Percentage of GFAP/BrdU double-labeled cells in the dentate gyrus.

<p>There was no effect of MD on the percentage of double labeled cells in both males and females (F_(3,24) = 0.40; p = 0.54). No effect of sex was found (F_(1,24) = 0.56; p = 0.46).</p

Dentate gyrus cell proliferation (Ki-67) on PND21.

<p>A. In males, MD affected dentate cell proliferation rate at PND21 (F_(3,28) = 3,2, p = 0.043). Post-hoc analysis revealed a decrease in MD-glucose injected animals (p<0.05). B. In females, there was no effect of treatment (F_(3,28) = 0.41 p = 0.75), although a significant lower number of Ki-67 positive cells was found in females when compared to males (F_(1,56) = 8.56; p = 0.005).</p

Licking and grooming on PND1–7 in control and MD litters.

<p>A significant increase in LG was found on PND4 both in males and females compared to their same-sex controls. (PND4 LG-scores: males F_(3,21) = 7.31; p = 0.002; post-hoc: MD males>CON males = MD females>CON females, p<0.05).</p

Number of newborn surviving cells (BrdU+) in the dentate gyrus on PND21.

<p>There was a significant effect of sex, but not of treatment on BrdU positive cell numbers. A. Maternal deprivation did not alter BrdU+ cell numbers in males (F_(3,28) = 0.40, p = 0.75), or (B) females (F_(3,28) = 1.1, p = 0.37). Irrespective of MD, females had an overall lower number of BrdU-positive cells (F_(1,51) = 29.8; p<0.0001).</p

Arch back nursing.

<p>Arch back nursing towards the entire litter in control and MD litters on PND1–7. There is a significant increase after PND3 (F = 7.89, p = 0.02).</p

Doublecortin (DCX) -positive neuron numbers on PND21.

<p>A significant treatment×sex interaction revealed a differential effect of MD on males versus females (F_(3,48) = 8.04; p<0.0001). A. An increase in DCX+ cell number was found in deprived males (F_(3,28) = 4.3, p = 0.018; post-hoc: CONU = CONS(3,28) = 4.65, p = 0.013; post-hoc: CONU = CONS>MDS = MDG, at least p<0.05) when compared to controls. A significant effect of sex indicates a general lower amount of DCX+ cells in females (F_(1,48) = 65.80; p<0.0001).</p

Doublecortin (DCX) -positive neurons.

<p>A. Photo of the dentate gyrus of a 21 day old CONS male showing extensive immunostaining of DCX in the subgranular zone (sgz) and the first third of the granular cell layer (GCL) with dendrites extending through the granular cell layer (GCL) into the molecular layer (ML). B. High power photomicrograph showing details of the DCX+ cell bodies located in the SGZ and GCL, with extending dendrites in the GCL.</p

Granular cell layer (GCL) volume on PND21.

<p>There was no effect of any of the treatments on granular cell layer volume. However, significantly lower GCL volumes were found in females (F_(1.51) = 122.4; p<0.0001).</p

Astrocyte numbers.

<p>GFAP-positive astrocyte numbers determined stereologically in the entire hippocampal dentate gyrus on PND21. A. Maternal deprivation did not affect GFAP+ cell number in males (F_(3,28) = 1.3, p = 0.28), B. nor females (F_(3,28) = 0.05, p = 0.99). However, a significant effect of sex on GFAP+ cell number was found (F_(3,56) = 25.01; p<0.0001).</p

GFAP/BrdU double labeling.

<p>Immunohistochemical double labeling for GFAP and BrdU shows single GFAP+ astrocytes in the hilus with their processes occasionally extending into the sgz. GFAP+ cells reveal brown DAB-staining in their processes and cytoplasm whereas the nucleus is devoid of staining. In the GCL, BrdU+ single cells are stained black by DAB-nickel as indicated (BrdU+) in the granular cell layer (GCL). Shown on the left is a BrdU/GFAP double labeled cell (arrow).</p