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Resources » Paper

Hamilton, O. Scott et al. (2011) International Worm Meeting "WincG: a cyclic GMP biosensor for worms."

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  • Comments on Hamilton, O. Scott et al. (2011) International Worm Meeting "WincG: a cyclic GMP biosensor for worms." (0)

  • Overview

    Status:
    Publication type:
    Meeting_abstract
    WormBase ID:
    WBPaper00039804

    Hamilton, O. Scott, O'Halloran, Damien M., Brueggemann, Chantal, Juang, Bi-Tzen, & L'Etoile, Noelle D. (2011). WincG: a cyclic GMP biosensor for worms presented in International Worm Meeting. Unpublished information; cite only with author permission.

    Cyclic 3',5'-guanosine monophosphate (cGMP) is a second messenger that has many cellular functions including regulating ion channels and protein kinases. cGMP is catalyzed from guanosine triphosphate (GTP) by guanylyl cyclases and hydrolyzed by phosphodiesterases to guanosine monophosphate (GMP). Given the diversity of cellular functions mediated by cGMP, a means of monitoring and measuring in vivo cGMP dynamics in C. elegans would provide the worm community with better resolution to probe many cellular pathways. Within the past few years a number of cGMP biosensors have been developed for mammalian cell cultures including FRET based reporters and, more recently, a non-FRET based single-fluorophore reporter that combines a circularly permutated EGFP with the cGMP binding sites of the regulatory domain of the Bos taurus PKG I that the authors (Nausch et al., PNAS, 2008) named FlincG (Fluorescent indicator of cyclic GMP). Since there are currently no available cGMP biosensors for nematodes, we have expressed a modified version of the FlincG in C. elegans that we call WincG (Worm indicator of cyclic GMP). By cloning the FlincG into the worm expression vector pPD95.75, removing a non-coding region of the 3'UTR and enhancing the circularly permutated EGFP portion of the construct, we have been able to successfully express the cGMP biosensor under C. elegans promoters. Here we show that, as a proof of concept, when we express the WincG in phasmid neurons and introduce SDS as a repellent stimulus to the tail of a worm (that is immobilized in a worm trap) we are able to consistently measure cGMP changes (deltaF/F0%). We are also currently examining cGMP dynamics in amphid neurons.

    Affiliation:
    - University of California, Davis, Davis, CA.


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