Figure 1.

Bilateral targeting of BLA and vlOFC and confirmation of effective DREADD inhibition using ex vivo Ca²⁺ imaging in slices. A, Photomicrograph of hM4Di-mCherry DREADDs expression in BLA. Numerals indicate mm anterior to bregma. B, Reconstructions of viral expression of hM4Di (magenta) and eGFP (green) in BLA. The more intense colors represent regions where expression overlapped the most across animals. C, In BLA neurons expressing GCaMP6f and GCaMP6f + mCherry, application of CNO (10 µM) in the presence of picrotoxin (50 µM) had no effect on the frequency of elicited Ca²⁺ events (top left and right, example of single-cell traces; bottom left, Ca²⁺ event changes, each line is a cell). In BLA neurons that expressed hM4Di, there was a reduction in the frequency of elicited Ca²⁺ events during CNO application (bottom right). D, Photomicrograph of hM4Di-mCherry DREADDs expression in vlOFC. Numerals indicate mm anterior to Bregma. E, Reconstruction of viral expression of hM4Di (magenta) and eGFP (green). The more intense colors represent regions where expression overlapped the most across animals. F, In vlOFC neurons expressing GCaMP6f and GCaMP6f + mCherry, application of CNO (10 µM) had no effect on the frequency of elicited Ca²⁺ events (top left and right, example of single-cell traces; bottom left, Ca²⁺ event changes; each line is a cell). In vlOFC neurons that expressed hM4Di, there was a reduction in the frequency of Ca²⁺ events during CNO application (bottom right). n = 2–5 slices/rat, two-way ANOVA and multiple comparison tests. ***p < 0.001, *p < 0.05.