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. 2023 Feb 8;12(4):546.
doi: 10.3390/cells12040546.

Choline Supplementation Alters Hippocampal Cytokine Levels in Adolescence and Adulthood in an Animal Model of Fetal Alcohol Spectrum Disorders

Jessica A Baker  1 , Tamara S Bodnar  2 , Kristen R Breit  1   3 , Joanne Weinberg  2 , Jennifer D Thomas  1
Affiliations

Choline Supplementation Alters Hippocampal Cytokine Levels in Adolescence and Adulthood in an Animal Model of Fetal Alcohol Spectrum Disorders

Jessica A Baker et al. Cells. .

Abstract

Alcohol (ethanol) exposure during pregnancy can adversely affect development, with long-lasting consequences that include neuroimmune, cognitive, and behavioral dysfunction. Alcohol-induced alterations in cytokine levels in the hippocampus may contribute to abnormal cognitive and behavioral outcomes in individuals with fetal alcohol spectrum disorders (FASD). Nutritional intervention with the essential nutrient choline can improve hippocampal-dependent behavioral impairments and may also influence neuroimmune function. Thus, we examined the effects of choline supplementation on hippocampal cytokine levels in adolescent and adult rats exposed to alcohol early in development. From postnatal day (PD) 4-9 (third trimester-equivalent), Sprague-Dawley rat pups received ethanol (5.25 g/kg/day) or sham intubations and were treated with choline chloride (100 mg/kg/day) or saline from PD 10-30; hippocampi were collected at PD 35 or PD 60. Age-specific ethanol-induced increases in interferon gamma (IFN-γ), tumor necrosis factor alpha (TNF-α), and keratinocyte chemoattractant/human growth-regulated oncogene (KC/GRO) were identified in adulthood, but not adolescence, whereas persistent ethanol-induced increases of interleukin-6 (IL-6) levels were present at both ages. Interestingly, choline supplementation reduced age-related changes in interleukin-1 beta (IL-1β) and interleukin-5 (IL-5) as well as mitigating the long-lasting increase in IFN-γ in ethanol-exposed adults. Moreover, choline influenced inflammatory tone by modulating ratios of pro- to -anti-inflammatory cytokines. These results suggest that ethanol-induced changes in hippocampal cytokine levels are more evident during adulthood than adolescence, and that choline can mitigate some effects of ethanol exposure on long-lasting inflammatory tone.

Keywords: FASD; alcohol; choline; cytokine; ethanol; hippocampus; neuroimmune.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Timeline of experimental procedures. GD: gestational day, PD: postnatal day.
Figure 2
Figure 2
Hippocampal cytokine profiles during adolescence and adulthood. Overall, analysis demonstrates a noticeable clustering of cytokines patterns by age. Rows represent groups, as indicated, and the columns represent mean cytokine levels (z-scored data) for each group. Colors demonstrate deviations from the mean of zero, as indicated in the color key. EtOH: Ethanol.
Figure 3
Figure 3
Dendrogram of treatment groups during adolescence or adulthood based on overall hippocampal cytokine profiles. In adolescence, choline supplemented groups clustered together as did non-supplemented groups, regardless of ethanol exposure. In contrast, ethanol-exposed subjects differed the most from all other groups in adulthood. EtOH: Ethanol. Created using BioRender.com.
Figure 4
Figure 4
Dendrograms of cytokine grouping during adolescence and adulthood. Cytokine clustering shows similarities in adolescence and adulthood, despite differential cluster of groups within each age. Created using BioRender.com.
Figure 5
Figure 5
Developmental alcohol exposure and choline supplementation altered levels at cytokines during adolescence and adulthood. The following hippocampal cytokines at adolescence and adulthood are depicted in the graphs above: (A) IL-6 (EtOH > Sham, * p < 0.01; Adolescent > Adult, p < 0.01), (B) IFN-γ (EtOH + Choline < EtOH + Saline, * p < 0.05; Adolescent > Adult, p < 0.001), (C) KC/GRO (Adult: EtOH > Saline, * p < 0.05), (D) TNF-α (Adult: EtOH > Saline, * p < 0.05), (E) IL-5 (Adolescent: Choline < Saline, ^ p = 0.07; Adolescent > Adult, p < 0.001), (F) IL-13 (Choline < Saline, p = 0.12; Adolescent < Adult, p < 0.05), and (G) IL-1β (Adolescent > Adult, p < 0.001). n = 18–24 per group.
Figure 6
Figure 6
Change in hippocampal cytokine profiles across ages in each treatment group. Heatmaps depicting overall cytokine profiles the difference scores of adult cytokine levels compared to adolescent cytokines levels. Rows represent groups, as indicated, and the columns represent mean cytokine levels (z-scored data) for each group. Colors demonstrate deviations from the mean of zero, as indicated in the color key. EtOH: Ethanol.
Figure 7
Figure 7
Clustering of cytokines across development in each treatment group. Differential clustering of cytokine changes across development are depicted in the treatment groups: (A) Sham + Saline, (B) Sham + Choline, (C) EtOH + Saline, (D) EtOH + Choline. The mean difference score for each cytokine within each group is depicted with each cytokine symbol. Negative numbers represent decrease in cytokine levels from adolescent to adulthood, whereas positive numbers show an increase. EtOH: Ethanol. Created using BioRender.com.
Figure 8
Figure 8
Developmental EtOH exposure and choline supplementation alter cytokine levels across development. Difference scores from adolescent to adulthood are used to show change in cytokine levels across development for (A) KCGRO (EtOH > Sham, * p < 0.05), (B) TNF-α (EtOH > Sham, * p < 0.01), (C) IL-1β (Choline < Saline, * p < 0.001), (D) IL-5 (Choline < Saline, * p < 0.001), (E) IFN-γ (Sham + Saline < EtOH + Saline & Sham + Choline * p’s < 0.05), and (F) IL-13 (Sham + Saline < all other groups, * p’s < 0.01). n = 19–24 per group.
Figure 9
Figure 9
EtOH exposure and choline supplementation altered ratio of pro-inflammatory to anti-inflammatory cytokines. (A) Ratio of pro-inflammatory cytokine, TNF-α, to anti-inflammatory cytokine, IL-10 during adolescence and adulthood. EtOH exposure and choline supplementation altered the ratios of TNF-α:IL-10 in adults (EtOH + Saline > Sham + Saline, * p < 0.05), but not adolescents. (B) Ratio of pro-inflammatory cytokine, IFN-γ, to anti-inflammatory cytokine, IL-5 during adolescence and adulthood. Choline supplementation altered ratios of IFN-γ:IL-5 in adults (Choline < Saline, * p < 0.05) but not adolescents. n = 18–19 per group.
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References

    1. Denny C.H., Acero C.S., Naimi T.S., Kim S.Y. Consumption of Alcohol Beverages and Binge Drinking among Pregnant Women Aged 18-44 Years—United States, 2015–2017. MMWR Morb. Mortal Wkly. Rep. 2019;68:365–368. doi: 10.15585/mmwr.mm6816a1. - DOI - PMC - PubMed
    1. Denny C.H., Tsai J., Floyd R.L., Green P. Alcohol use among pregnant and nonpregnant women of childbearing age—United States, 1991-2005. MMWR Morb. Mortal Wkly. Rep. 2009;58:529–532. - PubMed
    1. Riley E.P., Infante M.A., Warren K.R. Fetal alcohol spectrum disorders: An overview. Neuropsychol. Rev. 2011;21:73–80. doi: 10.1007/s11065-011-9166-x. - DOI - PMC - PubMed
    1. May P.A., Hasken J.M., Hooper S.R., Hedrick D.M., Jackson-Newsom J., Mullis C.E., Dobyns E., Kalberg W.O., Buckley D., Robinson L.K., et al. Estimating the community prevalence, child traits, and maternal risk factors of fetal alcohol spectrum disorders (FASD) from a random sample of school children. Drug Alcohol Depend. 2021;227:108918. doi: 10.1016/j.drugalcdep.2021.108918. - DOI - PMC - PubMed
    1. May P.A., Chambers C.D., Kalberg W.O., Zellner J., Feldman H., Buckley D., Kopald D., Hasken J.M., Xu R., Honerkamp-Smith G., et al. Prevalence of Fetal Alcohol Spectrum Disorders in 4 US Communities. JAMA. 2018;319:474–482. doi: 10.1001/jama.2017.21896. - DOI - PMC - PubMed

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