Figure 3.

Maternal ethanol administration (2 g/kg/d, E10–E15) compared with isocaloric control and untreated control groups affects CCR2⁺ and MCH⁺ neurons in the LH of adolescent offspring (n = 7/group/sex), as measured using IF. A, Ethanol compared with control groups significantly increased the density of CCR2⁺ and MCH⁺ single-labeled neurons in both female and male offspring, with the effects in females significantly larger. B, This effect in female offspring is illustrated in representative immunostaining images of CCR2⁺ and MCH⁺ single-labeled neurons. C, Ethanol increased the density of CCR2⁺/MCH⁺ double-labeled neurons in both female and male offspring, again with this effect significantly stronger in females. D, This effect of ethanol on double-labeled neurons is illustrated in the confocal images from female offspring showing CCR2⁺ (red) and MCH⁺ (green) single-labeled neurons and CCR2⁺/MCH⁺ (yellow) double-labeled neurons in the LH. Top, A few CCR2⁺/MCH⁺ neurons (arrows) in control offspring and many in ethanol-exposed offspring, with those in the white square illustrated (bottom) at higher magnification for CCR2⁺ and MCH⁺ single-labeled and CCR2⁺/MCH⁺ double-labeled neurons. Scale bars, 200 μm. Data are mean ± SEM. *p < 0.05 versus control group. ^#p < 0.05 versus male groups.