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Molecular Neurobiology

, Volume 55, Issue 1, pp 567–582 | Cite as

Voluntary Physical Exercise Induces Expression and Epigenetic Remodeling of VegfA in the Rat Hippocampus

  • Christina A. E. Sølvsten
  • Frank de Paoli
  • Jane H. Christensen
  • Anders L. NielsenEmail author
Article

Abstract

A healthy lifestyle, including regular physical exercise, is generally believed to improve cognitive function and enhance neurogenesis. Such physical exercise-induced effects are associated with increased brain expression of neurotrophic and growth factors. In the present study, we investigated Bdnf, Igf-1, Fgf-2, Egf, and VegfA messenger RNA (mRNA) expression levels in the male rat hippocampus and frontal cortex after 2 weeks of voluntary physical exercise. Whereas the expression of Fgf-2 was upregulated in the hippocampus and prefrontal cortex by physical exercise, the expression levels of Bdnf transcript 1, Bdnf transcript 4, Igf-1, and VegfA were upregulated only in the hippocampus. We focused our subsequent analyses on the VegfA gene, which encodes vascular endothelial growth factor, a signaling molecule important for angiogenesis, vasculogenesis, and neurogenesis. To study the epigenetic mechanisms involved in the physical exercise-mediated induction of VegfA expression, we used oxidative and non-oxidative bisulfite pyrosequencing to analyze VegfA promoter DNA methylation and DNA hydroxymethylation. We observed discrete DNA hypomethylation at specific CpG sites in rats that engaged in physical exercise relative to sedentary rats. This is exemplified by a CpG site located within a VegfA promoter Sp1/Sp3 transcription factor recognition element. DNA hydroxymethylation was present at the VegfA promoter, but no differences in DNA hydroxymethylation were observed in rats that engaged in physical exercise relative to sedentary rats. Moreover, we observed increased Tet1 and decreased Dnmt3b mRNA expression in the hippocampi of rats that engaged in physical exercise. The presented results substantiate the involvement of epigenetics as a mediator of the beneficial effects of physical exercise and point to the importance of analyzing factors beyond Bdnf to delineate the mechanisms behind the functional impacts of physical exercise in mediating benefits to the brain.

Keywords

Physical exercise Neurotrophic factors Epigenetics Bdnf Gene regulation 

Abbreviations

Bdnf

Brain-derived neurotrophic factor

ChIP

Chromatin immunoprecipitation

CpG-I

CpG island

Dnmts

DNA methyltransferases

Ngf

Nerve growth factor

Nt-3

Neurotrophin-3

Nt-4

Neurotrophin-4

Hif-1

Hypoxia-inducible factor 1

Igf-1

Insulin-like growth factor-1

Fgf-2

Fibroblast growth factor-2

VegfA

Vascular endothelial growth factor A

SEM

Standard of mean

Notes

Acknowledgements

This work was supported by the Toyota Foundation and the Lundbeck Foundation (R100-A9606). We thank Tina Fuglsang Daugaard for excellent technical assistance.

Author Contributions

CS, FDP, JHC, and ALN conceived and designed the study. CS performed the biological experiments. All authors contributed to the writing of the manuscript and approved the final version of the manuscript.

Compliance with Ethical Standards

Conflict of Interest

The authors declare no conflicts of interest.

Supplementary material

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Table S1 (DOCX 17 kb)
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Table S2 (DOCX 16 kb)
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Table S3 (DOCX 17 kb)
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Table S4 (DOCX 15 kb)
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Table S5 (DOCX 19 kb)
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Fig S1

Schematic illustration of the structure of the rat Bdnf gene. Nine Bdnf promoters are identified with promoters 1 to 8 driving the transcription of Bdnf mRNA containing one of the eight alternative non-coding 5′ exons (exons 1–8) all spliced to the common protein coding 3′ exon 9. In addition, transcription can be initiated upstream of exon 9 with a resulting exon 9a which includes a 5′ sequence extension of the exon 9 sequence. (GIF 4 kb)

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Fig S2

Grouping of exercised animals according to cumulative running distance. The rats from the exercised group were subdivided into three groups with four rats in each group. Rats in group 1 (G1) had run between 7.25 and 9.16 km in total, rats in group 2 (G2) had run between 14.44 and 26.84 km, and rats in group 3 (G3) had run between 46.31 and 55.92 km. To the right are illustrated G1, G2, and G3 classification according to the cumulative running distance. (GIF 9 kb)

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High resolution image (EPS 639 kb)
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Fig S3

Neurotrophic factor mRNA expression analysis in hippocampus of physically exercised animals subgrouped accordingly to cumulative running distance. The mean mRNA expression level in the sedentary group was tested against the mean expression level in each of the three exercise subgroups G1, G2, and G3. Expression data were normalized to the expression of the reference genes Ppia and Rpl13A, and the expression level in sedentary rats normalized to value one. Values represent the mean + SEM. Expression differences between the exercised subgroups of rats and the sedentary rats were analyzed by one-way ANOVA followed by Bonferroni’s multiple comparison test with *P ≤ 0.05 and **P ≤ 0.01. (GIF 36 kb)

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Fig S4

Neurotrophic factor mRNA expression analysis in frontal cortex of exercised rats grouped accordingly to cumulative running distance. See legend to Fig. S3 for details. (GIF 34 kb)

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Fig S5

Expression analysis of growth factors Igf-1, VegfA, Fgf-2, and Egf mRNA in hippocampus of exercised rats grouped accordingly to cumulative running distance. See legend to Fig. S3 for details. (GIF 18 kb)

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Fig S6

Expression analysis of growth factors Igf-1, VegfA, Fgf-2, and Egf mRNA in frontal cortex of exercised rats grouped accordingly to cumulative running distance. See legend to Fig. S3 for details. (GIF 18 kb)

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Fig S7

Expression analysis of Dnmt3a, Dnmt3b, Dnmt1, and Tet1 mRNA in hippocampus of exercised rats grouped accordingly to cumulative running distance. See legend to Fig. S3 for details. (GIF 18 kb)

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Fig S8

Immunohistochemical staining with hematoxylin (blue) and VegfA-antibody (brown) of the dentate gyrus of physically exercised rat number 10 (19.6 km in total) (a, b) and a control sedentary rat (c, d). Immunohistochemistry was performed on hippocampus cryosections. Five-micrometer-thick sections were mounted on coated glass slides. The tissue sections were fixed in Lillies fixative. The slides were treated with peroxidase block (DAKO) for 5 min and blocked with bovine serum albumin for 30 min. Immunohistochemical detection of VegfA was performed using the EnVision+ System-HRP. To detect VegfA protein, rabbit anti-rat VegfA antibody (ab46157) was used in a dilution of 1:600 and incubated ON at 4 °C. The sections were counterstained with Mayer’s hematoxylin solution. The slides were finally cover slipped with mounting medium (DAKO) and analyzed by a Leica DM 2500 microscope using Leica IM50 4.0 software. Magnification is ×10 (a, c) and ×40 (b, d) with the latter representing the red squared boxes (a, c) (GIF 145 kb)

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References

  1. 1.
    Gomez-Pinilla F, Vaynman S, Ying Z (2008) Brain-derived neurotrophic factor functions as a metabotrophin to mediate the effects of exercise on cognition. Eur J Neurosci 28:2278–2287PubMedPubMedCentralCrossRefGoogle Scholar
  2. 2.
    Pang TYC, Hannan AJ (2013) Enhancement of cognitive function in models of brain disease through environmental enrichment and physical activity. Neuropharmacology 64:515–528PubMedCrossRefGoogle Scholar
  3. 3.
    Thomas AG, Dennis A, Bandettini PA, Johansen-Berg H (2012) The effects of aerobic activity on brain structure. Front Psychol 3:86PubMedPubMedCentralCrossRefGoogle Scholar
  4. 4.
    van Praag H, Kempermann G, Gage FH (1999b) Running increases cell proliferation and neurogenesis in the adult mouse dentate gyrus. Nat Neurosci 2:266–270PubMedCrossRefGoogle Scholar
  5. 5.
    Vaynman S, Gomez-Pinilla F (2005) License to run: exercise impacts functional plasticity in the intact and injured central nervous system by using neurotrophins. Neurorehabil. Neurol Rep 19:283–295Google Scholar
  6. 6.
    Farmer J, Zhao X, van Praag H, Wodtke K, Gage FH, Christie BR (2004) Effects of voluntary exercise on synaptic plasticity and gene expression in the dentate gyrus of adult male Sprague-Dawley rats in vivo. Neuroscience 124:71–79PubMedCrossRefGoogle Scholar
  7. 7.
    Molteni R, Ying Z, Gómez-Pinilla F (2002) Differential effects of acute and chronic exercise on plasticity-related genes in the rat hippocampus revealed by microarray. Eur J Neurosci 16:1107–1116PubMedCrossRefGoogle Scholar
  8. 8.
    Vaynman SS, Ying Z, Yin D, Gomez-Pinilla F (2006) Exercise differentially regulates synaptic proteins associated to the function of BDNF. Brain Res 1070:124–130PubMedCrossRefGoogle Scholar
  9. 9.
    Abel JL, Rissman EF (2013) Running-induced epigenetic and gene expression changes in the adolescent brain. Int J Dev Neurosci 31:382–390PubMedCrossRefGoogle Scholar
  10. 10.
    Adlard PA, Perreau VM, Cotman CW (2005) The exercise-induced expression of BDNF within the hippocampus varies across life-span. Neurobiol Aging 26:511–520PubMedCrossRefGoogle Scholar
  11. 11.
    Ding Q, Vaynman S, Akhavan M, Ying Z, Gomez-Pinilla F (2006) Insulin-like growth factor I interfaces with brain-derived neurotrophic factor-mediated synaptic plasticity to modulate aspects of exercise-induced cognitive function. Neuroscience 140:823–833PubMedCrossRefGoogle Scholar
  12. 12.
    Fang ZH, Lee CH, Seo MK, Cho H, Lee JG, Lee BJ, Park SW, Kim YH (2013) Effect of treadmill exercise on the BDNF-mediated pathway in the hippocampus of stressed rats. Neurosci Res 76:187–194PubMedCrossRefGoogle Scholar
  13. 13.
    Gomes da Silva S, Unsain N, Mascó DH, Toscano-Silva M, de Amorim HA, Silva Araújo BH, Simões PSR, Naffah-Mazzacoratti Da MG et al (2012) Early exercise promotes positive hippocampal plasticity and improves spatial memory in the adult life of rats. Hippocampus 22:347–358PubMedCrossRefGoogle Scholar
  14. 14.
    Vaynman S, Ying Z, Gomez-Pinilla F (2004) Hippocampal BDNF mediates the efficacy of exercise on synaptic plasticity and cognition. Eur J Neurosci 20:2580–2590PubMedCrossRefGoogle Scholar
  15. 15.
    Reichardt LF (2006) Neurotrophin-regulated signalling pathways. Philos Trans R Soc Lond Ser B Biol Sci 361:1545–1564CrossRefGoogle Scholar
  16. 16.
    Huang EJ, Reichardt LF (2001) Neurotrophins: roles in neuronal development and function. Annu Rev Neurosci 24:677–736PubMedPubMedCentralCrossRefGoogle Scholar
  17. 17.
    Lewin GR, Barde Y-A (1996) Physiology of the neurotrophins. Annu Rev Neurosci 19:289–317PubMedCrossRefGoogle Scholar
  18. 18.
    Lu B, Pang PT, Woo NH (2005) The yin and yang of neurotrophin action. Nat Rev Neurosci 6:603–614PubMedCrossRefGoogle Scholar
  19. 19.
    Aid T, Kazantseva A, Piirsoo M, Palm K (2007) Mouse and rat BDNF Gene structure and expression revisited. J Neurosci Res 535:525–535CrossRefGoogle Scholar
  20. 20.
    Chen WG, Chang Q, Lin Y, Meissner A, West AE, Griffith EC, Jaenisch R, Greenberg ME (2003) Derepression of BDNF transcription involves calcium-dependent phosphorylation of MeCP2. Science 302:885–889PubMedCrossRefGoogle Scholar
  21. 21.
    Gomez-Pinilla F, Zhuang Y, Feng J, Ying Z, Fan G (2011) Exercise impacts brain-derived neurotrophic factor plasticity by engaging mechanisms of epigenetic regulation. Eur J Neurosci 33:383–390PubMedCrossRefGoogle Scholar
  22. 22.
    Lu B (2003) BDNF and activity-dependent synaptic modulation. Learn Mem 10:86–98PubMedPubMedCentralCrossRefGoogle Scholar
  23. 23.
    Gómez-Pinilla F, Dao L, So V (1997) Physical exercise induces FGF-2 and its mRNA in the hippocampus. Brain Res 764:1–8PubMedCrossRefGoogle Scholar
  24. 24.
    Lee E, Son H (2009) Adult hippocampal neurogenesis and related neurotrophic factors. BMB Rep 42:239–244PubMedCrossRefGoogle Scholar
  25. 25.
    Masi G, Brovedani P (2011) The hippocampus, neurotrophic factors and depression: possible implications for the pharmacotherapy of depression. CNS Drugs 25:913–931PubMedCrossRefGoogle Scholar
  26. 26.
    Schmidt HD, Duman RS (2007) The role of neurotrophic factors in adult hippocampal neurogenesis, antidepressant treatments and animal models of depressive-like behavior. Behav Pharmacol 18:391–418PubMedCrossRefGoogle Scholar
  27. 27.
    Millauer B, Wizigmann-Voos S, Schnürch H, Martinez R, Møller NP, Risau W, Ullrich A (1993) High affinity VEGF binding and developmental expression suggest Flk-1 as a major regulator of vasculogenesis and angiogenesis. Cell 72:835–846PubMedCrossRefGoogle Scholar
  28. 28.
    Jin K, Zhu Y, Sun Y, Mao XO, Xie L, Greenberg DA (2002) Vascular endothelial growth factor (VEGF) stimulates neurogenesis in vitro and in vivo. Proc Natl Acad Sci U S A 99:11946–11950PubMedPubMedCentralCrossRefGoogle Scholar
  29. 29.
    Jin KL, Mao XO, Greenberg DA (2000) Vascular endothelial growth factor: direct neuroprotective effect in in vitro ischemia. Proc Natl Acad Sci U S A 97:10242–10247PubMedPubMedCentralCrossRefGoogle Scholar
  30. 30.
    Matsuzaki H, Tamatani M, Yamaguchi A, Namikawa K, Kiyama H, Vitek MP, Mitsuda N, Tohyama M (2001) Vascular endothelial growth factor rescues hippocampal neurons from glutamate-induced toxicity: signal transduction cascades. FASEB J 15:1218–1220PubMedCrossRefGoogle Scholar
  31. 31.
    Ogunshola OO, Antic A, Donoghue MJ, Fan S-Y, Kim H, Stewart WB, Madri JA, Ment LR (2002) Paracrine and autocrine functions of neuronal vascular endothelial growth factor (VEGF) in the central nervous system. J Biol Chem 277:11410–11415PubMedCrossRefGoogle Scholar
  32. 32.
    Fabel K, Fabel K, Tam B, Kaufer D, Baiker A, Simmons N, Kuo CJ, Palmer TD (2003) VEGF is necessary for exercise-induced adult hippocampal neurogenesis. Eur J Neurosci 18:2803–2812PubMedCrossRefGoogle Scholar
  33. 33.
    Kiuchi T, Lee H, Mikami T (2012) Regular exercise cures depression-like behavior via VEGF-Flk-1 signaling in chronically stressed mice. Neuroscience 207:208–217PubMedCrossRefGoogle Scholar
  34. 34.
    Lambrechts D, Carmeliet P (2006) VEGF at the neurovascular interface: therapeutic implications for motor neuron disease. Biochim Biophys Acta 1762:1109–1121PubMedCrossRefGoogle Scholar
  35. 35.
    Storkebaum E, Carmeliet P (2004) VEGF: a critical player in neurodegeneration. J Clin Invest 113:14–18PubMedPubMedCentralCrossRefGoogle Scholar
  36. 36.
    Zacchigna S, Lambrechts D, Carmeliet P (2008) Neurovascular signalling defects in neurodegeneration. Nat Rev Neurosci 9:169–181PubMedCrossRefGoogle Scholar
  37. 37.
    Forsythe JA, Jiang BH, Iyer NV, Agani F, Leung SW, Koos RD, Semenza GL (1996) Activation of vascular endothelial growth factor gene transcription by hypoxia-inducible factor 1. Mol Cell Biol 16:4604–4613PubMedPubMedCentralCrossRefGoogle Scholar
  38. 38.
    Pagès G, Pouysségur J (2005) Transcriptional regulation of the vascular endothelial growth factor gene—a concert of activating factors. Cardiovasc Res 65:564–573PubMedCrossRefGoogle Scholar
  39. 39.
    Arendt T (2009) Synaptic degeneration in Alzheimer’s disease. Acta Neuropathol 118:167–179PubMedCrossRefGoogle Scholar
  40. 40.
    Tsankova N, Renthal W, Kumar A, Nestler EJ (2007) Epigenetic regulation in psychiatric disorders. Nat Rev Neurosci 8:355–367PubMedCrossRefGoogle Scholar
  41. 41.
    Bird A (2007) Perceptions of epigenetics. Nature 447:396–398PubMedCrossRefGoogle Scholar
  42. 42.
    Rhee I, Jair KW, Yen RW, Lengauer C, Herman JG, Kinzler KW, Vogelstein B, Baylin SB et al (2000) CpG methylation is maintained in human cancer cells lacking DNMT1. Nature 404:1003–1007PubMedCrossRefGoogle Scholar
  43. 43.
    Vertino PM, Yen RW, Gao J, Baylin SB (1996) De novo methylation of CpG island sequences in human fibroblasts overexpressing DNA (cytosine-5-)-methyltransferase. Mol Cell Biol 16:4555–4565PubMedPubMedCentralCrossRefGoogle Scholar
  44. 44.
    Bird AP, Wolffe AP (1999) Methylation-induced repression—belts, braces, and chromatin. Cell 99:451–454PubMedCrossRefGoogle Scholar
  45. 45.
    Jones PL, Veenstra GJ, Wade PA, Vermaak D, Kass SU, Landsberger N, Strouboulis J, Wolffe AP (1998) Methylated DNA and MeCP2 recruit histone deacetylase to repress transcription. Nat Genet 19:187–191PubMedCrossRefGoogle Scholar
  46. 46.
    Nan X, Ng HH, Johnson CA, Laherty CD, Turner BM, Eisenman RN, Bird A (1998) Transcriptional repression by the methyl-CpG-binding protein MeCP2 involves a histone deacetylase complex. Nature 393:386–389PubMedCrossRefGoogle Scholar
  47. 47.
    Ng HH, Zhang Y, Hendrich B, Johnson CA, Turner BM, Erdjument-Bromage H, Tempst P et al (1999) MBD2 is a transcriptional repressor belonging to the MeCP1 histone deacetylase complex. Nat Genet 23:58–61PubMedGoogle Scholar
  48. 48.
    Sarraf SA, Stancheva I (2004) Methyl-CpG binding protein MBD1 couples histone H3 methylation at lysine 9 by SETDB1 to DNA replication and chromatin assembly. Mol Cell 15:595–605PubMedCrossRefGoogle Scholar
  49. 49.
    Hashimoto H, Liu Y, Upadhyay AK, Chang Y, Howerton SB, Vertino PM, Zhang X, Cheng X (2012) Recognition and potential mechanisms for replication and erasure of cytosine hydroxymethylation. Nucleic Acids Res 40:4841–4849PubMedPubMedCentralCrossRefGoogle Scholar
  50. 50.
    Valinluck V, Sowers LC (2007) Endogenous cytosine damage products alter the site selectivity of human DNA maintenance methyltransferase DNMT1. Cancer Res 67:946–950PubMedCrossRefGoogle Scholar
  51. 51.
    He Y-F, Li B-Z, Li Z, Liu P, Wang Y, Tang Q, Ding J, Jia Y et al (2011) Tet-mediated formation of 5-carboxylcytosine and its excision by TDG in mammalian DNA. Science 333:1303–1307PubMedPubMedCentralCrossRefGoogle Scholar
  52. 52.
    Ito S, Shen L, Dai Q, Wu SC, Collins LB, Swenberg JA, He C, Zhang Y (2011) Tet proteins can convert 5-methylcytosine to 5-formylcytosine and 5-carboxylcytosine. Science 333:1300–1303PubMedPubMedCentralCrossRefGoogle Scholar
  53. 53.
    Tahiliani M, Koh KP, Shen Y, Pastor WA, Bandukwala H, Brudno Y, Agarwal S, Iyer LM et al (2009) Conversion of 5-methylcytosine to 5-hydroxymethylcytosine in mammalian DNA by MLL partner TET1. Science 324:930–935PubMedPubMedCentralCrossRefGoogle Scholar
  54. 54.
    Frauer C, Hoffmann T, Bultmann S, Casa V, Cardoso MC, Antes I, Leonhardt H (2011) Recognition of 5-hydroxymethylcytosine by the Uhrf1 SRA domain. PLoS One 6:e21306PubMedPubMedCentralCrossRefGoogle Scholar
  55. 55.
    Mellén M, Ayata P, Dewell S, Kriaucionis S, Heintz N (2012) MeCP2 binds to 5hmC enriched within active genes and accessible chromatin in the nervous system. Cell 151:1417–1430PubMedPubMedCentralCrossRefGoogle Scholar
  56. 56.
    Spruijt CG, Gnerlich F, Smits AH, Pfaffeneder T, Jansen PWTC, Bauer C, Münzel M, Wagner M et al (2013) Dynamic readers for 5-(hydroxy)methylcytosine and its oxidized derivatives. Cell 152:1146–1159PubMedCrossRefGoogle Scholar
  57. 57.
    Cech TR, Steitz JA (2014) The noncoding RNA revolution-trashing old rules to forge new ones. Cell 157:77–94PubMedCrossRefGoogle Scholar
  58. 58.
    Kouzarides T (2007) Chromatin modifications and their function. Cell 128:693–705PubMedCrossRefGoogle Scholar
  59. 59.
    Tammen SA, Friso S, Choi S-W (2012) Epigenetics: the link between nature and nurture. Mol Asp Med 34:753–764CrossRefGoogle Scholar
  60. 60.
    Thomsen R, Sølvsten CAE, Linnet TE, Blechingberg J, Nielsen AL (2010) Analysis of qPCR data by converting exponentially related Ct values into linearly related X0 values. J Bioinforma Comput Biol 8:885–900CrossRefGoogle Scholar
  61. 61.
    Shen L, Guo Y, Chen X, Ahmed S, Issa J-PJ (2007) Optimizing annealing temperature overcomes bias in bisulfite PCR methylation analysis. Biotechniques 42:48–50, 52 passimGoogle Scholar
  62. 62.
    Booth MJ, Branco MR, Ficz G, Oxley D, Krueger F, Reik W, Balasubramanian S (2012) Quantitative sequencing of 5-methylcytosine and 5-hydroxymethylcytosine at single-base resolution. Science 336:934–937PubMedCrossRefGoogle Scholar
  63. 63.
    Andersen CL, Jensen JL, Ørntoft TF (2004) Normalization of real-time quantitative reverse transcription-PCR data: a model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets. Cancer Res 64:5245–5250PubMedCrossRefGoogle Scholar
  64. 64.
    Adlard PA, Perreau VM, Engesser-Cesar C, Cotman CW (2004) The timecourse of induction of brain-derived neurotrophic factor mRNA and protein in the rat hippocampus following voluntary exercise. Neurosci Lett 363:43–48PubMedCrossRefGoogle Scholar
  65. 65.
    Baj G, D’Alessandro V, Musazzi L, Mallei A, Sartori CR, Sciancalepore M, Tardito D, Langone F et al (2012) Physical exercise and antidepressants enhance BDNF targeting in hippocampal CA3 dendrites: further evidence of a spatial code for BDNF splice variants. Neuropsychopharmacology 37:1–12CrossRefGoogle Scholar
  66. 66.
    Ieraci A, Mallei A, Musazzi L, Popoli M (2015) Physical exercise and acute restraint stress differentially modulate hippocampal brain-derived neurotrophic factor transcripts and epigenetic mechanisms in mice. Hippocampus 25:1380PubMedCrossRefGoogle Scholar
  67. 67.
    Ke Z, Yip SP, Li L, Zheng X-X, Tong K-Y (2011) The effects of voluntary, involuntary, and forced exercises on brain-derived neurotrophic factor and motor function recovery: a rat brain ischemia model. PLoS One 6:e16643PubMedPubMedCentralCrossRefGoogle Scholar
  68. 68.
    Neeper SA, Gómez-Pinilla F, Choi J, Cotman CW (1996) Physical activity increases mRNA for brain-derived neurotrophic factor and nerve growth factor in rat brain. Brain Res 726:49–56PubMedCrossRefGoogle Scholar
  69. 69.
    Oliff HS, Berchtold NC, Isackson P, Cotman CW (1998) Exercise-induced regulation of brain-derived neurotrophic factor (BDNF) transcripts in the rat hippocampus. Brain Res Mol Brain Res 61:147–153PubMedCrossRefGoogle Scholar
  70. 70.
    Li L-C, Dahiya R (2002) MethPrimer: designing primers for methylation PCRs. Bioinformatics 18:1427–1431PubMedCrossRefGoogle Scholar
  71. 71.
    Levy AP, Levy NS, Wegner S, Goldberg MA (1995) Transcriptional regulation of the rat vascular endothelial growth factor Gene by hypoxia. J Biol Chem 270:13333–13340PubMedCrossRefGoogle Scholar
  72. 72.
    Jin S-G, Wu X, Li AX, Pfeifer GP (2011) Genomic mapping of 5-hydroxymethylcytosine in the human brain. Nucleic Acids Res 39:5015–5024PubMedPubMedCentralCrossRefGoogle Scholar
  73. 73.
    Khare T, Pai S, Koncevicius K, Pal M, Kriukiene E, Liutkeviciute Z, Irimia M, Jia P et al (2012) 5-hmC in the brain is abundant in synaptic genes and shows differences at the exon-intron boundary. Nat Struct Mol Biol 19:1037–1043PubMedPubMedCentralCrossRefGoogle Scholar
  74. 74.
    Kriaucionis S, Heintz N (2009) The nuclear DNA base 5-hydroxymethylcytosine is present in Purkinje neurons and the brain. Science 324:929–930PubMedPubMedCentralCrossRefGoogle Scholar
  75. 75.
    Münzel M, Globisch D, Brückl T, Wagner M, Welzmiller V, Michalakis S, Müller M, Biel M et al (2010) Quantification of the sixth DNA base hydroxymethylcytosine in the brain. Angew Chem Int Ed Engl 49:5375–5377PubMedCrossRefGoogle Scholar
  76. 76.
    Creyghton MP, Cheng AW, Welstead GG, Kooistra T, Carey BW, Steine EJ, Hanna J, Lodato MA et al (2010) Histone H3K27ac separates active from poised enhancers and predicts developmental state. Proc Natl Acad Sci U S A 107:21931–21936PubMedPubMedCentralCrossRefGoogle Scholar
  77. 77.
    Pugh CW, Tan CC, Jones RW, Ratcliffe PJ (1991) Functional analysis of an oxygen-regulated transcriptional enhancer lying 3′ to the mouse erythropoietin gene. Proc Natl Acad Sci U S A 88:10553PubMedPubMedCentralCrossRefGoogle Scholar
  78. 78.
    Semenza GL, Nejfelt MK, Chi SM, Antonarakis SE (1991) Hypoxia-inducible nuclear factors bind to an enhancer element located 3′ to the human erythropoietin gene. Proc Natl Acad Sci U S A 88:5680–5684PubMedPubMedCentralCrossRefGoogle Scholar
  79. 79.
    Bechara RG, Lyne R, Kelly ÁM (2014) BDNF-stimulated intracellular signalling mechanisms underlie exercise-induced improvement in spatial memory in the male wistar rat. Behav Brain Res 275:297–306PubMedCrossRefGoogle Scholar
  80. 80.
    Lovatel GA, Elsner VR, Bertoldi K, Vanzella C, Moysés FDS, Vizuete A, Spindler C, Cechinel LR et al (2013) Treadmill exercise induces age-related changes in aversive memory, neuroinflammatory and epigenetic processes in the rat hippocampus. Neurobiol Learn Mem 101:94–102PubMedCrossRefGoogle Scholar
  81. 81.
    Uysal N, Kiray M, Sisman AR, Camsari UM, Gencoglu C, Baykara B, Cetinkaya C, Aksu I (2015) Effects of voluntary and involuntary exercise on cognitive functions, and VEGF and BDNF levels in adolescent rats. Biotech Histochem 90:55–68PubMedCrossRefGoogle Scholar
  82. 82.
    Alomari MA, Khabour OF, Alzoubi KH, Alzubi MA (2013) Forced and voluntary exercises equally improve spatial learning and memory and hippocampal BDNF levels. Behav Brain Res 247:34–39PubMedCrossRefGoogle Scholar
  83. 83.
    Ploughman M, Granter-Button S, Chernenko G, Tucker BA, Mearow KM, Corbett D (2005) Endurance exercise regimens induce differential effects on brain-derived neurotrophic factor, synapsin-I and insulin-like growth factor I after focal ischemia. Neuroscience 136:991–1001PubMedCrossRefGoogle Scholar
  84. 84.
    van Praag H, Shubert T, Zhao C, Gage FH (2005) Exercise enhances learning and hippocampal neurogenesis in aged mice. J Neurosci 25:8680–8685PubMedPubMedCentralCrossRefGoogle Scholar
  85. 85.
    Yau S, Gil-Mohapel J, Christie BR, So K (2014) Physical exercise-induced adult neurogenesis: a good strategy to prevent cognitive decline in neurodegenerative diseases? Biomed Res Int 2014:403120PubMedPubMedCentralCrossRefGoogle Scholar
  86. 86.
    Kuhn HG, Dickinson-Anson H, Gage FH (1996) Neurogenesis in the dentate gyrus of the adult rat: age-related decrease of neuronal progenitor proliferation. J Neurosci 16:2027–2033PubMedGoogle Scholar
  87. 87.
    Luskin MB, Boone MS (1994) Rate and pattern of migration of lineally-related olfactory bulb interneurons generated postnatally in the subventricular zone of the rat. Chem Senses 19:695–714PubMedCrossRefGoogle Scholar
  88. 88.
    Cao L, Jiao X, Zuzga DS, Liu Y, Fong DM, Young D, During MJ (2004) VEGF links hippocampal activity with neurogenesis, learning and memory. Nat Genet 36:827–835PubMedCrossRefGoogle Scholar
  89. 89.
    van Praag H, Christie BR, Sejnowski TJ, Gage FH (1999a) Running enhances neurogenesis, learning, and long-term potentiation in mice. Proc Natl Acad Sci U S A 96:13427–13431PubMedPubMedCentralCrossRefGoogle Scholar
  90. 90.
    Black JE, Isaacs KR, Anderson BJ, Alcantara AA, Greenough WT (1990) Learning causes synaptogenesis, whereas motor activity causes angiogenesis, in cerebellar cortex of adult rats. Proc Natl Acad Sci U S A 87:5568–5572PubMedPubMedCentralCrossRefGoogle Scholar
  91. 91.
    Kleim JA, Cooper NR, Vanden Berg PM (2002) Exercise induces angiogenesis but does not alter movement representations within rat motor cortex. Brain Res 934:1–6PubMedCrossRefGoogle Scholar
  92. 92.
    Swain RA, Harris AB, Wiener EC, Dutka MV, Morris HD, Theien BE, Konda S, Engberg K et al (2003) Prolonged exercise induces angiogenesis and increases cerebral blood volume in primary motor cortex of the rat. Neuroscience 117:1037–1046PubMedCrossRefGoogle Scholar
  93. 93.
    Palmer TD, Willhoite AR, Gage FH (2000) Vascular niche for adult hippocampal neurogenesis. J Comp Neurol 425:479–494PubMedCrossRefGoogle Scholar
  94. 94.
    Pereira AC, Huddleston DE, Brickman AM, Sosunov AA, Hen R, McKhann GM, Sloan R, Gage FH et al (2007) An in vivo correlate of exercise-induced neurogenesis in the adult dentate gyrus. Proc Natl Acad Sci U S A 104:5638–5643PubMedPubMedCentralCrossRefGoogle Scholar
  95. 95.
    Ntanasis-Stathopoulos J, Tzanninis JG, Philippou A, Koutsilieris M (2013) Epigenetic regulation on gene expression induced by physical exercise. J Musculoskelet Neuronal Interact 13:133–146PubMedGoogle Scholar
  96. 96.
    Spindler C, Cechinel LR, Basso C, Moysés F, Bertoldi K, Roesler R, Lovatel GA, Rostirola Elsner V et al (2014) Treadmill exercise alters histone acetyltransferases and histone deacetylases activities in frontal cortices from wistar rats. Cell Mol Neurobiol 34:1097–1101PubMedCrossRefGoogle Scholar
  97. 97.
    Hahn MA, Szabó PE, Pfeifer GP (2014) 5-hydroxymethylcytosine: a stable or transient DNA modification? Genomics 104:314–323PubMedPubMedCentralCrossRefGoogle Scholar
  98. 98.
    Douet V, Heller MB, Le Saux O (2007) DNA methylation and Sp1 binding determine the tissue-specific transcriptional activity of the mouse Abcc6 promoter. Biochem Biophys Res Commun 354:66–71PubMedCrossRefGoogle Scholar
  99. 99.
    Tian H-P, Lun S-M, Huang H-J, He R, Kong P-Z, Wang Q-S, Li X-Q, Feng Y-M (2015) DNA methylation affects the SP1-regulated transcription of FOXF2 in breast cancer cells. J Biol Chem 290:19173–19183PubMedPubMedCentralCrossRefGoogle Scholar
  100. 100.
    Déry M-AC, Michaud MD, Richard DE (2005) Hypoxia-inducible factor 1: regulation by hypoxic and non-hypoxic activators. Int J Biochem Cell Biol 37:535–540PubMedCrossRefGoogle Scholar
  101. 101.
    Mistry IN, Smith PJS, Wilson DI, Tavassoli A (2015) Probing the epigenetic regulation of HIF-1α transcription in developing tissue. Mol BioSyst 11:2780–2785PubMedCrossRefGoogle Scholar
  102. 102.
    Wenger RH, Kvietikova I, Rolfs A, Camenisch G, Gassmann M (1998) Oxygen-regulated erythropoietin gene expression is dependent on a CpG methylation-free hypoxia-inducible factor-1 DNA-binding site. Eur J Biochem 253:771–777PubMedCrossRefGoogle Scholar
  103. 103.
    Fish JE, Yan MS, Matouk CC, St Bernard R, Ho JJD, Gavryushova A, Srivastava D, Marsden PA (2010) Hypoxic repression of endothelial nitric-oxide synthase transcription is coupled with eviction of promoter histones. J Biol Chem 285:810–826PubMedCrossRefGoogle Scholar
  104. 104.
    Fu S-P, He S-Y, Xu B, Hu C-J, Lu S-F, Shen W-X, Huang Y, Hong H et al (2014) Acupuncture promotes angiogenesis after myocardial ischemia through H3K9 acetylation regulation at VEGF gene. PLoS One 9:e94604PubMedPubMedCentralCrossRefGoogle Scholar
  105. 105.
    Johnson AB, Denko N, Barton MC (2008) Hypoxia induces a novel signature of chromatin modifications and global repression of transcription. Mutat Res 640:174–179PubMedPubMedCentralCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Department of BiomedicineAarhus UniversityAarhus CDenmark
  2. 2.The Lundbeck Foundation Initiative for Integrative Psychiatric Research, iPSYCHAarhus CDenmark
  3. 3.Centre for Integrative Sequencing, iSEQAarhus UniversityAarhus CDenmark

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