Abstinence From Moderate Alcohol Self-Administration Alters Progenitor Cell Proliferation and Differentiation in Multiple Brain Regions of Male and Female P Rats

Authors

Jun He, David H. Overstreet, Fulton T. Crews

Abstract

Acute and chronic ethanol exposure has been found to decrease hippocampal neurogenesis, reduce dendritic differentiation of new neurons, and increase cell death. Interestingly, abstinence from such treatment increases hippocampal neurogenesis and microglial genesis across several brain regions. The goal of the current investigation was to study cellular alterations on neuro- and cell-genesis during abstinence following alcohol self-administration using alcohol-preferring rats (P rats). Male and female P rats were given the choice of drinking 10% alcohol in water or pure water for 7 weeks. Social interaction behavioral assessments were conducted at 5 hours upon removal of alcohol, followed by bromo-deoxyuridine (BrdU, 150 mg/kg × 1/d × 14 d) injections to label proliferating cells. Animals were then killed 4 weeks later to conduct immunohistochemical and confocal analyses using antibodies against BrdU and other phenotypic markers (NeuN for mature neurons; Iba-1 for microglia; GFAP for astrocytes; and NG2 for oligodendrocyte progenitors). Mild alcohol withdrawal anxiety was detected by reduction in social interactions. The number of hippocampal BrdU+ cells was increased approximately 50% during alcohol abstinence (26 ± 2.8 in controls vs. 39 ± 4 in alcohol group). BrdU+ cells were also increased in the substantia nigra (SN) approximately 65% in the alcohol abstinent group (12 ± 1 in controls vs. 19 ± 1.5 in alcohol group). No gender differences were found. Confocal analyses indicated that approximately 75% of co-localization of BrdU+ cells with NeuN in the hippocampal dentate gyrus (DG) resulting a net increase in neurogenesis in the alcohol abstinent group compared to controls. In the cingulum, a greater proportion of BrdU+ cells were co-localized with NG2 in the alcohol abstinent group indicating increased differentiation toward oligodendrocyte progenitors in both genders. However, the phenotype of the BrdU+ cells in SN and other brain regions were not identified by NeuN, Iba-1, GFAP, or NG2 suggesting that these BrdU+ cells probably remain in a nondifferentiated stage. These data indicate that abstinence from moderate alcohol drinking increases hippocampal neurogenesis, cingulate NG2 differentiation, and SN undifferentiated cell proliferation in both males and females. Such cellular alteration during abstinence could contribute to the spontaneous partial restoration of cognitive deficits upon sobriety.