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. 2020 Dec 5;13:100284. doi: 10.1016/j.ynstr.2020.100284

Fig. 3.

Fig. 3

A retrograde viral-genetic technique permits experimental manipulation of LC neurons innervating specific terminal fields. Rats received bilateral injections of CAV2-PRS-CreV5 in CeA (n = 8) or mPFC (n = 8) and bilateral injections of Cre-inducible AAV-DIO-hM3Dq-mCherry or AAV-DIO-hM4Di-mCherry in LC. Representative images of injection sites in CeA and mPFC are shown in A&B, respectively. Boxes in low power images show the locations of high power images underneath. Areas enclosed in dotted lines in high power images show cannula tracks in CeA and evidence of minor glial scarring in mPFC that resulted from viral injections. Representative images of transduced neurons projecting to CeA and mPFC are shown in C&D, respectively. (E) These manipulations result in similar numbers of transduced neurons within each population. Noradrenergic neurons outside of LC were not found to express mCherry after these injections. A representative image of noradrenergic neurons in A5 is shown in F. Whole-cell recordings show that an hM3Dq + neuron increased its firing rate and an hM4Di + neuron was silenced in response to bath application of 1 μM CNO, respectively. Neurons lacking hM3Dq and hM4Di were insensitive to CNO (G). Scale bars in C, D&F = 100 μm.