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Journal of Neurodevelopmental Disorders
Published in: Journal of Neurodevelopmental Disorders 1/2022

Open Access 01-12-2022 | Fetal Alcohol Spectrum Disorder | Research

mRNA expression analysis of the hippocampus in a vervet monkey model of fetal alcohol spectrum disorder

Authors: Rob F. Gillis, Roberta M. Palmour

Published in: Journal of Neurodevelopmental Disorders | Issue 1/2022

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Abstract

Background

Fetal alcohol spectrum disorders (FASD) are common, yet preventable developmental disorders that stem from prenatal exposure to alcohol. This exposure leads to a wide array of behavioural and physical problems with a complex and poorly defined biological basis.
Molecular investigations to date predominantly use rodent animal models, but because of genetic, developmental and social behavioral similarity, primate models are more relevant. We previously reported reduced cortical and hippocampal neuron levels in an Old World monkey (Chlorocebus sabaeus) model with ethanol exposure targeted to the period of rapid synaptogenesis and report here an initial molecular study of this model. The goal of this study was to evaluate mRNA expression of the hippocampus at two different behavioural stages (5 months, 2 years) corresponding to human infancy and early childhood.

Methods

Offspring of alcohol-preferring or control dams drank a maximum of 3.5 g ethanol per kg body weight or calorically matched sucrose solution 4 days per week during the last 2 months of gestation. Total mRNA expression was measured with the Affymetrix GeneChip Rhesus Macaque Genome Array in a 2 × 2 study design that interrogated two independent variables, age at sacrifice, and alcohol consumption during gestation.

Results and discussion

Statistical analysis identified a preferential downregulation of expression when interrogating the factor ‘alcohol’ with a balanced effect of upregulation vs. downregulation for the independent variable ‘age’. Functional exploration of both independent variables shows that the alcohol consumption factor generates broad functional annotation clusters that likely implicate a role for epigenetics in the observed differential expression, while the variable age reliably produced functional annotation clusters predominantly related to development. Furthermore, our data reveals a novel connection between EFNB1 and the FASDs; this is highly plausible both due to the role of EFNB1 in neuronal development as well as its central role in craniofrontal nasal syndrome (CFNS). Fold changes for key genes were subsequently confirmed via qRT-PCR.

Conclusion

Prenatal alcohol exposure leads to global downregulation in mRNA expression. The cellular interference model of EFNB1 provides a potential clue regarding how genetically susceptible individuals may develop the phenotypic triad generally associated with classic fetal alcohol syndrome.
Appendix
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Literature
1.
Jones KL, Smith DW, Ulleland CN, Streissguth P. Pattern of malformation in offspring of chronic alcoholic mothers. Lancet. 1973;1(7815):1267–71.PubMedCrossRef
2.
Lemoine P, Harousseau, H., Borteyru, J.P., Menuet, J.C., 1968. Les enfants des parents alcoholiques; anomalies observees a propos de 127 cas. Quest Medical. 1968;25:476-82.
3.
Goulden KJ. Are FASD guidelines practical and sustainable? CMAJ. 2005;173(9):1070; author reply -1.
4.
Viljoen DL, Gossage JP, Brooke L, Adnams CM, Jones KL, Robinson LK, et al. Fetal alcohol syndrome epidemiology in a South African community: a second study of a very high prevalence area. J Stud Alcohol. 2005;66(5):593–604.PubMedPubMedCentralCrossRef
5.
Streissguth AP, Dehaene P. Fetal alcohol syndrome in twins of alcoholic mothers: concordance of diagnosis and IQ. Am J Med Genet. 1993;47(6):857–61.PubMedCrossRef
6.
Warren KR, Li TK. Genetic polymorphisms: impact on the risk of fetal alcohol spectrum disorders. Birth Defects Res A Clin Mol Teratol. 2005;73(4):195–203.PubMedCrossRef
7.
Goodlett CR, Gilliam DM, Nichols JM, West JR. Genetic influences on brain growth restriction induced by development exposure to alcohol. Neurotoxicology. 1989;10(3):321–34.PubMed
8.
Gilliam DM, Kotch LE. Dose-related growth deficits in LS but not SS mice prenatally exposed to alcohol. Alcohol. 1996;13(1):47–51.PubMedCrossRef
9.
Hard ML, Abdolell M, Robinson BH, Koren G. Gene-expression analysis after alcohol exposure in the developing mouse. J Lab Clin Med. 2005;145(1):47–54.PubMedCrossRef
10.
Zhang C, Frazier JM, Chen H, Liu Y, Lee JA, Cole GJ. Molecular and morphological changes in zebrafish following transient ethanol exposure during defined developmental stages. Neurotoxicol Teratol. 2014;44:70–80.PubMedPubMedCentralCrossRef
11.
El Shawa H, Abbott CW 3rd, Huffman KJ. Prenatal ethanol exposure disrupts intraneocortical circuitry, cortical gene expression, and behavior in a mouse model of FASD. J Neurosci. 2013;33(48):18893–905.PubMedPubMedCentralCrossRef
12.
Stringer RL, Laufer BI, Kleiber ML, Singh SM. Reduced expression of brain cannabinoid receptor 1 (Cnr1) is coupled with an increased complementary micro-RNA (miR-26b) in a mouse model of fetal alcohol spectrum disorders. Clin Epigenetics. 2013;5(1):14.PubMedPubMedCentralCrossRef
13.
Kleiber ML, Laufer BI, Wright E, Diehl EJ, Singh SM. Long-term alterations to the brain transcriptome in a maternal voluntary consumption model of fetal alcohol spectrum disorders. Brain research. 2012;1458:18–33.PubMedCrossRef
14.
Downing C, Flink S, Florez-McClure ML, Johnson TE, Tabakoff B, Kechris KJ. Gene expression changes in C57BL/6J and DBA/2J mice following prenatal alcohol exposure. Alcohol Clin Exp Res. 2012;36(9):1519–29.PubMedPubMedCentralCrossRef
15.
Green ML, Singh AV, Zhang Y, Nemeth KA, Sulik KK, Knudsen TB. Reprogramming of genetic networks during initiation of the Fetal Alcohol Syndrome. Dev Dyn. 2007;236(2):613–31.PubMedCrossRef
16.
Hashimoto-Torii K, Kawasawa YI, Kuhn A, Rakic P. Combined transcriptome analysis of fetal human and mouse cerebral cortex exposed to alcohol. Proc Natl Acad Sci U S A. 2011;108(10):4212–7.PubMedPubMedCentralCrossRef
17.
Kimura KA, Chiu J, Reynolds JN, Brien JF. Effect of chronic prenatal ethanol exposure on nitric oxide synthase I and III proteins in the hippocampus of the near-term fetal guinea pig. Neurotoxicol Teratol. 1999;21(3):251–9.PubMedCrossRef
18.
Mandal C, Park KS, Jung KH, Chai YG. Ethanol-related alterations in gene expression patterns in the developing murine hippocampus. Acta Biochim Biophys Sin (Shanghai). 2015;47(8):581–7.PubMedCrossRef
19.
Lunde-Young R, Ramirez J, Naik V, Orzabal M, Lee J, Konganti K, et al. Hippocampal transcriptome reveals novel targets of FASD pathogenesis. Brain Behav. 2019;9(7):e01334.PubMedPubMedCentralCrossRef
20.
Alberry BLJ, Castellani CA, Singh SM. Hippocampal transcriptome analysis following maternal separation implicates altered RNA processing in a mouse model of fetal alcohol spectrum disorder. J Neurodev Disord. 2020;12(1):15.PubMedPubMedCentralCrossRef
21.
Ehrhart F, Roozen S, Verbeek J, Koek G, Kok G, van Kranen H, et al. Review and gap analysis: molecular pathways leading to fetal alcohol spectrum disorders. Mol Psychiatry. 2019;24(1):10–7.PubMedCrossRef
22.
Howard RJ, Slesinger PA, Davies DL, Das J, Trudell JR, Harris RA. Alcohol-binding sites in distinct brain proteins: the quest for atomic level resolution. Alcohol Clin Exp Res. 2011;35(9):1561–73.PubMedPubMedCentral
23.
Freund G. Neurobiological relationships between aging and alcohol abuse. Recent Dev Alcohol. 1984;2:203–21.PubMedCrossRef
24.
Korbo L. Glial cell loss in the hippocampus of alcoholics. Alcohol Clin Exp Res. 1999;23(1):164–8.PubMedCrossRef
25.
White AM, Swartzwelder HS. Hippocampal function during adolescence: a unique target of ethanol effects. Annals of the New York Academy of Sciences. 2004;1021:206–20.PubMedCrossRef
26.
Medina KL, Schweinsburg AD, Cohen-Zion M, Nagel BJ, Tapert SF. Effects of alcohol and combined marijuana and alcohol use during adolescence on hippocampal volume and asymmetry. Neurotoxicol Teratol. 2007;29(1):141–52.PubMedCrossRef
27.
Nagel BJ, Schweinsburg AD, Phan V, Tapert SF. Reduced hippocampal volume among adolescents with alcohol use disorders without psychiatric comorbidity. Psychiatry research. 2005;139(3):181–90.PubMedPubMedCentralCrossRef
28.
Morris SA, Eaves DW, Smith AR, Nixon K. Alcohol inhibition of neurogenesis: a mechanism of hippocampal neurodegeneration in an adolescent alcohol abuse model. Hippocampus. 2010;20(5):596–607.PubMedPubMedCentral
29.
Livy DJ, Miller EK, Maier SE, West JR. Fetal alcohol exposure and temporal vulnerability: effects of binge-like alcohol exposure on the developing rat hippocampus. Neurotoxicol Teratol. 2003;25(4):447–58.PubMedCrossRef
30.
Barnes DE, Walker DW. Prenatal ethanol exposure permanently reduces the number of pyramidal neurons in rat hippocampus. Brain research. 1981;227(3):333–40.PubMedCrossRef
31.
McGoey TN, Reynolds JN, Brien JF. Chronic prenatal ethanol exposure-induced decrease of guinea pig hippocampal CA1 pyramidal cell and cerebellar Purkinje cell density. Can J Physiol Pharmacol. 2003;81(5):476–84.PubMedCrossRef
32.
Burke MW, Ptito M, Ervin FR, Palmour RM. Hippocampal neuron populations are reduced in vervet monkeys with fetal alcohol exposure. Dev Psychobiol. 2015;57(4):470–85.PubMedPubMedCentralCrossRef
33.
Lee AG, Hagenauer M, Absher D, Morrison KE, Bale TL, Myers RM, et al. Stress amplifies sex differences in primate prefrontal profiles of gene expression. Biol Sex Differ. 2017;8(1):36.PubMedPubMedCentralCrossRef
34.
Palmour RM, Mulligan J, Howbert JJ, Ervin F. Of monkeys and men: vervets and the genetics of human-like behaviors. Am J Human Genetics. 1997;61(3):481–8.CrossRef
35.
Labonte B, Engmann O, Purushothaman I, Menard C, Wang J, Tan C, et al. Sex-specific transcriptional signatures in human depression. Nat Med. 2017;23(9):1102–11.PubMedPubMedCentralCrossRef
36.
Ervin FR, Palmour RM, Young SN, Guzman-Flores C, Juarez J. Voluntary consumption of beverage alcohol by vervet monkeys: population screening, descriptive behavior and biochemical measures. Pharmacol Biochem Behav. 1990;36(2):367–73.PubMedCrossRef
37.
Palmour RM, Ervin FR, Baker GB, Young SN. An amino acid mixture deficient in phenylalanine and tyrosine reduces cerebrospinal fluid catecholamine metabolites and alcohol consumption in vervet monkeys. Psychopharmacology (Berl). 1998;136(1):1–7.PubMedCrossRef
38.
Association AVM. AVMA Guidelines for the Euthanasia of Animals. Schaumber, Il2013.
39.
Gentleman RC, Carey VJ, Bates DM, Bolstad B, Dettling M, Dudoit S, et al. Bioconductor: open software development for computational biology and bioinformatics. Genome Biol. 2004;5(10):R80.PubMedPubMedCentralCrossRef
40.
Irizarry RA, Hobbs B, Collin F, Beazer-Barclay YD, Antonellis KJ, Scherf U, et al. Exploration, normalization, and summaries of high density oligonucleotide array probe level data. Biostatistics. 2003;4(2):249–64.PubMedCrossRef
41.
Blazejczyk M, Nadon. FlexArray: A statistical data analysis software for gene expression microarrays. 2007.
42.
Liu WM, Mei R, Di X, Ryder TB, Hubbell E, Dee S, et al. Analysis of high density expression microarrays with signed-rank call algorithms. Bioinformatics. 2002;18(12):1593–9.PubMedCrossRef
43.
44.
Miron M, Nadon R. Inferential literacy for experimental high-throughput biology. Trends Genet. 2006;22(2):84–9.PubMedCrossRef
45.
Storey JD. A direct approach to false discovery rates. J Roy Stat Soc B. 2002;64:479–98.CrossRef
46.
Chen X, Robinson DG, Storey JD. The functional false discovery rate with applications to genomics. Biostatistics. 2021;22(1):68–81.PubMedCrossRef
47.
Rogic S, Wong A, Pavlidis P. Meta-analysis of gene expression patterns in animal models of prenatal alcohol exposure suggests role for protein synthesis inhibition and chromatin remodeling. Alcohol Clin Exp Res. 2016;40(4):717–27.PubMedPubMedCentralCrossRef
48.
Garro AJ, McBeth DL, Lima V, Lieber CS. Ethanol consumption inhibits fetal DNA methylation in mice: implications for the fetal alcohol syndrome. Alcohol Clin Exp Res. 1991;15(3):395–8.PubMedCrossRef
49.
Laufer BI, Mantha K, Kleiber ML, Diehl EJ, Addison SM, Singh SM. Long-lasting alterations to DNA methylation and ncRNAs could underlie the effects of fetal alcohol exposure in mice. Disease Models Mechanisms. 2013;6(4):977–92.PubMedPubMedCentral
50.
Liu Y, Balaraman Y, Wang G, Nephew KP, Zhou FC. Alcohol exposure alters DNA methylation profiles in mouse embryos at early neurulation. Epigenetics. 2009;4(7):500–11.PubMedCrossRef
51.
Portales-Casamar E, Lussier AA, Jones MJ, MacIsaac JL, Edgar RD, Mah SM, et al. DNA methylation signature of human fetal alcohol spectrum disorder. Epigenetics Chromatin. 2016;9:25.PubMedPubMedCentralCrossRef
52.
Balaraman S, Tingling JD, Tsai PC, Miranda RC. Dysregulation of microRNA expression and function contributes to the etiology of fetal alcohol spectrum disorders. Alcohol Res: Current Reviews. 2013;35(1):18–24.
53.
Morey JS, Ryan JC, Van Dolah FM. Microarray validation: factors influencing correlation between oligonucleotide microarrays and real-time PCR. Biological Procedures Online. 2006;8:175–93.PubMedPubMedCentralCrossRef
54.
55.
Etienne W, Meyer MH, Peppers J, Meyer RA Jr. Comparison of mRNA gene expression by RT-PCR and DNA microarray. Biotechniques. 2004;36(4):618 -20, 22, 24-6.PubMedCrossRef
56.
Rajeevan MS, Vernon SD, Taysavang N, Unger ER. Validation of array-based gene expression profiles by real-time (kinetic) RT-PCR. J Molecular Diagnostics : JMD. 2001;3(1):26–31.CrossRef
57.
Bruckner K, Pasquale EB, Klein R. Tyrosine phosphorylation of transmembrane ligands for Eph receptors. Science. 1997;275(5306):1640–3.PubMedCrossRef
58.
Wilkinson DG. Multiple roles of EPH receptors and ephrins in neural development. Nature Reviews Neurosci. 2001;2(3):155–64.CrossRef
59.
Palmer A, Zimmer M, Erdmann KS, Eulenburg V, Porthin A, Heumann R, et al. EphrinB phosphorylation and reverse signaling: regulation by Src kinases and PTP-BL phosphatase. Molecular cell. 2002;9(4):725–37.PubMedCrossRef
60.
Beckmann MP, Cerretti DP, Baum P, Vanden Bos T, James L, Farrah T, et al. Molecular characterization of a family of ligands for eph-related tyrosine kinase receptors. EMBO J. 1994;13(16):3757–62.PubMedPubMedCentralCrossRef
61.
Davis S, Gale NW, Aldrich TH, Maisonpierre PC, Lhotak V, Pawson T, et al. Ligands for EPH-related receptor tyrosine kinases that require membrane attachment or clustering for activity. Science. 1994;266(5186):816–9.PubMedCrossRef
62.
Klein R. Eph/ephrin signaling in morphogenesis, neural development and plasticity. Current Opinion Cell Biol. 2004;16(5):580–9.PubMedCrossRef
63.
Bush JO, Soriano P. Ephrin-B1 regulates axon guidance by reverse signaling through a PDZ-dependent mechanism. Genes Development. 2009;23(13):1586–99.PubMedPubMedCentralCrossRef
64.
Nguyen AQ, Sutley S, Koeppen J, Mina K, Woodruff S, Hanna S, et al. Astrocytic Ephrin-B1 Controls Excitatory-Inhibitory Balance in Developing Hippocampus. J Neurosci. 2020;40(36):6854–71.PubMedPubMedCentralCrossRef
65.
Koeppen J, Nguyen AQ, Nikolakopoulou AM, Garcia M, Hanna S, Woodruff S, et al. Functional Consequences of Synapse Remodeling Following Astrocyte-Specific Regulation of Ephrin-B1 in the Adult Hippocampus. J Neuroscience. 2018;38(25):5710–26.CrossRef
66.
Wieland I, Jakubiczka S, Muschke P, Cohen M, Thiele H, Gerlach KL, et al. Mutations of the ephrin-B1 gene cause craniofrontonasal syndrome. Am J Human Genetics. 2004;74(6):1209–15.CrossRef
67.
Wieland I, Weidner C, Ciccone R, Lapi E, McDonald-McGinn D, Kress W, et al. Contiguous gene deletions involving EFNB1, OPHN1, PJA1 and EDA in patients with craniofrontonasal syndrome. Clinical Genetics. 2007;72(6):506–16.PubMedCrossRef
68.
Wallis D, Lacbawan F, Jain M, Der Kaloustian VM, Steiner CE, Moeschler JB, et al. Additional EFNB1 mutations in craniofrontonasal syndrome. Am J Med Genetics Part A. 2008;146A(15):2008–12.CrossRef
69.
Twigg SR, Kan R, Babbs C, Bochukova EG, Robertson SP, Wall SA, et al. Mutations of ephrin-B1 (EFNB1), a marker of tissue boundary formation, cause craniofrontonasal syndrome. Proc Natl Acad Sci U S A. 2004;101(23):8652–7.PubMedPubMedCentralCrossRef
70.
Kapusta L, Brunner HG, Hamel BC. Craniofrontonasal dysplasia. European journal of pediatrics. 1992;151(11):837–41.PubMedCrossRef
71.
Goyal M, Pradhan G, Wieland I, Kapoor S. Craniofrontonasal syndrome: atrial septal defect with a novel EFNB1 gene mutation. The Cleft Palate-Craniofacial J. 2015;52(2):234–6.CrossRef
72.
Wieland I, Makarov R, Reardon W, Tinschert S, Goldenberg A, Thierry P, et al. Dissecting the molecular mechanisms in craniofrontonasal syndrome: differential mRNA expression of mutant EFNB1 and the cellular mosaic. Eur J Human Genet. 2008;16(2):184–91.CrossRef
73.
Wieacker P, Wieland I. Clinical and genetic aspects of craniofrontonasal syndrome: towards resolving a genetic paradox. Mol Genet Metab. 2005;86(1-2):110–6.PubMedCrossRef
74.
Niethamer TK, Larson AR, O'Neill AK, Bershteyn M, Hsiao EC, Klein OD, et al. EPHRIN-B1 Mosaicism Drives Cell Segregation in Craniofrontonasal Syndrome hiPSC-Derived Neuroepithelial Cells. Stem Cell Reports. 2017;8(3):529–37.PubMedPubMedCentralCrossRef
75.
Twigg SR, Babbs C, van den Elzen ME, Goriely A, Taylor S, McGowan SJ, et al. Cellular interference in craniofrontonasal syndrome: males mosaic for mutations in the X-linked EFNB1 gene are more severely affected than true hemizygotes. Hum Mol Genet. 2013;22(8):1654–62.PubMedPubMedCentralCrossRef
76.
Babbs C, Stewart HS, Williams LJ, Connell L, Goriely A, Twigg SR, et al. Duplication of the EFNB1 gene in familial hypertelorism: imbalance in ephrin-B1 expression and abnormal phenotypes in humans and mice. Human Mutation. 2011;32(8):930–8.PubMedPubMedCentralCrossRef
77.
Maier SE, West JR. Regional differences in cell loss associated with binge-like alcohol exposure during the first two trimesters equivalent in the rat. Alcohol. 2001;23(1):49–57.PubMedCrossRef
78.
Idanpaan-Heikkila J, Jouppila P, Akerblom HK, Isoaho R, Kauppila E, Koivisto M. Elimination and metabolic effects of ethanol in mother, fetus, and newborn infant. Am J Obstet Gynecol. 1972;112(3):387–93.PubMedCrossRef
79.
Burke MW, Palmour RM, Ervin FR, Ptito M. Neuronal reduction in frontal cortex of primates after prenatal alcohol exposure. Neuroreport. 2009;20(1):13–7.PubMedCrossRef
Metadata
Title
mRNA expression analysis of the hippocampus in a vervet monkey model of fetal alcohol spectrum disorder
Authors
Rob F. Gillis
Roberta M. Palmour
Publication date
01-12-2022
Publisher
BioMed Central
Published in
Journal of Neurodevelopmental Disorders / Issue 1/2022
Print ISSN: 1866-1947
Electronic ISSN: 1866-1955
DOI
https://doi.org/10.1186/s11689-022-09427-z

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