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. Author manuscript; available in PMC: 2023 Nov 1.
Published in final edited form as: Addict Biol. 2022 Nov;27(6):e13238. doi: 10.1111/adb.13238

Figure 4:

Figure 4:

The ectopic Hcrt and MCH neurons seen in newborn rats after ethanol exposure (2 g/kg/day) from embryonic day 10 (E10) to E15 are long lasting, similarly evident with altered morphology in nucleus accumbens core (NAcC) and ventromedial caudate putamen (vmCP) of early-adolescent female rats on postnatal day 30 (P30). (A, B, C) As shown using IF in newborn rats, bar graphs (mean ± SEM, n = 3–4/group) show that Ethanol compared to Control increases in early-adolescent rats the density, size and processes of Hcrt neurons in LH, and compared to these LH neurons, the ectopic Hcrt neurons in NAc and vmCP of Ethanol rats not detected in Control rats are reduced in density, are smaller in size, and have fewer processes. (D) Photomicrographs (40x) of Hcrt neurons (green) stained in brain sections using IF illustrate the increase in density in LH of Ethanol compared to Control rats and the ectopic neurons in the NAcC and vmCP detected in Ethanol but not Control rats. Individual Hcrt neurons in white dotted boxes are further enlarged (upper left corners) to illustrate the ethanol-induced changes in neuron size and number of processes (indicated by white arrowheads). Scale bar, 20 μm. (E, F, G) As shown using IF in newborn rats, bar graphs (mean ± SEM, n = 4–5/group) show that Ethanol compared to Control increases in early-adolescent rats the density, size and number of processes of MCH neurons in LH, and compared to these LH neurons, the ectopic MCH neurons in NAc and vmCP of Ethanol rats not detected in Control rats are reduced in density, are smaller in size, and have fewer processes. (H) (D) Photomicrographs (40x) of MCH neurons (red) stained in brain sections using IF illustrate the increase in density in LH of Ethanol compared to Control rats and the ectopic neurons in the NAcC and vmCP detected in Ethanol but not Control rats. Individual MCH neurons in white dotted boxes are further enlarged (upper left corners) to illustrate the ethanol-induced changes in neuron size and number of processes (indicated by white arrowheads). Scale bar, 20 μm. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Abbreviations: Hcrt: Hypocretin/Orexin; IF: Immunofluorescence; LH: lateral hypothalamus; MCH: Melanin-concentrating hormone; NAcC: Nucleus Accumbens Core; P30: Postnatal day 30; vmCP: ventromedial Caudate Putamen.