A. Number of NPY positive (NPY+) cells and percentage of NPY+ cells referred to the total number of cells in the arcuate nucleus (ARC) in the hypothalamus of 25-day-old male and female offspring of dams with free access to standard chow diet (controls), the offspring of 20% calorie restricted dams during the first 12 days of pregnancy (CR), and CR rats daily supplemented with physiological doses of leptin throughout lactation (CR-Leptin). Number of NPY+ cells in the ARC was counted using computerized image analysis software, following immunostaining. Data are mean ± S.E.M. (n = 6-8, coming from at least six different litters). Statistics: in case of interaction within each sex, bars not sharing a common letter (a and b) are significantly different (a≠b) (p < 0.05; LSD post
hoc one-way ANOVA test). B. Representative brain sections immunostained for NPY in the arcuate hypothalamic nucleus of the offspring of 25-day-old male (a) and female (d) offspring of dams with free access to standard chow diet (controls), male (b) and female (e) offspring of 20% calorie restricted dams during the first 12 days of pregnancy (CR), and male (c) and female (f) CR rats daily supplemented with physiological doses of leptin throughout lactation (CR-Leptin). Insets: enlargement of the corresponding framed areas. Abbreviations: ARC, arcuate nucleus; 3v, third ventricle. Scale bar: 100 µm and 50 µm for insets.