Figure 5.
pS6-IR localization in activated neurons in the MB-ARC and VMH. A, Wild-type animals were allowed to feed ad libitum or were fasted for 24 h before treatment with leptin (Lep; 5 mg/kg, ip) or vehicle (Veh) for 1 h before the animals were killed and processing for the immunofluorescent detection of c-Fos-IR (red) and pS6-IR (green). Top panels show representative images of merged images at each condition. Smaller panels beneath show digital zoom of each channel and merged images from the boxed area in the larger panels above. Graphs: Total numbers of pS6-IR, c-Fos-IR, and percent double-labeled neurons in the MB-ARC and VMH are plotted as mean ± sem (n ≥ 3 per condition; bars with different letters were significantly different by ANOVA, P < 0.05). B, Wild-type animals under the indicated ad libitum-fed (Ad Lib Fed) or overnight (∼16 h) conditions were treated with icv vehicle or rapamycin ((Rapa) 2 μg/μl) via indwelling catheters overnight and for additional 2 h before the animals were killed and processing for the immunofluorescent detection of pS6-IR (green) and c-Fos-IR (red). Representative images under each condition are shown. In addition to the MB-ARC pS6/c-Fos-IR that is the focus of this analysis, note some inflammatory induction of pS6-IR in the ependymal cells that line the ventricle, which is also blocked by treatment with rapamycin. 3V, Third ventricle.