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. Author manuscript; available in PMC: 2014 Nov 1.
Published in final edited form as: Nat Med. 2014 Apr 13;20(5):531–535. doi: 10.1038/nm.3513

Figure 4. REDD1 over-expression in the rat mPFC increases depressive and anxiety-related behaviors and decreases mTORC1 signaling.

Figure 4

(a) Top. Human REDD1 was subcloned into an AAV2 plasmid under transcriptional regulation of the CMV promoter (AAV2-REDD1-IRES-eYFP). AAV2-IRES-eYFP plasmid without the human REDD1 gene was used as the control. Bottom. Schematic of experiment. Representative photomicrographs of injection sites in the mPFC are shown to the right. (b) Left. Mean sucrose preference (± SEM) for control (n=7) and AAV-REDD1 infused animals (n=7). Center. Mean immobility time (± SEM) for control (n=6) and AAV-REDD1 injected (n=7) animals. Right. Mean latency to feed (± SEM) for control (n=6) and AAV-REDD1 injected (n=7) animals. (c) Left. Mean entries to the center in the open field (+/− SEM) for control (n=7) and AAV-REDD1 injected (n=7) animals. Right. Mean center duration (± SEM) in the open field for control (n=7) and AAV-REDD1 injected (n=7) animals. (d) Left. Mean number (± SEM) of open arm entries in the elevated plus maze for control (n=6) and AAV-REDD1 injected (n=7) animals. Center. Mean open arm duration (± SEM) in the elevated plus maze for control (n=6) and AAV-REDD1 injected (n=7) animals. Right. Mean number (± SEM) of closed arm entries in the elevated plus maze for control (n=6) and AAV-REDD1 injected (n=7) animals. (e) Mean (± SEM) percent co-localization of eYFP and phospho-S6 Ribosomal protein in control (n=5) and REDD1 virus-injected (n=6) rats. Right. Representative co-localization images. (f) Mean (± SEM) fold change of protein from mPFC microdissections from control (n=10) or AAV-REDD1 injected (n=10) rats. Representative blots are shown to the right. (g) Left Top. Representative images of a co-localized mPFC layer V pyramidal neurons positive for GFP and DiI. Left Bottom. Representative images of high magnification Z-stack projections of segments of layer V pyramidal cell primary apical dendrites. Right. Mean ± SEM of spine density from control (n=12) and AAV-REDD1 (n=12) groups. (h) Mean ± SEM of spine density of Filopodia (F), Long-Thin (LT), and Mushroom/Stubby (M/S) spines from control and AAV-REDD1 groups is depicted (F Ctrl n=11; F REDD1 n=12; LT Ctrl n=11; LT REDD1 n=11; M/S Ctrl n=12; M/S REDD1 n=11). (*) p < 0.05 relative to control.