After confirming that the GK gene was preferently knocked down in tanycytes cells, we sought to investigate whether it caused any alteration in the amount of food intake. At the top of the Fig. 6 the experimental procedure is showing. As shown in Fig. 6A, GK knockdown animals showed a significant increase in food intake over 12 h compared to both control groups. Specifically, GK knockdown animals ate 16 g of food pellet, whereas AdshβGal-EGFP-injected animals ate 11 g; unoperated animals ate 10 g. No significant differences were detected when comparing both control groups (unoperated and Ad-shβGal-EGFP).Figure 6Effect of GK knockdown on feeding behavior in control and GK knockdown animals. (A–F) Food intake (A), first meal duration (B), frequency of eating events (C), cumulative eating time (D), intervals and interval duration (F) were evaluated in unoperated, Ad-EGFP-injected (Ad-EGFP) and Ad-shGK-injected animals (Ad-shGK). Schematic representation of the experimental procedure is shown in the top. Statistics: non-parametric ANOVA; **p < 0.01.