Animal Models of Early-Life Adversity

  • Hajar Benmhammed
  • Samer El Hayek
  • Inssaf Berkik
  • Hicham Elmostafi
  • Rim Bousalham
  • Abdelhalem Mesfioui
  • Ali Ouichou
  • Aboubaker El Hessni
Part of the Methods in Molecular Biology book series (MIMB, volume 2011)


From the prenatal period throughout the first years of life, the brain undergoes its most rapid development, a period during which it is highly sensitive to external experiences. The timing of brain development differs from one region to another, as it also differs between substrates, neurotransmitter systems, and central endocrine circuitries. These discontinuities are part of the “critical periods of brain development.” Early-life adversity (ELA), such as exposure to infection, maternal deprivation, and substance use, disrupts the programmed brain development, yielding a myriad of deviations in brain circuitry, stress responsivity, cognitive function, and general health. This is applicable to both humans and animal models.

In our laboratory, several experimental animal designs have been developed that allow investigating the long-lasting consequences of ELA on brain function, cognitive and emotional development, and the risk to develop stress-related psychopathology later in adulthood. This book chapter will provide a review of such animal models, in particular, designs related to infections (LPS-induced), the quality of mother-infant relationship (maternal deprivation and separation), and substance use (ethanol intoxication). The behavior tests, biochemical, and immunohistochemistry assays applied after ELA will be explained. The behavioral tests encompass the open-field, elevated plus maze, forced swim, sucrose preference, Y-maze, object recognition, and Morris water maze tests. These experiments allow the assessment of several outcomes of interest, pertaining to locomotor activity, anxiety-like symptoms, depressive-like symptoms, working memory, recognition memory, spatial memory, and learning performance. The biochemical assays are employed to measure the level of oxidative stress and inflammation in brain areas after application of adversity. Immunohistochemistry puts into perspective the degree of immunoreactivity in the brain subjected to adversity. The findings from our laboratory indicate that the nature and timing of exposure play a critical role in sensitivity to develop neurodevelopmental disorders.

Key words

Early-life adversity Biochemical assays Behavioral tests Immunohistochemistry Neurodevelopmental disorders 


  1. 1.
    Giedd JN, Lalonde FM, Celano MJ, White SL, Wallace GL, Lee NR, Lenroot RK (2009) Anatomical brain magnetic resonance imaging of typically developing children and adolescents. J Am Acad Child Adolesc Psychiatry 48:465–470CrossRefGoogle Scholar
  2. 2.
    Anda RF, Brown DW, Dube SR, Bremner JD, Felitti VJ, Giles WH (2008) Adverse childhood experiences and chronic obstructive pulmonary disease in adults. Am J Prev Med 34:396–403CrossRefGoogle Scholar
  3. 3.
    Dube SR, Fairweather D, Pearson WS, Felitti VJ, Anda RF, Croft JB (2009) Cumulative childhood stress and autoimmune diseases in adults. Psychosom Med 71:243–250CrossRefGoogle Scholar
  4. 4.
    Brown DW, Anda RF, Felitti VJ, Edwards VJ, Malarcher AM, Croft JB, Giles WH (2010) Adverse childhood experiences are associated with the risk of lung cancer: a prospective cohort study. BMC Public Health 10:20CrossRefGoogle Scholar
  5. 5.
    Ball JS, Links PS (2009) Borderline personality disorder and childhood trauma: evidence for a causal relationship. Curr Psychiatry Rep 11:63–68CrossRefGoogle Scholar
  6. 6.
    Bendall S, Jackson HJ, Hulbert CA, McGorry PD (2008) Childhood trauma and psychotic disorders: a systematic, critical review of the evidence. Schizophr Bull 34:568–579CrossRefGoogle Scholar
  7. 7.
    Norman RE, Byambaa M, De R, Butchart A, Scott J, Vos T (2012) The long-term health consequences of child physical abuse, emotional abuse, and neglect: a systematic review and meta-analysis. PLoS Med 9:e1001349CrossRefGoogle Scholar
  8. 8.
    Teicher MH, Samson JA (2013) Childhood maltreatment and psychopathology: a case for ecophenotypic variants as clinically and neurobiologically distinct subtypes. Am J Psychiatry 170:1114–1133CrossRefGoogle Scholar
  9. 9.
    De Bellis MD, Keshavan MS, Spencer S, Hall J (2000) N-Acetylaspartate concentration in the anterior cingulate of maltreated children and adolescents with PTSD. Am J Psychiatry 157:1175–1177CrossRefGoogle Scholar
  10. 10.
    Gould F, Clarke J, Heim C, Harvey PD, Majer M, Nemeroff CB (2012) The effects of child abuse and neglect on cognitive functioning in adulthood. J Psychiatr Res 46:500–506CrossRefGoogle Scholar
  11. 11.
    Nanni V, Uher R, Danese A (2012) Childhood maltreatment predicts unfavorable course of illness and treatment outcome in depression: a meta-analysis. Am J Psychiatry 169:141–151CrossRefGoogle Scholar
  12. 12.
    Hanson JL, Nacewicz BM, Sutterer MJ, Cayo AA, Schaefer SM, Rudolph KD, Shirtcliff EA, Pollak SD, Davidson RJ (2015) Behavioral problems after early life stress: contributions of the hippocampus and amygdala. Biol Psychiatry 77:314–323CrossRefGoogle Scholar
  13. 13.
    van Harmelen A-L, Hauber K, Gunther Moor B, Spinhoven P, Boon AE, Crone EA, Elzinga BM (2014) Childhood emotional maltreatment severity is associated with dorsal medial prefrontal cortex responsivity to social exclusion in young adults. PLoS One 9:e85107CrossRefGoogle Scholar
  14. 14.
    Mueller SC, Hardin MG, Korelitz K, Daniele T, Bemis J, Dozier M, Peloso E, Maheu FS, Pine DS, Ernst M (2012) Incentive effect on inhibitory control in adolescents with early-life stress: an antisaccade study. Child Abuse Negl 36:217–225CrossRefGoogle Scholar
  15. 15.
    Goff B, Gee DG, Telzer EH, Humphreys KL, Gabard-Durnam L, Flannery J, Tottenham N (2013) Reduced nucleus accumbens reactivity and adolescent depression following early-life stress. Neuroscience 249:129–138CrossRefGoogle Scholar
  16. 16.
    Mehta MA, Gore-Langton E, Golembo N, Colvert E, Williams SC, Sonuga-Barke E (2010) Hyporesponsive reward anticipation in the basal ganglia following severe institutional deprivation early in life. J Cogn Neurosci 22:2316–2325CrossRefGoogle Scholar
  17. 17.
    Gee DG, Gabard-Durnam LJ, Flannery J, Goff B, Humphreys KL, Telzer EH, Hare TA, Bookheimer SY, Tottenham N (2013) Early developmental emergence of human amygdala-prefrontal connectivity after maternal deprivation. Proc Natl Acad Sci U S A 110:15638–15643CrossRefGoogle Scholar
  18. 18.
    Sorrells SF, Caso JR, Munhoz CD, Sapolsky RM (2009) The stressed CNS: when glucocorticoids aggravate inflammation. Neuron 64:33–39CrossRefGoogle Scholar
  19. 19.
    de Pablos RM, Villaran RF, Arguelles S, Herrera AJ, Venero JL, Ayala A, Cano J, Machado A (2006) Stress increases vulnerability to inflammation in the rat prefrontal cortex. J Neurosci 26:5709–5719CrossRefGoogle Scholar
  20. 20.
    Gunn BG, Cunningham L, Cooper MA, Corteen NL, Seifi M, Swinny JD, Lambert JJ, Belelli D (2013) Dysfunctional astrocytic and synaptic regulation of hypothalamic glutamatergic transmission in a mouse model of early-life adversity: relevance to neurosteroids and programming of the stress response. J Neurosci 33:19534–19554CrossRefGoogle Scholar
  21. 21.
    Aisa B, Elizalde N, Tordera R, Lasheras B, Del Rio J, Ramirez MJ (2009) Effects of neonatal stress on markers of synaptic plasticity in the hippocampus: implications for spatial memory. Hippocampus 19:1222–1231CrossRefGoogle Scholar
  22. 22.
    Jutapakdeegul N, Afadlal S, Polaboon N, Phansuwan-Pujito P, Govitrapong P (2010) Repeated restraint stress and corticosterone injections during late pregnancy alter GAP-43 expression in the hippocampus and prefrontal cortex of rat pups. Int J Dev Neurosci 28:83–90CrossRefGoogle Scholar
  23. 23.
    McEwen BS, Gianaros PJ (2011) Stress- and allostasis-induced brain plasticity. Annu Rev Med 62:431–445CrossRefGoogle Scholar
  24. 24.
    Moghaddam B (1993) Stress preferentially increases extraneuronal levels of excitatory amino acids in the prefrontal cortex: comparison to hippocampus and basal ganglia. J Neurochem 60:1650–1657CrossRefGoogle Scholar
  25. 25.
    Munhoz CD, Sorrells SF, Caso JR, Scavone C, Sapolsky RM (2010) Glucocorticoids exacerbate lipopolysaccharide-induced signaling in the frontal cortex and hippocampus in a dose-dependent manner. J Neurosci 30:13690–13698CrossRefGoogle Scholar
  26. 26.
    Madrigal JL, Olivenza R, Moro MA, Lizasoain I, Lorenzo P, Rodrigo J, Leza JC (2001) Glutathione depletion, lipid peroxidation and mitochondrial dysfunction are induced by chronic stress in rat brain. Neuropsychopharmacology 24:420–429CrossRefGoogle Scholar
  27. 27.
    Manikandan S, Padma MK, Srikumar R, Jeya Parthasarathy N, Muthuvel A, Sheela Devi R (2006) Effects of chronic noise stress on spatial memory of rats in relation to neuronal dendritic alteration and free radical-imbalance in hippocampus and medial prefrontal cortex. Neurosci Lett 399:17–22CrossRefGoogle Scholar
  28. 28.
    Spiers JG, Chen HJ, Bradley AJ, Anderson ST, Sernia C, Lavidis NA (2013) Acute restraint stress induces rapid and prolonged changes in erythrocyte and hippocampal redox status. Psychoneuroendocrinology 38:2511–2519CrossRefGoogle Scholar
  29. 29.
    Nasselo AG, Machado C, Bastos JF, Felicio L (1998) Sudden darkness induces a high activity-lower anxiety state in male and female rats. Physiol Behav 63:451–454CrossRefGoogle Scholar
  30. 30.
    Uttara B, Singh AV, Zamboni P, Mahajan RT (2009) Oxidative stress and neurodegenerative diseases: a review of upstream and downstream antioxidant therapeutic options. Curr Neuropharmacol 7:65–74CrossRefGoogle Scholar
  31. 31.
    Yirmiya R, Goshen I (2011) Immune modulation of learning, memory, neural plasticity and neurogenesis. Brain Behav Immun 25:181–213CrossRefGoogle Scholar
  32. 32.
    Marik C, Felts PA, Bauer J, Lassmann H, Smith KJ (2007) Lesion genesis in a subset of patients with multiple sclerosis: a role for innate immunity? Brain 130:2800–2815CrossRefGoogle Scholar
  33. 33.
    Bilbo SD, Schwarz JM (2012) The immune system and developmental programming of brain and behavior. Front Neuroendocrinol 33:267–286CrossRefGoogle Scholar
  34. 34.
    Llorente-Berzal A, Assis MA, Rubino T, Zamberletti E, Marco EM, Parolaro D, Ambrosio E, Viveros MP (2013) Sex-dependent changes in brain CB1R expression and functionality and immune CB2R expression as a consequence of maternal deprivation and adolescent cocaine exposure. Pharmacol Res 74:23–33CrossRefGoogle Scholar
  35. 35.
    Pascual M, Blanco AM, Cauli O, Minarro J, Guerri C (2007) Intermittent ethanol exposure induces inflammatory brain damage and causes long-term behavioural alterations in adolescent rats. Eur J Neurosci 25:541–550CrossRefGoogle Scholar
  36. 36.
    Pascual M, Do Couto BR, Alfonso-Loeches S, Aguilar MA, Rodriguez-Arias M, Guerri C (2012) Changes in histone acetylation in the prefrontal cortex of ethanol-exposed adolescent rats are associated with ethanol-induced place conditioning. Neuropharmacology 62:2309–2319CrossRefGoogle Scholar
  37. 37.
    Rodriguez-Arias M, Maldonado C, Vidal-Infer A, Guerri C, Aguilar MA, Minarro J (2011) Intermittent ethanol exposure increases long-lasting behavioral and neurochemical effects of MDMA in adolescent mice. Psychopharmacology 218:429–442CrossRefGoogle Scholar
  38. 38.
    Khallouki F, Younos C, Soulimani R, Oster T, Charrouf Z, Spiegelhalder B, Bartsch H, Owen RW (2003) Consumption of argan oil (Morocco) with its unique profile of fatty acids, tocopherols, squalene, sterols and phenolic compounds should confer valuable cancer chemopreventive effects. Eur J Cancer Prev 12:67–75CrossRefGoogle Scholar
  39. 39.
    Meyer L, Caston J, Mensah-Nyagan AG (2006) Seasonal variation of the impact of a stressful procedure on open field behaviour and blood corticosterone in laboratory mice. Behav Brain Res 167:342–348CrossRefGoogle Scholar
  40. 40.
    Durand M, Berton O, Aguerre S, Edno L, Combourieu I, Mormede P, Chaouloff F (1999) Effects of repeated fluoxetine on anxiety-related behaviours, central serotonergic systems, and the corticotropic axis axis in SHR and WKY rats. Neuropharmacology 38:893–907CrossRefGoogle Scholar
  41. 41.
    Clénet F, Bouyon E, Hascoët M, Bourin M (2006) Light/dark cycle manipulation influences mice behaviour in the elevated plus maze. Behav Brain Res 166:140–149CrossRefGoogle Scholar
  42. 42.
    Porsolt RD, Anton G, Blavet N, Jalfre M (1978) Behavioural despair in rats: a new model sensitive to antidepressant treatments. Eur J Pharmacol 47:379–391CrossRefGoogle Scholar
  43. 43.
    Willner P (1997) Validity, reliability and utility of the chronic mild stress model of depression: a 10-year review and evaluation. Psychopharmacology 134:319–329CrossRefGoogle Scholar
  44. 44.
    Willner P, Muscat R, Papp M (1992) Chronic mild stress-induced anhedonia: a realistic animal model of depression. Neurosci Biobehav Rev 16:525–534CrossRefGoogle Scholar
  45. 45.
    Pothion S, Bizot JC, Trovero F, Belzung C (2004) Strain differences in sucrose preference and in the consequences of unpredictable chronic mild stress. Behav Brain Res 155:135–146CrossRefGoogle Scholar
  46. 46.
    Krishnan V, Han MH, Graham DL, Berton O, Renthal W, Russo SJ, Laplant Q, Graham A, Lutter M, Lagace DC, Ghose S, Reister R, Tannous P, Green TA, Neve RL, Chakravarty S, Kumar A, Eisch AJ, Self DW, Lee FS, Tamminga CA, Cooper DC, Gershenfeld HK, Nestler EJ (2007) Molecular adaptations underlying susceptibility and resistance to social defeat in brain reward regions. Cell 131:391–404CrossRefGoogle Scholar
  47. 47.
    Sierksma AS, van den Hove DL, Pfau F, Philippens M, Bruno O, Fedele E, Ricciarelli R, Steinbusch HW, Vanmierlo T, Prickaerts J (2014) Improvement of spatial memory function in APPswe/PS1dE9 mice after chronic inhibition of phosphodiesterase type 4D. Neuropharmacology 77:120–130CrossRefGoogle Scholar
  48. 48.
    Bevins RA, Besheer J (2006) Object recognition in rats and mice: a one-trial non-matching-to-sample learning task to study ‘recognition memory’. Nat Protoc 1:1306–1311CrossRefGoogle Scholar
  49. 49.
    Morris R (1984) Developments of a water-maze procedure for studying spatial learning in the rat. J Neurosci Methods 11:47–60CrossRefGoogle Scholar
  50. 50.
    Hodges H (1996) Maze procedures: the radial-arm and water maze compared. Brain Res Cogn Brain Res 3:167–181CrossRefGoogle Scholar
  51. 51.
    Owen AM, Evans AC, Petrides M (1996) Evidence for a two-stage model of spatial working memory processing within the lateral frontal cortex: a positron emission tomography study. Cereb Cortex 6:31–38CrossRefGoogle Scholar
  52. 52.
    Green LC, Tannenbaum SR, Goldman P (1981) Nitrate synthesis in the germfree and conventional rat. Science 212:56–58CrossRefGoogle Scholar
  53. 53.
    Esterbauer H (1993) Cytotoxicity and genotoxicity of lipid-oxidation products. Am J Clin Nutr 57:779S–785S; discussion 785S–786SCrossRefGoogle Scholar
  54. 54.
    Maehly AC, Chance B (1954) The assay of catalases and peroxidases. Methods Biochem Anal 1:357–424PubMedGoogle Scholar
  55. 55.
    Aebi H (1984) Catalase in vitro. Methods Enzymol 105:121–126CrossRefGoogle Scholar
  56. 56.
    Beauchamp C, Fridovich I (1971) Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal Biochem 44:276–287CrossRefGoogle Scholar
  57. 57.
    Howard C, Reed M (1998) Unbiased stereology. Three-dimensional measurement in microscopy. Oxford BIOS Scientific Publishers Limited, OxfordGoogle Scholar
  58. 58.
    Knudsen EI (2004) Sensitive periods in the development of the brain and behavior. J Cogn Neurosci 16:1412–1425CrossRefGoogle Scholar
  59. 59.
    Berrougui H, Alvarez de Sotomayor M, Perez-Guerrero C, Ettaib A, Hmamouchi M, Marhuenda E, Herrera MD (2004) Argan (Argania spinosa) oil lowers blood pressure and improves endothelial dysfunction in spontaneously hypertensive rats. Br J Nutr 92:921–929CrossRefGoogle Scholar
  60. 60.
    Mekhfi H, Belmekki F, Ziyyat A, Legssyer A, Bnouham M, Aziz M (2012) Antithrombotic activity of argan oil: an in vivo experimental study. Nutrition 28:937–941CrossRefGoogle Scholar
  61. 61.
    Bousalham R, Rhazali LJ, Harmouch A, Lotfi H, Benazzouz B, Hessni AE, Ouichou A, Akhouayri O, Mesfioui A (2015) Does argan oil supplementation affect metabolic parameters and behavior in wistar rats? Food Nutr Sci. Scholar
  62. 62.
    Weibel ER (1979) Stereological methods, Vol 1: Practical methods for biological morphometry. Zeitschrift für Allgemeine Mikrobiologie 21:630–630Google Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Hajar Benmhammed
    • 1
  • Samer El Hayek
    • 2
  • Inssaf Berkik
    • 1
  • Hicham Elmostafi
    • 1
  • Rim Bousalham
    • 1
  • Abdelhalem Mesfioui
    • 1
  • Ali Ouichou
    • 1
  • Aboubaker El Hessni
    • 1
  1. 1.Laboratory of Genetics, Neuroendocrinology, and Biotechnology, Department of Biology, Faculty of SciencesIbn Tofail UniversityKenitraMorocco
  2. 2.Department of Psychiatry, Faculty of MedicineAmerican University of BeirutBeirutLebanon

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