Extended Data Fig. 5: Physiological evidence for D2R mediating the action of dopamine on the CDNs.
From: Behavioral devaluation by local resistance to dopamine
(A) Example ROIs of the CDN axons used for our intensity and fluorescence lifetime imaging experiments. The CDNs project bilaterally, we only use one ROI on one side per animal for our experiments. (B) In the absence of ChR2-XXM driven stimulation, the effects of dopamine on CDN calcium are less obvious (n = 10 each). (C) Example experiment where ChR2-XXM is used to elevate CDN activity before bath applying dopamine (left: average trace of raw photon counts displayed as mean ± SEM, middle: normalized individual traces, right: normalized average trace displayed as mean ± SEM). Because the initial rise in calcium is variable, we wait until calcium levels stabilize before perfusing and use the 10 s immediately preceding the start of perfusion as a baseline. Traces in the main figures have been cropped to only show activity once calcium levels have stabilized. Control animals without perfusion are shown (n = 10 each). (D) Full trace of Fig. 2f, left panel: CDN calcium signal is depressed upon bath application of 100 µM dopamine (n = 10 each). (E) Whole animal D2R mutants do not show a depression in calcium signaling following bath application of 100 µM dopamine (n = 10 each). (F) Full trace of Fig. 2f, middle panel: animals with the D2R knockdown in the CDNs do not show depression of CDN calcium levels following bath application of 100 µM dopamine (n = 10 each). (G) Animals with the D2R knocked down with a second, independent RNAi in the CDNs do not show depression of CDN calcium levels following bath application of 100 µM dopamine (n = 10). (H) In the absence of ChR2-XXM driven stimulation dopamine application does not alter CaMKII activity in the CDNs (n = 10 each). (I) D2R mutants show a weak increase in CaMKII activity following bath application of 10 µM dopamine (n = 10 each). (J) Animals with the D2R knocked down with a second, independent RNAi in the CDNs show a weak increase in CaMKII activity following bath application of 10 µM dopamine (n = 10 each).
