Advertisement

How Does Maternal Separation Affect the Cerebellum? Assessment of the Oxidative Metabolic Activity and Expression of the c-Fos Protein in Male and Female Rats

  • Alba Gutiérrez-Menéndez
  • María BanqueriEmail author
  • Marta Méndez
  • Jorge L. Arias
Original Paper

Abstract

Early life stress increases the risk of abnormal brain development, and it is associated with psychological disorders. Maternal separation is an established animal model of early life stress that produces changes in the development of the central nervous system. The objective of this study was to evaluate the effect of maternal separation on the rat cerebellum, both behaviourally and physiologically. We used 32 rats, males (n = 8) and females (n = 7), subjected to maternal separation for 21 days and a control group (9 males and 8 females). Spatial reference memory was assessed using the Morris water maze, and brain metabolic activity and the expression of an immediate early gene were determined, respectively, using the histochemical technique of cytochrome c oxidase and the immunocytochemistry of c-Fos. Results showed that both groups successfully performed the spatial memory task. Although there were no behavioural differences, the experimental group showed lower metabolic activity in the medial nucleus of the cerebellum, as well as fewer c-Fos-positive cells in the three deep nuclei of the cerebellum. These decreases could contribute to the emotional or behavioural impairments reported in maternal separation subjects.

Keywords

Cerebellum c-Fos Cytochrome c oxidase Early life stress Maternal separation 

Abbreviations

CF

control female group

CM

control male group

COX

cytochrome c oxidase

DN

dentate nucleus of the cerebellum

FN

fastigial nucleus of the cerebellum

INT

interposed nucleus of the cerebellum

LAT

lateral nucleus of the cerebellum

MED

medial nucleus of the cerebellum

MS

maternal separation

MS21M

maternal separation 21 male group

MS21F

maternal separation 21 female group

MWM

Morris water maze

PND

postnatal day

Notes

Funding Information

This research was supported by Projects Grants of the MINECO (Ministerio de Economía y Competitividad del Gobierno de España) PSI2017-90806-REDT, PSI2017-83893-R, PSI 2015-73111-EXP, AINDACE Foundation, and BES-2014- 070562 to M.B. The funding source had no role in the design of the study, data collection, analysis, interpretation, and writing of the present manuscript.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

Ethical Approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

References

  1. 1.
    Teicher MH, Andersen SL, Polcari A, Anderson CM, Navalta CP, Kim DM. The neurobiological consequences of early stress and childhood maltreatment. Neurosci Biobehav Rev. 2003;27:33–44.  https://doi.org/10.1016/S0149-7634(03)00007-1.CrossRefPubMedGoogle Scholar
  2. 2.
    Hedges DW, Woon FL. Early-life stress and cognitive outcome. Psychopharmacology. 2011;214:121–30.  https://doi.org/10.1007/s00213-010-2090-6.CrossRefPubMedGoogle Scholar
  3. 3.
    Anderson CM, Teicher MH, Polcari A, Renshaw PF. Abnormal T2 relaxation time in the cerebellar vermis of adults sexually abused in childhood: potential role of the vermis in stress-enhanced risk for drug abuse. Psychoneuroendocrinology. 2002;27:231–44.  https://doi.org/10.1016/S0306-4530(01)00047-6.CrossRefPubMedGoogle Scholar
  4. 4.
    O’Mahony SM, Marchesi JR, Scully P, Codling C, Ceolho AM, Quigley EMM, et al. Early life stress alters behavior, immunity, and microbiota in rats: implications for irritable bowel syndrome and psychiatric illnesses. Biol Psychiatry Soc Biol Psychiatry. 2009;65:263–7.  https://doi.org/10.1016/j.biopsych.2008.06.026.CrossRefGoogle Scholar
  5. 5.
    Oliveira SAA, Fontanelli BAFF, Stefanini MAA, Chuffa LGAGA, Teixeira GRR, Lizarte FSNSN, et al. Interaction of maternal separation on the UCh rat cerebellum. Microsc Res Tech. 2014;77:44–51.  https://doi.org/10.1002/jemt.22311.CrossRefPubMedGoogle Scholar
  6. 6.
    López-Gallardo M, Llorente R, Llorente-Berzal A, Marco EM, Prada C, Di Marzo V, et al. Neuronal and glial alterations in the cerebellar cortex of maternally deprived rats: gender differences and modulatory effects of two inhibitors of endocannabinoid inactivation. Dev Neurobiol. 2008;68:1429–40.  https://doi.org/10.1002/dneu.20672.CrossRefPubMedGoogle Scholar
  7. 7.
    Miki T, Yokoyama T, Kusaka T, Suzuki S, Ohta K, Warita K, et al. Early postnatal repeated maternal deprivation causes a transient increase in OMpg and BDNF in rat cerebellum suggesting precocious myelination. J Neurol Sci Elsevier BV. 2014;336:62–7.  https://doi.org/10.1016/j.jns.2013.10.007.CrossRefGoogle Scholar
  8. 8.
    Lalonde R, Botez MI. The cerebellum and learning processes in animals. Brain Res Rev. 1990;15:325–32.  https://doi.org/10.1016/j.brainres.2019.146358.CrossRefPubMedGoogle Scholar
  9. 9.
    Roque A, Lajud N, José Valdez J, Torner L. Early-life stress increases granule cell density in the cerebellum of male rats. Brain Res. 2019;1723:146358.  https://doi.org/10.1016/j.brainres.2019.146358.CrossRefPubMedGoogle Scholar
  10. 10.
    Amores-Villalba A, Mateos-Mateos R. Revisión de la neuropsicología del maltrato infantil: la neurobiología y el perfil neuropsicológico de las víctimas de abusos en la infancia. Psicol Educ Colegio Oficial de Psicólogos de Madrid. 2017;23:81–8.  https://doi.org/10.1016/j.pse.2017.05.006.CrossRefGoogle Scholar
  11. 11.
    R. L, C. S. The effects of cerebellar damage on maze learning in animals. Cerebellum. 2003;2:300–9.  https://doi.org/10.1080/14734220310017456.CrossRefGoogle Scholar
  12. 12.
    Fatemi SH, Reutiman TJ, Folsom TD, Sidwell RW. The role of cerebellar genes in pathology of autism and schizophrenia. Cerebellum. 2008;7:279–94.  https://doi.org/10.1007/s12311-008-0017-0.CrossRefPubMedGoogle Scholar
  13. 13.
    Bauer PM, Hanson JL, Pierson RK, Davidson RJ, Pollak SD. Cerebellar volume and cognitive functioning in children who experienced early deprivation. Biol Psychiatry. 2009;66:1100–6.  https://doi.org/10.1016/j.biopsych.2009.06.014.CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Leggio MG, Molinari M, Neri P, Graziano A, Mandolesi L, Petrosini L. Representation of actions in rats: the role of cerebellum in learning spatial performances by observation. Proc Natl Acad Sci. 2000;97:2320–5.  https://doi.org/10.1073/pnas.040554297.CrossRefPubMedGoogle Scholar
  15. 15.
    Hilber P. Influence of the cerebellum in anticipation and mental disorders. Anticip Med Cham: Springer International Publishing. 2017:137–46.  https://doi.org/10.1007/978-3-319-45142-8_8.Google Scholar
  16. 16.
    Banqueri M, Méndez M, Arias JL. Behavioral effects in adolescence and early adulthood in two length models of maternal separation in male rats. Behav Brain Res. 2017;324:77–86.  https://doi.org/10.1016/j.bbr.2017.02.006.CrossRefPubMedGoogle Scholar
  17. 17.
    Banqueri M, Méndez M, Arias JL. Spatial memory-related brain activity in normally reared and different maternal separation models in rats. Physiol Behav Elsevier. 2017;181:80–5.  https://doi.org/10.1016/j.physbeh.2017.09.007.CrossRefGoogle Scholar
  18. 18.
    Rubio S, Begega A, Méndez M, Méndez-López M, Arias JL. Similarities and differences between the brain networks underlying allocentric and egocentric spatial learning in rat revealed by cytochrome oxidase histochemistry. Neuroscience. 2012;223:174–82.  https://doi.org/10.1016/j.neuroscience.2012.07.066.CrossRefPubMedGoogle Scholar
  19. 19.
    Méndez M, Méndez-López M, López L, Aller MA, Arias J, Arias JL. Working memory impairment and reduced hippocampal and prefrontal cortex c-Fos expression in a rat model of cirrhosis. Physiol Behav. 2008;95:302–7.  https://doi.org/10.1016/j.physbeh.2008.06.013.CrossRefPubMedGoogle Scholar
  20. 20.
    Paxinos G, Watson C. The rat brain in stereotaxic coordinates. Elsevier Inc: Sixth Edition; 2007; 456.Google Scholar
  21. 21.
    Grace L, Hescham S, Kellaway LA, Bugarith K, Russell VA. Effect of exercise on learning and memory in a rat model of developmental stress. Metab Brain Dis. 2009;24:643–57.  https://doi.org/10.1007/s11011-009-9162-5.CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Moreno-Rius J. The cerebellum under stress. Front Neuroendocrinol Elsevier. 2019;54:100774.  https://doi.org/10.1016/j.yfrne.2019.100774.CrossRefGoogle Scholar
  23. 23.
    Wang Q, Shao F, Wang W. Maternal separation produces alterations of forebrain brain-derived neurotrophic factor expression in differently aged rats. Front Mol Neurosci. 2015;8:1–8.  https://doi.org/10.3389/fnmol.2015.00049.CrossRefGoogle Scholar
  24. 24.
    Wilber AA, Wellman CL. Neonatal maternal separation-induced changes in glucocorticoid receptor expression in posterior interpositus interneurons but not projection neurons predict deficits in adult eyeblink conditioning. Neurosci Lett. 2009;460:214–8.  https://doi.org/10.1016/j.ijdevneu.2009.08.001.CrossRefPubMedGoogle Scholar
  25. 25.
    Wang Q, Li M, Du W, Shao F, Wang W. The different effects of maternal separation on spatial learning and reversal learning in rats. Behav Brain Res Elsevier BV. 2015;280:16–23.  https://doi.org/10.1016/j.bbr.2014.11.040.CrossRefGoogle Scholar
  26. 26.
    Aisa B, Tordera R, Lasheras B, Del Río J, Ramírez MJ. Cognitive impairment associated to HPA axis hyperactivity after maternal separation in rats. Psychoneuroendocrinology. 2007;32:256–66.  https://doi.org/10.1016/j.psyneuen.2006.12.013.CrossRefPubMedGoogle Scholar
  27. 27.
    Cao X, Huang S, Cao J, Chen T, Zhu P, Zhu R, et al. The timing of maternal separation affects Morris water maze performance and long-term potentiation in male rats. Dev Psychobiol. 2014;56:1102–9.  https://doi.org/10.1002/dev.21130.CrossRefPubMedGoogle Scholar
  28. 28.
    Wilber AA, Wellman CL. Neonatal maternal separation alters the development of glucocorticoid receptor expression in the interpositus nucleus of the cerebellum. Int J Dev Neurosci. 2009;27:649–54.  https://doi.org/10.1016/j.neulet.2009.05.076.CrossRefPubMedGoogle Scholar
  29. 29.
    Foti F, Mandolesi L, Cutuli D, Laricchiuta D, De Bartolo P, Gelfo F, et al. Cerebellar damage loosens the strategic use of the spatial structure of the search space. Cerebellum. 2010;9:29–41.  https://doi.org/10.1007/s12311-009-0134-4.CrossRefPubMedGoogle Scholar
  30. 30.
    Molinari M, Leggio MG. Cerebellar information processing and visuospatial functions. Cerebellum. 2007;6:214–20.  https://doi.org/10.1080/14734220701230870.CrossRefPubMedGoogle Scholar
  31. 31.
    Zhu X, Li T, Peng S, Ma X, Chen X, Zhang X. Maternal deprivation-caused behavioral abnormalities in adult rats relate to a non-methylation-regulated D2 receptor levels in the nucleus accumbens. Behav Brain Res. 2010;209:281–8.  https://doi.org/10.1016/j.bbr.2010.02.005.CrossRefPubMedGoogle Scholar
  32. 32.
    Jie HJ, Jun ZZ, Shan LS, Jun XG, Rong ZX, Teng GJ, et al. Hippocampal neurochemistry is involved in the behavioural effects of neonatal maternal separation and their reversal by post-weaning environmental enrichment: a magnetic resonance study. Behav Brain Res. 2011;217:122–7.  https://doi.org/10.1016/j.bbr.2010.10.014.CrossRefGoogle Scholar
  33. 33.
    Spivey JM, Padilla E, Shumake JD, Gonzalez-Lima F. Effects of maternal separation, early handling, and gonadal sex on regional metabolic capacity of the preweanling rat brain. Brain Res Elsevier BV. 2011;1367:198–206.  https://doi.org/10.1016/j.brainres.2010.10.038.CrossRefGoogle Scholar
  34. 34.
    Zhang X-Y, Wang J-J, Zhu J-N. Cerebellar fastigial nucleus: from anatomic construction to physiological functions. Cerebellum Ataxias. 2016;3:9.  https://doi.org/10.1186/s40673-016-0047-1.CrossRefPubMedPubMedCentralGoogle Scholar
  35. 35.
    Golanov E, Regnier-Golanov A, Britz G. Integrity of cerebellar fastigial nucleus intrinsic neurons is critical for the global ischemic preconditioning. Brain Sci. 2017;7:121.  https://doi.org/10.3390/brainsci7100121.CrossRefPubMedCentralGoogle Scholar
  36. 36.
    Zhang L, Zhao M, Sui R-B. Cerebellar fastigial nucleus electrical stimulation alleviates depressive-like behaviors in post-stroke depression rat model and potential mechanisms. Cell Physiol Biochem. 2017;41:1403–12.  https://doi.org/10.1186/s40673-016-0047-1.CrossRefPubMedGoogle Scholar
  37. 37.
    Huguet G, Kadar E, Temel Y, Lim LW. Electrical stimulation normalizes c-Fos expression in the deep cerebellar nuclei of depressive-like rats: implication of antidepressant activity. Cerebellum. 2017;16:398–410.  https://doi.org/10.1007/s12311-016-0812-y.CrossRefPubMedGoogle Scholar
  38. 38.
    Coffman KA, Dum RP, Strick PL. Cerebellar vermis is a target of projections from the motor areas in the cerebral cortex. Proc Natl Acad Sci. 2011;108:16068–73.  https://doi.org/10.1073/pnas.1107904108.CrossRefPubMedGoogle Scholar
  39. 39.
    Ketter TA, Kimbrell TA, George MS, Dunn RT, Speer AM, Benson BE, et al. Effects of mood and subtype on cerebral glucose metabolism in treatment-resistant bipolar disorder. Biol Psychiatry. 2001;49:97–109.  https://doi.org/10.1016/S0006-3223(00)00975-6.CrossRefPubMedGoogle Scholar
  40. 40.
    Heim C, Binder EB. Current research trends in early life stress and depression: review of human studies on sensitive periods, gene–environment interactions, and epigenetics. Exp Neurol Elsevier Inc. 2012;233:102–11.  https://doi.org/10.1016/j.expneurol.2011.10.032.CrossRefGoogle Scholar
  41. 41.
    Peng Y-P, Qiu Y-H, Qiu J, Wang J-J. Cerebellar interposed nucleus lesions suppress lymphocyte function in rats. Brain Res Bull. 2006;71:10–7.  https://doi.org/10.1016/j.brainresbull.2006.07.017.CrossRefPubMedGoogle Scholar
  42. 42.
    Kovács KJ. Invited review c-Fos as a transcription factor: a stressful (re)view from a functional map. Neurochem Int. 1998;33:287–97.  https://doi.org/10.1016/S0197-0186(98)00023-0.CrossRefPubMedGoogle Scholar

Copyright information

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

Authors and Affiliations

  1. 1.Department of Psychology, Laboratory of NeuroscienceUniversity of OviedoOviedoSpain
  2. 2.Instituto de Neurociencias del Principado de Asturias (INEUROPA)OviedoSpain

Personalised recommendations