by a lack of beads within their digestive tract. We describe how this assay can be used to isolate dauer larvae formed by any of three common methods: starved plates, exogenous pheromone, and dauer-constitutive mutations. Lack of beads correlates well with other known markers of dauer formation, including greatly reduced pumping rate, presence of dauer alae and SDS resistance. We find that using beads rather than SDS-resistance to identify dauer larvae enables the recovery of SDS-sensitive mutants, including cuticle mutants and certain mutants within the dauer formation pathway. Finally, lack of beads also identifies molting larvae, extending the utility of this assay beyond dauer larvae.

    Affiliation:
    - Biology, Central Michigan University, Mount Pleasant, MI


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