Journal of Molecular Neuroscience

, Volume 61, Issue 3, pp 436–448 | Cite as

Estrogen Modulates ubc9 Expression and Synaptic Redistribution in the Brain of APP/PS1 Mice and Cortical Neurons

  • Yu-Jie Lai
  • Lu Liu
  • Xiao-Tong Hu
  • Ling He
  • Guo-Jun ChenEmail author


Estrogen exerts multiple actions in the brain and is an important neuroprotective factor in a number of neuronal disorders. However, the underlying mechanism remains unknown. Studies demonstrate that ubiquitin-conjugating enzyme 9 (ubc9) has an integral role in synaptic plasticity and may contribute to the pathology of neuronal disorders. We aimed to investigate the effects of estrogen on ubc9 and in the Alzheimer’s disease brain. Ubc9 protein and mRNA were significantly increased in the cortex and hippocampus of APP/PS1 mice with enhanced SUMOylation. Systemic estrogen administration led to reduced ubc9 expression in ovariectomized APP/PS1 mice and reduced SUMOylation. The inhibition of ubc9 expression by estrogen was found to be dose-dependent in cultured neurons. However, estrogen receptor (ER) antagonist ICI182780 did not block the inhibition of ubc9 expression by estrogen. Furthermore, the reduced expression of ubc9 was not mediated by ERα or ERβ agonists alone or in combination, but by the membrane-impermeable ER agonist E2-bovine serum albumin. The activation of the G protein-coupled ER mediated the inhibition of ubc9 expression of estrogen. A phosphoinositide 3-kinase (PI3K) inhibitor, rather than an extracellular signal-regulated kinase inhibitor, blocked the inhibition of ubc9 by estrogen. Estrogen treatment significantly increased the phosphorylation of PI3K, which suggests that activation of the PI3K pathway by estrogen is required for ubc9 regulation. Further, ubc9 interacted with the synaptic proteins post-synaptic density protein 95 (PSD95) and synaptophysin. Estrogen decreased the interaction of ubc9 with post-synaptic PSD95, but increased the interaction of ubc9 with pre-synaptic synaptophysin. These results suggest that a membrane-bound ER might mediate the estrogen inhibition of ubc9 in cortical neurons, where PI3K plays an important role. We also show that ubc9 can interact with synaptic proteins, which are subject to estrogen regulation.


Estrogen ubc9 Neuroplasticity PSD95 Synaptophysin 



This work was supported by NSFC grants (numbers 81171197 and 81220108010) and a Bureau of Health of Chongqing Medical Research grant (number 2011-1-018) to G-J. C.

Compliance with Ethical Standards

All protocols were approved by the Commission of Chongqing Medical University for Ethics of Experiments on Animals and were conducted in accordance with international standards.

Competing Interests

The authors declare that they have no competing interests.

Supplementary material

12031_2017_884_Fig1_ESM.gif (110 kb)

Supplementary figure legend. Estrogen decreases ubc9 expression in cultured cortical neurons. Primary cultured cortical neurons were treated with vehicle (DMSO), 17β-estradiol, the non-selective estrogen antagonist ICI, or 17β-estradiol and ICI. (A) Estrogen dose-dependently decreased ubc9 expression in cortical neurons. Neurons were incubated with vehicle DMSO or various 17β-estradiol (E2) concentrations (1 nM, 10 nM, 100 nM or 1 μM) for 24 h, and ubc9 protein was then measured, which were performed in triplicate. (B) Estrogen time-dependently decreased ubc9 expression in cortical neurons. Time-course response showed that ubc9 protein levels were decreased within 24 h, and remained to be up to 72 h in the presence of E2 (10 nM, Fig. B),which were performed in triplicate. (C) ICI182780 failed to block the effects of estrogen on ubc9 expression. We pre-incubated the neurons with the non-selective ER antagonist ICI182780 at a range of concentrations (1 μM, 10 μM, and 100 μM), which were performed in triplicate. (GIF 109 kb)

12031_2017_884_MOESM1_ESM.tif (1.7 mb)
High Resolution Image (TIFF 1732 kb)


  1. Bernelot Moens SJ et al (2012) Rapid estrogen receptor signaling is essential for the protective effects of estrogen against vascular injury. Circulation 126:1993–2004. doi: 10.1161/circulationaha.112.124529 CrossRefPubMedPubMedCentralGoogle Scholar
  2. Bi R, Foy MR, Vouimba RM, Thompson RF, Baudry M (2001) Cyclic changes in estradiol regulate synaptic plasticity through the MAP kinase pathway. Proc Natl Acad Sci U S A 98:13391–13395. doi: 10.1073/pnas.241507698 CrossRefPubMedPubMedCentralGoogle Scholar
  3. Briz V, Baudry M (2014) Estrogen regulates protein synthesis and actin polymerization in hippocampal neurons through different molecular mechanisms. Front Endocrinol 5:22. doi: 10.3389/fendo.2014.00022 CrossRefGoogle Scholar
  4. Choi JH, Park JY, Park SP, Lee H, Han S, Park KH, Suh YH (2016) Regulation of mGluR7 trafficking by SUMOylation in neurons. Neuropharmacology 102:229–235. doi: 10.1016/j.neuropharm.2015.11.021 CrossRefPubMedGoogle Scholar
  5. Christensen A, Micevych P (2013) A novel membrane estrogen receptor activated by STX induces female sexual receptivity through an interaction with mGluR1a. Neuroendocrinology 97:363–368. doi: 10.1159/000351077 CrossRefPubMedPubMedCentralGoogle Scholar
  6. Chu Z, Andrade J, Shupnik MA, Moenter SM (2009) Differential regulation of gonadotropin-releasing hormone neuron activity and membrane properties by acutely applied estradiol: dependence on dose and estrogen receptor subtype. J Neurosci : Off JSoc Neurosci 29:5616–5627. doi: 10.1523/jneurosci.0352-09.2009 CrossRefGoogle Scholar
  7. DeVoogd T, Nottebohm F (1981) Gonadal hormones induce dendritic growth in the adult avian brain. Science 214:202–204CrossRefPubMedGoogle Scholar
  8. Ding D et al (2014) Therapeutic implications of estrogen for cerebral vasospasm and delayed cerebral ischemia induced by aneurysmal subarachnoid hemorrhage. Biomed Res Int 2014:727428. doi: 10.1155/2014/727428 PubMedPubMedCentralGoogle Scholar
  9. Ehlers MD (2003) Activity level controls postsynaptic composition and signaling via the ubiquitin-proteasome system. Nat Neurosci 6:231–242. doi: 10.1038/nn1013 CrossRefPubMedGoogle Scholar
  10. Fan X, Jin WY, Lu J, Wang J, Wang YT (2014) Rapid and reversible knockdown of endogenous proteins by peptide-directed lysosomal degradation. Nat Neurosci 17:471–480. doi: 10.1038/nn.3637 CrossRefPubMedPubMedCentralGoogle Scholar
  11. Feligioni M, Nishimune A, Henley JM (2009) Protein SUMOylation modulates calcium influx and glutamate release from presynaptic terminals. Eur J Neurosci 29:1348–1356. doi: 10.1111/j.1460-9568.2009.06692.x CrossRefPubMedPubMedCentralGoogle Scholar
  12. Geerlings MI et al (2003) Endogenous estradiol and risk of dementia in women and men: the Rotterdam study. Ann Neurol 53:607–615. doi: 10.1002/ana.10521 CrossRefPubMedGoogle Scholar
  13. Gjoneska E, Pfenning AR, Mathys H, Quon G, Kundaje A, Tsai LH, Kellis M (2015) Conserved epigenomic signals in mice and humans reveal immune basis of Alzheimer’s disease. Nature 518:365–369. doi: 10.1038/nature14252 CrossRefPubMedPubMedCentralGoogle Scholar
  14. Grove-Strawser D, Boulware MI, Mermelstein PG (2010) Membrane estrogen receptors activate the metabotropic glutamate receptors mGluR5 and mGluR3 to bidirectionally regulate CREB phosphorylation in female rat striatal neurons. Neuroscience 170:1045–1055. doi: 10.1016/j.neuroscience.2010.08.012 CrossRefPubMedPubMedCentralGoogle Scholar
  15. Henley JM, Craig TJ, Wilkinson KA (2014) Neuronal SUMOylation: mechanisms, physiology, and roles in neuronal dysfunction. Physiol Rev 94:1249–1285. doi: 10.1152/physrev.00008.2014 CrossRefPubMedPubMedCentralGoogle Scholar
  16. Hogervorst E, Williams J, Combrinck M, David Smith A (2003) Measuring serum oestradiol in women with Alzheimer’s disease: the importance of the sensitivity of the assay method. Eur J Endocrinol/Eur Fed Endocr Soc 148:67–72CrossRefGoogle Scholar
  17. Hoppe JB, Salbego CG, Cimarosti H (2015) SUMOylation: novel neuroprotective approach for Alzheimer’s disease? Aging Dis 6:322–330. doi: 10.14336/ad.2014.1205 CrossRefPubMedPubMedCentralGoogle Scholar
  18. Kessels HW, Malinow R (2009) Synaptic AMPA receptor plasticity and behavior. Neuron 61:340–350. doi: 10.1016/j.neuron.2009.01.015 CrossRefPubMedPubMedCentralGoogle Scholar
  19. Lee L et al (2014) Regulation of synaptic plasticity and cognition by SUMO in normal physiology and Alzheimer’s disease. Sci Rep 4:7190. doi: 10.1038/srep07190 CrossRefPubMedPubMedCentralGoogle Scholar
  20. Lee L et al (2015) Corrigendum: regulation of synaptic plasticity and cognition by SUMO in normal physiology and Alzheimer’s disease. Sci Rep 5:11782. doi: 10.1038/srep11782 CrossRefPubMedPubMedCentralGoogle Scholar
  21. Liu L, Zhao Y, Xie K, Sun X, Gao Y, Wang Z (2013) Estrogen-induced nongenomic calcium signaling inhibits lipopolysaccharide-stimulated tumor necrosis factor alpha production in macrophages. PLoS One 8:e83072. doi: 10.1371/journal.pone.0083072 CrossRefPubMedPubMedCentralGoogle Scholar
  22. Liu SB et al (2012) G-protein-coupled receptor 30 mediates rapid neuroprotective effects of estrogen via depression of NR2B-containing NMDA receptors. J Neurosci : Off J Soc Neurosci 32:4887–4900. doi: 10.1523/jneurosci.5828-11.2012 CrossRefGoogle Scholar
  23. Loriol C, Khayachi A, Poupon G, Gwizdek C, Martin S (2013) Activity-dependent regulation of the sumoylation machinery in rat hippocampal neurons biology of the cell / under the auspices of the European Cell Biology Organization 105:30–45 doi: 10.1111/boc.201200016
  24. Loriol C, Parisot J, Poupon G, Gwizdek C, Martin S (2012) Developmental regulation and spatiotemporal redistribution of the sumoylation machinery in the rat central nervous system. PLoS One 7:e33757. doi: 10.1371/journal.pone.0033757 CrossRefPubMedPubMedCentralGoogle Scholar
  25. Luo HB et al (2014) SUMOylation at K340 inhibits tau degradation through deregulating its phosphorylation and ubiquitination. Proc Natl Acad Sci U S A 111:16586–16591. doi: 10.1073/pnas.1417548111 CrossRefPubMedPubMedCentralGoogle Scholar
  26. Mannella P, Brinton RD (2006) Estrogen receptor protein interaction with phosphatidylinositol 3-kinase leads to activation of phosphorylated Akt and extracellular signal-regulated kinase 1/2 in the same population of cortical neurons: a unified mechanism of estrogen action. J Neurosci: Off J Soc Neurosci 26:9439–9447. doi: 10.1523/jneurosci.1443-06.2006 CrossRefGoogle Scholar
  27. Martin S, Nishimune A, Mellor JR, Henley JM (2007a) SUMOylation regulates kainate-receptor-mediated synaptic transmission. Nature 447:321–325. doi: 10.1038/nature05736 CrossRefPubMedPubMedCentralGoogle Scholar
  28. Martin S, Wilkinson KA, Nishimune A, Henley JM (2007b) Emerging extranuclear roles of protein SUMOylation in neuronal function and dysfunction. Nat Rev Neurosci 8:948–959. doi: 10.1038/nrn2276 CrossRefPubMedPubMedCentralGoogle Scholar
  29. Martins WC, Tasca CI, Cimarosti H (2015) Battling Alzheimer’s disease: targeting SUMOylation-mediated pathways. Neurochem Res. doi: 10.1007/s11064-015-1681-3 Google Scholar
  30. McEwen BS, Alves SE (1999) Estrogen actions in the central nervous system. Endocr Rev 20:279–307. doi: 10.1210/edrv.20.3.0365 PubMedGoogle Scholar
  31. McMillan LE, Brown JT, Henley JM, Cimarosti H (2011) Profiles of SUMO and ubiquitin conjugation in an Alzheimer’s disease model. Neurosci Lett 502:201–208. doi: 10.1016/j.neulet.2011.07.045 CrossRefPubMedPubMedCentralGoogle Scholar
  32. Murdock DG et al (2013) KIAA1462, a coronary artery disease associated gene, is a candidate gene for late onset Alzheimer disease in APOE carriers. PLoS One 8:e82194. doi: 10.1371/journal.pone.0082194 CrossRefPubMedPubMedCentralGoogle Scholar
  33. Murray CJ et al (2013) The state of US health, 1990-2010: burden of diseases, injuries, and risk factors. JAMA 310:591–608. doi: 10.1001/jama.2013.13805 CrossRefPubMedGoogle Scholar
  34. Nistico R, Ferraina C, Marconi V, Blandini F, Negri L, Egebjerg J, Feligioni M (2014) Age-related changes of protein SUMOylation balance in the AbetaPP Tg2576 mouse model of Alzheimer’s disease. Front Pharmacol 5:63. doi: 10.3389/fphar.2014.00063 PubMedPubMedCentralGoogle Scholar
  35. Okun E, Arumugam TV, Tang SC, Gleichmann M, Albeck M, Sredni B, Mattson MP (2007) The organotellurium compound ammonium trichloro(dioxoethylene-0,0′) tellurate enhances neuronal survival and improves functional outcome in an ischemic stroke model in mice. J Neurochem 102:1232–1241. doi: 10.1111/j.1471-4159.2007.04615.x CrossRefPubMedGoogle Scholar
  36. Olzscha H et al (2011) Amyloid-like aggregates sequester numerous metastable proteins with essential cellular functions. Cell 144:67–78. doi: 10.1016/j.cell.2010.11.050 CrossRefPubMedGoogle Scholar
  37. Ordonez-Moran P, Munoz A (2009) Nuclear receptors: genomic and non-genomic effects converge. Cell Cycle (Georgetown, Tex) 8:1675–1680. doi: 10.4161/cc.8.11.8579 CrossRefGoogle Scholar
  38. Paul SM, Purdy RH (1992) Neuroactive steroids. FASEB J: Off Publ Fed Am Soc Exp Biol 6:2311–2322Google Scholar
  39. Peineau S et al (2007) LTP inhibits LTD in the hippocampus via regulation of GSK3beta. Neuron 53:703–717. doi: 10.1016/j.neuron.2007.01.029 CrossRefPubMedGoogle Scholar
  40. Qiu J, Bosch MA, Tobias SC, Grandy DK, Scanlan TS, Ronnekleiv OK, Kelly MJ (2003) Rapid signaling of estrogen in hypothalamic neurons involves a novel G-protein-coupled estrogen receptor that activates protein kinase C. J Neurosci: J Soc Neurosci 23:9529–9540Google Scholar
  41. Revankar CM, Cimino DF, Sklar LA, Arterburn JB, Prossnitz ER (2005) A transmembrane intracellular estrogen receptor mediates rapid cell signaling. Science (New York, NY) 307:1625–1630. doi: 10.1126/science.1106943 CrossRefGoogle Scholar
  42. Sa SI, Fonseca BM, Teixeira N, Madeira MD (2015) Estrogen receptors alpha and beta have different roles in the induction and trafficking of progesterone receptors in hypothalamic ventromedial neurons. FEBS J 282:1126–1136. doi: 10.1111/febs.13207 CrossRefPubMedGoogle Scholar
  43. Sanden C et al (2011) G protein-coupled estrogen receptor 1/G protein-coupled receptor 30 localizes in the plasma membrane and traffics intracellularly on cytokeratin intermediate filaments. Mol Pharmacol 79:400–410. doi: 10.1124/mol.110.069500 CrossRefPubMedGoogle Scholar
  44. Sarkar SN, Huang RQ, Logan SM, Yi KD, Dillon GH, Simpkins JW (2008) Estrogens directly potentiate neuronal L-type Ca2+ channels. Proc Natl Acad Sci U S A 105:15148–15153. doi: 10.1073/pnas.0802379105 CrossRefPubMedPubMedCentralGoogle Scholar
  45. Scheschonka A, Tang Z, Betz H (2007) Sumoylation in neurons: nuclear and synaptic roles? Trends Neurosci 30:85–91. doi: 10.1016/j.tins.2007.01.003 CrossRefPubMedGoogle Scholar
  46. Schonknecht P, Henze M, Hunt A, Klinga K, Haberkorn U, Schroder J (2003) Hippocampal glucose metabolism is associated with cerebrospinal fluid estrogen levels in postmenopausal women with Alzheimer’s disease. Psychiatry Res 124:125–127CrossRefPubMedGoogle Scholar
  47. Scudder SL, Goo MS, Cartier AE, Molteni A, Schwarz LA, Wright R, Patrick GN (2014) Synaptic strength is bidirectionally controlled by opposing activity-dependent regulation of Nedd4-1 and USP8. J Neurosci: Off J Soc Neurosci 34:16637–16649. doi: 10.1523/jneurosci.2452-14.2014 CrossRefGoogle Scholar
  48. Srivastava DP, Woolfrey KM, Penzes P (2013) Insights into rapid modulation of neuroplasticity by brain estrogens. Pharmacol Rev 65:1318–1350. doi: 10.1124/pr.111.005272 CrossRefPubMedPubMedCentralGoogle Scholar
  49. Tan Z, Zhou LJ, Li Y, Cui YH, Xiang QL, Lin GP, Wang TH (2012) E(2)-BSA activates caveolin-1 via PI(3)K/ERK1/2 and lysosomal degradation pathway and contributes to EPC proliferation. Int J Cardiol 158:46–53. doi: 10.1016/j.ijcard.2010.12.106 CrossRefPubMedGoogle Scholar
  50. Toran-Allerand CD et al (2002) ER-X: a novel, plasma membrane-associated, putative estrogen receptor that is regulated during development and after ischemic brain injury. J Neurosci: Off J Soc Neurosci 22:8391–8401Google Scholar
  51. Toran-Allerand CD, Miranda RC, Bentham WD, Sohrabji F, Brown TJ, Hochberg RB, NJ ML (1992) Estrogen receptors colocalize with low-affinity nerve growth factor receptors in cholinergic neurons of the basal forebrain. Proc Natl Acad Sci U S A 89:4668–4672CrossRefPubMedPubMedCentralGoogle Scholar
  52. Woolley CS, Gould E, Frankfurt M, McEwen BS (1990) Naturally occurring fluctuation in dendritic spine density on adult hippocampal pyramidal neurons. J Neurosci: Off J Soc Neurosci 10:4035–4039Google Scholar
  53. Wu J et al (2011) Arc/Arg3.1 regulates an endosomal pathway essential for activity-dependent beta-amyloid generation. Cell 147:615–628. doi: 10.1016/j.cell.2011.09.036 CrossRefPubMedPubMedCentralGoogle Scholar
  54. Yun SM et al (2013) SUMO1 modulates Abeta generation via BACE1 accumulation. Neurobiol Aging 34:650–662. doi: 10.1016/j.neurobiolaging.2012.08.005 CrossRefPubMedGoogle Scholar
  55. Zhang J et al (2011) The AAA+ ATPase Thorase regulates AMPA receptor-dependent synaptic plasticity and behavior. Cell 145:284–299. doi: 10.1016/j.cell.2011.03.016 CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Yu-Jie Lai
    • 1
  • Lu Liu
    • 1
  • Xiao-Tong Hu
    • 1
  • Ling He
    • 1
  • Guo-Jun Chen
    • 1
    Email author
  1. 1.Department of Neurology, Chongqing Key Laboratory of NeurologyHospital of Chongqing Medical UniversityChongqingChina

Personalised recommendations