Caenorhabditis elegans chemosensory bioassay for seven transmembrane receptor ligands
This invention relates to methods for producing a transgenic C. elegans that expresses a human 7TMR pan-neuronally, such that said transgenic C. elegans exhibits a known phenotype. These transgenic C. elegans can be used in a variety of ways, including, but not limited to: (1) screening and identifying substances that bind to and activate particular human 7TMRs; (2) screening for substances that antagonize human 7TMR activation; (3) identifying human 7TMRs that may respond to particular substances; and (4) evaluating the specificity and efficacy of substances on human 7TMR activation.
1. A method for identifying at least one test substance that is an antagonist of a human 7 transmembrane receptor (h7TMR), said method comprising the steps of:
(a) providing at least one transgenic C. elegans whose genome comprises a nucleotide sequence encoding a h7TMR, wherein said h7TMR is expressed in sensory neurons of said transgenic C. elegans , and wherein said sensory neurons correlate with behavior, and wherein said transgenic C. elegans exhibits a phenotype;
(b) contacting said at least one transgenic C. elegans with at least one test substance, wherein said at least one test substance is distributed in a medium;
(c) determining whether said at least one test substance causes a suppression of said phenotype in said at least one transgenic C. elegans ; and
(d) identifying said at least one test substance that causes a suppression of said phenotype in said at least one transgenic C. elegans as an antagonist of said 7TMR.
2. The method as claimed in claim 1 , wherein said behavior is volatile chemoattraction, and said sensory neurons are AWA neurons.
3. The method as claimed in claim 1 , wherein said behavior is volatile chemorepulsion, and said sensory neurons are AWB neurons.
4. The method as claimed in claim 1 , wherein said behavior is water-soluble chemoattraction, and said sensory neurons are chosen from the group of: ASE, ADF, ASG, and ASI neurons.
5. The method as claimed in claim 1 , wherein said behavior is water-soluble chemorepulsion, and said sensory neurons are selected from the group consisting of: ASH and ADL neurons.
6. The method as claimed in claim 1 , wherein said behavior is dauer formation, and said sensory neurons are selected from the group consisting of: ASI, ASG, and ADF neurons.
7. The method as claimed in claim 1 , wherein said behavior is chosen from the group of: thermoattraction and thermorepulsion, and said sensory neurons are AFD neurons.
8. The method as claimed in claim 1 , wherein the medium is chosen from the group of: buffer, growth medium, and agar.
9. The method as claimed in claim 1 , wherein the medium is a growth medium that comprises a biomolecular separation in a matrix.
10. The method as claimed in claim 1 , wherein said matrix is chosen from the group of: agarose and polyacrylamide.