Figure 3
From: Deciphering and modulating G protein signalling in C. elegans using the DREADD technology
Pharmacological characterisation of ceGAR-3Dq.
Agonist specificity was determined using dynamic mass redistribution. (A) GAR-3b is stimulated by 100 μM CCh, whereas 10 μM CNO does not stimulate the receptor. (B) 100 μM CCh cannot stimulate ceCAR-3b, but the DREADD is stimulated by 10 μM CNO. (C) The DREADD agonist CNO activates ceGAR-3Dq in a concentration dependent manner whereas the muscarinic agonist CCh does not have an effect on receptor activity. Given are one of three representative experiments performed in triplicates. (D) Second messenger assays reveal Gq-protein coupling of GAR-3b and ceGAR-3Dq. Transfected cells were incubated with media (non-stimulated), 100 μM CCh, or 10 μM CNO. CCh-stimulation of GAR-3b leads to a robust increase in IP formation, but ceGAR-3Dq is only activated by CNO. Given is the mean ± SD of three to four independent experiments performed in triplicates. (E) Calcium release was measured in ceGAR-3Dq transfected cells after stimulation with CCh and CNO. While CCh does not trigger Calcium release, CNO results into a concentration-dependent Calcium release. (F) GAR-3b transfected cells release Calcium after stimulation with CCh but not CNO. Given is the mean ± SEM of three independent experiments performed in duplicates.
