Molecular Neurobiology

, Volume 55, Issue 5, pp 3856–3865 | Cite as

Weakened Intracellular Zn2+-Buffering in the Aged Dentate Gyrus and Its Involvement in Erasure of Maintained LTP

  • Atsushi Takeda
  • Haruna Tamano
  • Taku Murakami
  • Hiroyuki Nakada
  • Tatsuya Minamino
  • Yuta Koike


Memory is lost by the increased influx of extracellular Zn2+ into neurons. It is possible that intracellular Zn2+ dynamics is modified even at non-zincergic medial perforant pathway-dentate granule cell synapses along with aging and that vulnerability to the modification is linked to age-related cognitive decline. To examine these possibilities, vulnerability of long-term potentiation (LTP) maintenance, which underlies memory retention, to modification of synaptic Zn2+ dynamics was compared between young and aged rats. The influx of extracellular Zn2+ into dentate granule cells was increased in aged rats after injection of high K+ into the dentate gyrus, but not in young rats. This increase impaired maintained LTP in aged rats. However, the impairment was rescued by co-injection of CaEDTA, an extracellular Zn2+ chelator, or CNQX, an AMPA receptor antagonist, which suppressed the Zn2+ influx. Maintained LTP was also impaired in aged rats after injection of ZnAF-2DA into the dentate gyrus that chelates intracellular Zn2+, but not in young rats. Interestingly, the capacity of chelating intracellular Zn2+ with intracellular ZnAF-2 was almost lost in the aged dentate gyrus 2 h after injection of ZnAF-2DA into the dentate gyrus, suggesting that intracellular Zn2+-buffering is weakened in the aged dentate gyrus, compared to the young dentate gyrus. In the dentate gyrus of aged rats, maintained LTP is more vulnerable to modification of intracellular Zn2+ dynamics than in young rats, probably due to weakened intracellular Zn2+-buffering.


Zn2+-buffering Dentate granule cell LTP maintenance AMPA receptor Aging 


Compliance with Ethical Standards

All the experiments were performed in accordance with the Guidelines for the Care and Use of Laboratory Animals of the University of Shizuoka that refer to the American Association for Laboratory Animals Science and the guidelines laid down by the NIH (NIH Guide for the Care and Use of Laboratory Animals) in the USA. The Ethics Committee for Experimental Animals in the University of Shizuoka has approved this work (approval numbers: 136043)

Conflict of Interest

The authors declare that they have no conflict of interest.


  1. 1.
    Bliss TV, Collingridge GL (1993) A synaptic model of memory: long-term potentiation in the hippocampus. Nature 361:31–39CrossRefPubMedGoogle Scholar
  2. 2.
    Nicoll RA, Malenka RC (1995) Contrasting properties of two forms of long-term potentiation in the hippocampus. Nature 377:115–118CrossRefPubMedGoogle Scholar
  3. 3.
    Malenka RC, Nicoll RA (1999) Long-term potentiation-a decade of progress? Science 285:1870–1874CrossRefPubMedGoogle Scholar
  4. 4.
    Neves G, Cooke SF, Bliss TVP (2008) Synaptic plasticity, memory and the hippocampus: a neural network approach to causality. Nat Rev Neurosci 9:65–67CrossRefPubMedGoogle Scholar
  5. 5.
    Ceccom J, Halley H, Daumas S, Lassalle JM (2014) A specific role for hippocampal mossy fiber's zinc in rapid storage of emotional memories. Learn Mem 21:287–297CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Takeda A, Tamano H (2016) Significance of the degree of synaptic Zn2+ signaling in cognition. Biometals 29:177–185CrossRefPubMedGoogle Scholar
  7. 7.
    Sindreu CB, Varoqui H, Erickson JD, Pérez-Clausell J (2003) Boutons containing vesicular zinc define a subpopulation of synapses with low AMPAR content in rat hippocampus. Cereb Cortex 13:823–829CrossRefPubMedGoogle Scholar
  8. 8.
    Brown MW, Aggleton JP (2001) Recognition memory: what are the roles of the perirhinal cortex and hippocampus? Nat Rev Neurosci 2:51–61CrossRefPubMedGoogle Scholar
  9. 9.
    Malleret G, Alarcon JM, Martel G, Takizawa S, Vronskaya S, Yin D (2010) Bidirectional regulation of hippocampal long-term synaptic plasticity and its influence on opposing forms of memory. J Neurosci 30:3813–3825CrossRefPubMedGoogle Scholar
  10. 10.
    Knierim JJ, Neunuebel JP (2016) Tracking the flow of hippocampal computation: Pattern separation, pattern completion, and attractor dynamics. Neurobiol Learn Mem 129:38–49CrossRefPubMedGoogle Scholar
  11. 11.
    Takeda A, Tamano H, Ogawa T, Takada S, Nakamura M, Fujii H, Ando M (2014) Intracellular Zn2+ signaling in the dentate gyrus is required for object recognition memory. Hippocampus 24:1404–1412CrossRefPubMedGoogle Scholar
  12. 12.
    Pastalkova E, Serrano P, Pinkhasova D, Wallace E, Fenton AA, Sacktor TC (2006) Storage of spatial information by the maintenance mechanism of LTP. Science 313:1141–1144CrossRefPubMedGoogle Scholar
  13. 13.
    Tamano H, Minamino T, Fujii H, Takada S, Nakamura M, Ando M, Takeda A (2015) Blockade of intracellular Zn2+ signaling in the dentate gyrus erases recognition memory via impairment of maintained LTP. Hippocampus 25:952–962CrossRefPubMedGoogle Scholar
  14. 14.
    Takeda A, Tamano H (2016) Innervation from the entorhinal cortex to the dentate gyrus and the vulnerability to Zn2+. J Trace Elem Med Biol 38:19–23CrossRefPubMedGoogle Scholar
  15. 15.
    Lee JY, Kim JS, Byun HR, Palmiter RD, Koh JY (2011) Dependence of the histofluorescently reactive zinc pool on zinc transporter-3 in the normal brain. Brain Res 1418:12–22CrossRefPubMedGoogle Scholar
  16. 16.
    Saito T, Takahashi K, Nakagawa N, Hosokawa T, Kurasaki M, Yamanoshita O, Yamamoto Y, Sasaki H et al (2000) Biochem Biophys Res Commun 279:505–511CrossRefPubMedGoogle Scholar
  17. 17.
    Adlard PA, Parncutt JM, Finkelstein DI, Bush AI (2010) Cognitive loss in zinc transporter-3 knock-out mice: a phenocopy for the synaptic and memory deficits of Alzheimer's disease? J Neurosci 30:1631–1636CrossRefPubMedGoogle Scholar
  18. 18.
    Sindreu C, Palmiter RD, Storm DR (2011) Zinc transporter ZnT-3 regulates presynaptic Erk1/2 signaling and hippocampus-dependent memory. Proc Natl Acad Sci U S A 108:3366–3370CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Adlard PA, Sedjahtera A, Gunawan L, Bray L, Hare D, Lear J, Doble P, Bush AI et al (2014) A novel approach to rapidly prevent age-related cognitive decline. Aging Cell 13:351–359CrossRefPubMedGoogle Scholar
  20. 20.
    Adlard PA, Parncutt J, Lal V, James S, Hare D, Doble P, Finkelstein DI, Bush AI (2015) Metal chaperones prevent zinc-mediated cognitive decline. Neurobiol Dis 81:196–202CrossRefPubMedGoogle Scholar
  21. 21.
    Takeda A, Koike Y, Osaw M, Tamano H (2017) Characteristic of extracellular Zn2+ influx in the middle-aged dentate gyrus and its involvement in attenuation of LTP. Mol Neurobiol. doi: 10.1007/s12035-017-0472-z
  22. 22.
    Suzuki M, Fujise Y, Tsuchiya Y, Tamano H, Takeda A (2015) Excess influx of Zn2+ into dentate granule cells affects object recognition memory via attenuated LTP. Neurochem Int 87:60–65CrossRefPubMedGoogle Scholar
  23. 23.
    Takeda A, Tamano H, Hisatsune M, Murakami T, Nakada H, Fujii H (2017) Maintained LTP and memory are lost by Zn2+ influx into dentate granule cells, but not Ca2+ influx. Mol Neurobiol. doi: 10.1007/s12035-017-0428-3
  24. 24.
    Hirano T, Kikuchi K, Urano Y, Nagano T (2002) Improvement and biological applications of fluorescent probes for zinc, ZnAFs. J Am Chem Soc 124:6555–6562CrossRefPubMedGoogle Scholar
  25. 25.
    Ueno S, Tsukamoto M, Hirano T, Kikuchi K, Yamada MK, Nishiyama N, Nagano T, Matsuki N et al (2002) Mossy fiber Zn2+ spillover modulates heterosynaptic N-methyl-D-aspartate receptor activity in hippocampal CA3 circuits. J Cell Biol 158:215–220CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Fukazawa Y, Saitoh Y, Ozawa F, Ohta Y, Mizuno K, Inokuchi K (2003) Hippocampal LTP is accompanied by enhanced F-actin content within the dendritic spine that is essential for late LTP maintenance in vivo. Neuron 38:447–460CrossRefPubMedGoogle Scholar
  27. 27.
    McNaughton BL, Barnes CA (1997) Physiological identification and analysis of dentate granule cell responses to stimulation of the medial and lateral perforant pathways in the rat. J Comp Neurol 175:439–454CrossRefGoogle Scholar
  28. 28.
    Takeda A, Nakamura M, Fujii H, Uematsu C, Minamino T, Adlard PA, Bush AI, Tamano H (2014) Amyloid β-mediated Zn2+ influx into dentate granule cells transiently induces a short-term cognitive deficit. PLoS One 9:e115923CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Small SA, Schobel SA, Buxton RB, Witter MP, Barnes CA (2011) A pathophysiological framework of hippocampal dysfunction in ageing and disease. Nat Rev Neurosci 12:585–601CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Brickman AM, Khan UA, Provenzano FA, Yeung LK, Suzuki W, Schroeter H, Wall M, Sloan RP et al (2014) Enhancing dentate gyrus function with dietary flavanols improves cognition in older adults. Nat Neurosci 17:1798–1803CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Shors TJ, Miesegaes G, Beylin A, Zhao M, Rydel T, Gould E (2001) Neurogenesis in the adult is involved in the formation of trace memories. Nature 410:372–376CrossRefPubMedGoogle Scholar
  32. 32.
    Richetin K, Leclerc C, Toni N, Gallopin T, Pech S, Roybon L, Rampon C (2015) Genetic manipulation of adult-born hippocampal neurons rescues memory in a mouse model of Alzheimer’s disease. Brain 138:440–455CrossRefPubMedGoogle Scholar
  33. 33.
    Jinno S (2015) Aging affects new cell production in the adult hippocampus: A quantitative anatomic review. J Chem NeuroanatGoogle Scholar
  34. 34.
    Takeda A, Takada S, Nakamura M, Suzuki M, Tamano H, Ando M, Oku N (2011) Transient increase in Zn2+ in hippocampal CA1 pyramidal neurons causes reversible memory deficit. PLoS One 6:e28615CrossRefPubMedPubMedCentralGoogle Scholar
  35. 35.
    Liu S, Lau L, Wei J, Zhu D, Zou S, Sun HS, Fu Y, Liu F et al (2004) Expression of Ca(2+)-permeable AMPA receptor channels primes cell death in transient forebrain ischemia. Neuron 43:43–55CrossRefPubMedGoogle Scholar
  36. 36.
    Noh KM, Yokota H, Mashiko T, Castillo PE, Zukin RS, Bennett MV (2005) Blockade of calcium-permeable AMPA receptors protects hippocampal neurons against global ischemia-induced death. Proc Nat Acad Sci USA 102:12230–12235CrossRefPubMedPubMedCentralGoogle Scholar
  37. 37.
    Weiss JH (2011) Ca permeable AMPA channels in diseases of the nervous system. Front Mol Neurosci 4:42CrossRefPubMedPubMedCentralGoogle Scholar
  38. 38.
    Pagliusi SR, Gerrard P, Abdallah M, Talabot D, Catsicas S (1994) Age-related changes in expression of AMPA-selective glutamate receptor subunits: Is calcium-permeability altered in hippocampal neurons? Neuroscience 61:429–433CrossRefPubMedGoogle Scholar
  39. 39.
    Thibault O, Landfield PW (1996) Increase in single L-type calcium channels in hippocampal neurons during aging. Science 272:1017–1020CrossRefPubMedGoogle Scholar
  40. 40.
    Krueger JN, Moore SJ, Parent R, McKinney BC, Lee A, Murphy GG (2016) A novel mouse model of the aged brain: over-expression of the L-type voltage-gated calcium channel CaV1.3 Behav Brain ResGoogle Scholar
  41. 41.
    Frederickson CJ, Giblin LJ, Krezel A, McAdoo DJ, Muelle RN, Zeng Y, Balaji RV, Masalha R et al (2006) Concentrations of extracellular free zinc (pZn)e in the central nervous system during simple anesthetization, ischemia and reperfusion. Exp Neurol 198:285–293CrossRefPubMedGoogle Scholar
  42. 42.
    Tamano H, Nishio R, Shakushi Y, Sasaki M, Koike Y, Osawa M, Takeda A (2017) In vitro and in vivo physiology of low nanomolar concentrations of Zn2+ in artificial cerebrospinal fluid. Sci Rep 7:42897. doi: 10.1038/srep42897 CrossRefPubMedPubMedCentralGoogle Scholar
  43. 43.
    Sensi SL, Canzoniero LM, Yu SP, Ying HS, Koh JY, Kerchner GA, Choi DW (1997) Measurement of intracellular free zinc in living cortical neurons: routes of entry. J Neurosci 17:9554–9564CrossRefPubMedGoogle Scholar
  44. 44.
    Colvin RA, Bush AI, Volitakis I, Fontaine CP, Thomas D, Kikuchi K, Holmes WR (2008) Insights into Zn2+ homeostasis in neurons from experimental and modeling studies. Am J Physiol Cell Physiol 294:C726–C742CrossRefPubMedGoogle Scholar
  45. 45.
    Nafstad PH (1967) An electron microscope study on the termination of the perforant path fibres in the hippocampus and the fascia dentata. Z Zellforsch Mikrosk Anat 76:532–542CrossRefPubMedGoogle Scholar
  46. 46.
    Zipp F, Nitsch R, Soriano E, Frotscher M (1989) Entorhinal fibers form synaptic contacts on parvalbumin-immunoreactive neurons in the rat fascia dentata. Brain Res 495:161–166CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  1. 1.Department of Neurophysiology, School of Pharmaceutical SciencesUniversity of ShizuokaShizuokaJapan

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