Fig 2: (a) Cresyl violet staining shows cortical layers (roman numerals) and underlying white matter (WM) in the normal cerebral hemisphere wall in relation to eight depth zones (a vertical white bar with Arabic numerals). The region delimited for counting of immunopositive neurons was a vertical rectangle of 500 µm width located between the lumen of the lateral ventricle and the surface of cerebral hemisphere at the level of the anterior commissure. Within the region, neurons immunopositive for (b) parvalbumin (PV), (c) calretinin (CR), or (d) neuropeptide Y (NPY) were counted. As indicated with white rectangles, for each of the counting regions, example areas with neurons immunopositive for PV (b1), CR (c1, c2), and NPY (d1, d2) are shown at higher magnification. Each bar shows 100 µm length

Fig 3: Immunohistochemical analysis of NPY in rpWAT depot.Immunoreactive NPY area (NPY+) - expressed as µm/mm2 (left axis) and as µm/adipocyte (right axis) - in retroperitoneal white adipose tissue (rpWAT) of 25 d (A and B) and 6 m (C and D), male (A and C) and female (B and D) offspring of controls (C) and 20% caloric restricted dams during gestation (CR) and their corresponding representative immunostained sections (C on the left and CR on the right). Scale bar: 20 µm and 50 µm for 25 d old and 6 m old animals respectively. Data are mean ± s.e.m. (n = 5–6, coming from at least four different litters). For each animal, 16–20 measurements were performed in different random fields coming from two non consecutive sections of the tissue (7–10 fields per section).* Different from their respective C group (p<0.05; Student's t -test).

Fig 4: mRNA levels of AgRP, CART, NPY, ObRb, POMC and SOCS-3 in hypothalamus in the offspring.Hypothalamic expression levels of AgRP, CART, NPY, ObRb, POMC and SOCS-3 in 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). mRNA levels were measured by qRT-PCR and expressed as a percentage of the value of control male rats. Data are mean ± S.E.M. (n = 10-11, coming from at least six different litters). Statistics: *, CR different from control group (p < 0.05; Student’s t-test); 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).

Fig 5: NPY-immunoreactive profiles in the La at the ultrastructural level. NPY-immunoreactivity is localized in cell bodies frequently displaying indented nuclei (arrow in a), in dendrites (b) synaptically contacted by unlabeled terminals (inset represents magnification of boxed area in b; arrow points at synaptic cleft) and in axon terminals forming symmetric synapses (c arrows point at postsynaptic area in a small unlabeled dendrite). M mitochondrion, uD unlabeled dendrite, uT unlabeled terminal. Scale bar in a is 5,000 nm, in b and c 500 nm