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Epigenetic programming by maternal behavior


Here we report that increased pup licking and grooming (LG) and arched-back nursing (ABN) by rat mothers altered the offspring epigenome at a glucocorticoid receptor (GR) gene promoter in the hippocampus. Offspring of mothers that showed high levels of LG and ABN were found to have differences in DNA methylation, as compared to offspring of 'low-LG-ABN' mothers. These differences emerged over the first week of life, were reversed with cross-fostering, persisted into adulthood and were associated with altered histone acetylation and transcription factor (NGFI-A) binding to the GR promoter. Central infusion of a histone deacetylase inhibitor removed the group differences in histone acetylation, DNA methylation, NGFI-A binding, GR expression and hypothalamic-pituitary-adrenal (HPA) responses to stress, suggesting a causal relation among epigenomic state, GR expression and the maternal effect on stress responses in the offspring. Thus we show that an epigenomic state of a gene can be established through behavioral programming, and it is potentially reversible.

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Change history

  • 27 July 2004

    added footnote and footnote references to figures within text; updated Figure 1; corrected online date made to issue version of PDF


  1. 1.

    *Note: In the version of this article originally published online, in Figure 1, the label of the y axis was omitted: it should read "C-methylation (%)". In Figure 2, the panels were misidentified: the top two Southern blot panels in the immunoprecipitation analysis data should be identified as "a", the bottom two Southern blot panels as "b" and the graph as "c". In Figure 3, the legend text referred incorrectly to the panels: the legend should refer to the blots (beginning in the second sentence) as "a" and the graph (last sentence) as "b". In Figure 5, the panels were misidentified: the western blots and the graph immediately below them should be identified as "a" and the graph of corticosterone response as "b". In addition, in panel b, the units were listed incorrectly on the y axis: the correct units should be "μg/dl". In the Methods section, in the description of "Sodium bisufite mapping", the description of the thermocycler protocol listed the annealing temperature incorrectly: the annealing temperature should be 56 °C.


  1. 1

    Agrawal, A.A. Phenotypic plasticity in the interactions and evolution of species. Science 294, 321–326 (2001).

  2. 2

    Rossiter, M.C. Maternal Effects as Adaptations (eds. Fox, T.A. & Mousseau, C.W.) (Oxford University Press, London, 1999).

  3. 3

    Levine, S. The ontogeny of the hypothalamic-pituitary-adrenal axis. The influence of maternal factors. Ann. NY Acad. Sci. 746, 275–293 (1994).

  4. 4

    Fleming, A.S., O'Day, D.H. & Kraemer, G.W. Neurobiology of mother-infant interactions: experience and central nervous system plasticity across development and generations. Neurosci. Biobehav. Rev. 23, 673–685 (1999).

  5. 5

    Meaney, M.J. Maternal care, gene expression, and the transmission of individual differences in stress reactivity across generations. Annu. Rev. Neurosci. 24, 1161–1192 (2001).

  6. 6

    Stern, J.M. Offspring-induced nurturance: animal-human parallels. Dev. Psychobiol. 31, 19–37 (1997).

  7. 7

    Liu, D. et al. Maternal care, hippocampal glucocorticoid receptors, and hypothalamic-pituitary-adrenal responses to stress. Science 277, 1659–1662 (1997).

  8. 8

    Caldji, C. et al. Maternal care during infancy regulates the development of neural systems mediating the expression of fearfulness in the rat. Proc. Natl. Acad. Sci. USA 95, 5335–5340 (1998).

  9. 9

    Francis, D., Diorio, J., Liu, D. & Meaney, M.J. Nongenomic transmission across generations of maternal behavior and stress responses in the rat. Science 286, 1155–1158 (1999).

  10. 10

    Myers, M.M., Brunelli, S.A., Shair, H.N., Squire, J.M. & Hofer, M.A. Relationships between maternal behavior of SHR and WKY dams and adult blood pressures of cross-fostered F1 pups. Dev. Psychobiol. 22, 55–67 (1989).

  11. 11

    De Kloet, E.R., Vreugdenhil, E., Oitzl, M.S. & Joels, M. Brain corticosteroid receptor balance in health and disease. Endocr. Rev. 19, 269–301 (1998).

  12. 12

    Meaney, M.J., Aitken, D.H., Viau, V., Sharma, S. & Sarrieau, A. Neonatal handling alters adrenocortical negative feedback sensitivity and hippocampal type II glucocorticoid receptor binding in the rat. Neuroendocrinology 50, 597–604 (1989).

  13. 13

    Weaver, I.C. et al. Early environmental regulation of hippocampal glucocorticoid receptor gene expression: characterization of intracellular mediators and potential genomic target sites. Mol. Cell. Endocrinol. 185, 205–218 (2001).

  14. 14

    Meaney, M.J. et al. Postnatal handling increases the expression of cAMP-inducible transcription factors in the rat hippocampus: the effects of thyroid hormones and serotonin. J. Neurosci. 20, 3926–3935 (2000).

  15. 15

    Laplante, P., Diorio, J. & Meaney, M.J. Serotonin regulates hippocampal glucocorticoid receptor expression via a 5-HT7 receptor. Brain Res. Dev. Brain Res. 139, 199–203 (2002).

  16. 16

    McCormick, J.A. et al. 5′-heterogeneity of glucocorticoid receptor messenger RNA is tissue specific: differential regulation of variant transcripts by early-life events. Mol. Endocrinol. 14, 506–517 (2000).

  17. 17

    Kadonaga, J.T. Eukaryotic transcription: an interlaced network of transcription factors and chromatin-modifying machines. Cell 92, 307–313 (1998).

  18. 18

    Razin, A. CpG methylation, chromatin structure and gene silencing- a three-way connection. Embo J. 17, 4905–4908 (1998).

  19. 19

    Keshet, I., Yisraeli, J. & Cedar, H. Effect of regional DNA methylation on gene expression. Proc. Natl. Acad. Sci. USA 82, 2560–2564 (1985).

  20. 20

    Razin, A. & Cedar, H. Distribution of 5-methylcytosine in chromatin. Proc. Natl. Acad. Sci. USA 74, 2725–2728 (1977).

  21. 21

    Clark, S.J., Harrison, J., Paul, C.L. & Frommer, M. High sensitivity mapping of methylated cytosines. Nucleic Acids Res. 22, 2990–2997 (1994).

  22. 22

    Frommer, M. et al. A genomic sequencing protocol that yields a positive display of 5-methylcytosine residues in individual DNA strands. Proc. Natl. Acad. Sci. USA 89, 1827–1831 (1992).

  23. 23

    Cintra, A. et al. Mapping and computer assisted morphometry and microdensitometry of glucocorticoid receptor immunoreactive neurons and glial cells in the rat central nervous system. Neuroscience 62, 843–897 (1994).

  24. 24

    Cintra, A. et al. Glial and neuronal glucocorticoid receptor immunoreactive cell populations in developing, adult, and aging brain. Ann. NY Acad. Sci. 746, 42–63 (1994).

  25. 25

    Brinton, R.D., Yamazaki, R., Gonzalez, C.M., O'Neill, K. & Schreiber, S.S. Vasopressin-induction of the immediate early gene, NGFI-A, in cultured hippocampal glial cells. Brain Res. Mol. Brain Res. 57, 73–85 (1998).

  26. 26

    Sapienza, C. Parental imprinting of genes. Sci. Am. 263, 52–60 (1990).

  27. 27

    Hershko, A.Y., Kafri, T., Fainsod, A. & Razin, A. Methylation of HoxA5 and HoxB5 and its relevance to expression during mouse development. Gene 302, 65–72 (2003).

  28. 28

    Meaney, M.J. et al. Early environmental regulation of forebrain glucocorticoid receptor gene expression: implications for adrenocortical responses to stress. Dev. Neurosci. 18, 49–72 (1996).

  29. 29

    Roth, S.Y., Denu, J.M. & Allis, C.D. Histone acetyltransferases. Annu. Rev. Biochem. 70, 81–120 (2001).

  30. 30

    Cervoni, N. & Szyf, M. Demethylase activity is directed by histone acetylation. J. Biol. Chem. 276, 40778–40787 (2001).

  31. 31

    Milbrandt, J. A nerve growth factor-induced gene encodes a possible transcriptional regulatory factor. Science 238, 797–799 (1987).

  32. 32

    Champagne, F.A., Francis, D.D., Mar, A. & Meaney, M.J. Variations in maternal care in the rat as a mediating influence for the effects of environment on development. Physiol. Behav. 79, 359–371 (2003).

  33. 33

    Crane-Robinson, C., Myers, F.A., Hebbes, T.R., Clayton, A.L. & Thorne, A.W. Chromatin immunoprecipitation assays in acetylation mapping of higher eukaryotes. Methods Enzymol. 304, 533–547 (1999).

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These studies were supported by a grant from the Canadian Institutes for Health Research (CIHR) to M.J.M. and M.S. and from the National Cancer Institute of Canada to M.S. M.J.M. is supported by a CIHR Senior Scientist award and the project was supported by a Distinguished Investigator Award (M.J.M.) from the National Alliance for Research on Schizophrenia and Affective Disorders (NARSAD).

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The authors declare no competing financial interests.

Correspondence to Moshe Szyf or Michael J Meaney.

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Figure 1: Maternal care alters cytosine methylation of GR promoter.
Figure 2: Chromatin immunoprecipitation analysis of the association between histone H3-K9 acetylation and NGFI-A binding to the exon 17 GR sequence in hippocampal tissue from adult offspring of high- and low-LG-ABN mothers (n = 4 animals/group).
Figure 3: HDAC inhibitior (TSA) eliminates maternal effect on histone acetylation and NGFI-A binding.
Figure 4: TSA effects on cytosine methylation.
Figure 5: TSA eliminates the maternal effect on hippocampal GR expression and HPA responses to stress.