Neurochemical Research

, Volume 33, Issue 6, pp 1138–1144

Effect of Melatonin and Melatonylvalpromide on β-amyloid and Neurofilaments in N2a Cells

  • Xiao-Chuan Wang
  • Yin-Chun Zhang
  • Nithiananda Chatterjie
  • Inge Grundke-Iqbal
  • Khalid Iqbal
  • Jian-Zhi Wang
Original Paper

Abstract

In the present study, we have studied the effect of melatonin (Mt) and melatonin derivative, i.e., melatonylvalpromide (Mtv), on cell viability, β-amyloid (Aβ) production, cell morphology, and expression and phosphorylation of neurofilament proteins in wild-type murine neuroblastoma N2a (N2a/wt) and N2a stably transfected with amyloid precursor protein (N2a/APP) cell lines. The study used MTT assay, Sandwich ELISA, immunocytochemistry and Western blots techniques. The results showed that both Mt and Mtv could increase cell viability, but Mtv did so more effectively. The N2a/APP showed shorter and less amount of cell processes than N2a/wt, and Mtv but not Mt slightly improved the morphological changes in N2A/APP. Both Mt and Mtv suppressed the Aβ level in cell lysates, but the effect of Mtv was stronger than Mt. The immunoreaction to the non-phosphorylated neurofilament proteins probed by SMI32 and SMI33 were remarkably weaker in N2a/APP than N2a/wt, while the immunoreaction to the phosphorylated neurofilament proteins at SMI34 epitopes was slightly stronger in N2a/APP than N2a/wt, suggesting higher phosphorylation level of neurofilament proteins in N2a/APP. Treatment of the cells with Mt and Mtv increased the immunoreaction at SMI32 and SMI33 epitopes, while only Mtv but not Mt decreased the staining at SMI34 epitope, suggesting both Mt and Mtv promote dephosphorylation of neurofilament at SMI32 and SMI33 epitopes, while Mtv stimulates dephosphorylation of neurofilament at SMI34 epitope. These results suggest that Mtv may be a better candidate in arresting the intracellular accumulation of Aβ and protecting the cells from Aβ-related toxicity.

Keywords

Alzheimer’s disease β-Amyloid Neurofilament Melatonin Melatonylvalproylamide 

References

  1. 1.
    Armstrong RA (2006) Plaques and tangles and the pathogenesis of Alzheimer’s disease. Folia Neuropathol 44:1–11PubMedGoogle Scholar
  2. 2.
    Selkoe DJ (2002) Alzheimer’s disease is a synaptic failure. Science 298:789–791PubMedCrossRefGoogle Scholar
  3. 3.
    Iwatsubo T, Odaka A, Suzuki N, Mizusawa H, Nukina N, Ihara Y (1994) Visualization of Aβ42(43) and Aβ40 in senile plaques with end-specific Aβ monoclonals: evidence that an initially deposited species is Aβ42(43). Neuron 13:45–53PubMedCrossRefGoogle Scholar
  4. 4.
    Roher A, Wolfe D, Palutke M, KuKuruga D (1998) Purification, ultrastructure, and chemical analysis of Alzheimer disease amyloid plaque core protein. Proc Natl Acad Sci USA 83:2662–2666CrossRefGoogle Scholar
  5. 5.
    Cai XD, Golde TE, Younkin SG (1993) Release of excess amyloid beta protein from a mutant amyloid beta protein precursor. Science 259:514–516PubMedCrossRefGoogle Scholar
  6. 6.
    Citron M, Oltersdorf T, Haass C, McConlogue L, Hung AY, Selkoe DJ (1992) Mutation of the beta-amyloid precursor protein in familial Alzheimer’s disease increases betaprotein production. Nature 360:672–674PubMedCrossRefGoogle Scholar
  7. 7.
    Citron M, Teplow DB, Selkoe DJ (1995) Generation of amyloid beta protein from its precursor is sequence specific. Neuron 14:661–670PubMedCrossRefGoogle Scholar
  8. 8.
    Hardy J (1999) The shorter amyloid cascade hypothesis. Neurobiol Aging 20:85PubMedCrossRefGoogle Scholar
  9. 9.
    Grundke-Iqbal I, Iqbal K, Quinlan M, Tung YC, Zaidi MS, Wisniewski HM (1986) Microtubule-associated protein tau, a component of paired helical filaments. J Biol Chem 261:6084–6089PubMedGoogle Scholar
  10. 10.
    Drubin DG, Kirschner MW (1986) Tau protein function in living cells. J Cell Biol 103:2739–2746PubMedCrossRefGoogle Scholar
  11. 11.
    Willard M, Simon C (1981) Antibody decoration of neurofilament. Cell Biol 89:198–205CrossRefGoogle Scholar
  12. 12.
    Wang JZ, Tung YC, Wang YP, Li XT, Iqbal K, Grundke-Iqbal I (2001) Hyperphosphorylation and accumulation of neurofilament proteins in Alzheimer disease brain and in okadaic acid-treated SY5Y cells. FEBS Lett 507:81–87PubMedCrossRefGoogle Scholar
  13. 13.
    Hu YY, He SS, Wang XC, Grundke-Iqbal I, Iqbal K, Wang JZ (2002) Elevated level of phosphorylated neurofilament in CSF of AD. Neurosci Lett 320:156–160PubMedCrossRefGoogle Scholar
  14. 14.
    Kittur S, Hoh J, Endo H, Tourtellotte W, Weeks BS, Markesbery W (1994) Cytoskeletal neurofilament gene expression in brain tissue from Adpatients. I. Decrease in NF-L and NF-M message. J Geriatr Psychiatry Neurol 7:153–158PubMedGoogle Scholar
  15. 15.
    Magri F, Sarra S, Cinchetti W, Guazzoni V, Fioravanti M, Cravello L (2004) Qualitative and quantitative changes of melatonin levels in physiological and pathological aging and in centenarians. J Pineal Res 36:256–261PubMedCrossRefGoogle Scholar
  16. 16.
    Lahiri DK, Ge YW, Sharman EH, Bondy SC (2004) Age-related changes in serum melatonin in mice: higher levels of combined melatonin and 6-hydroxymelatonin sulfate in the cerebral cortex than serum, heart, liver and kidney tissues. J Pineal Res 36:217–223PubMedCrossRefGoogle Scholar
  17. 17.
    Liu RY, Zhou JN, van Heerikhuize J, Hofman MA, Swaab DF (1999) Decreased melatonin levels in postmortem cerebrospinal fluid in relation to aging, Alzheimer’s disease, and apolipoprotein E-epsilon4/4 genotype. J Clin Endocrinol Metab 84:323–327PubMedCrossRefGoogle Scholar
  18. 18.
    Wu YH, Feenstra MG, Zhou JN, Liu RY, Torano JS, Van Kan HJ (2003) Molecular changes underlying reduced pineal melatonin levels in Alzheimer disease: alterations in preclinical and clinical stages. J Clin Endocrinol Metab 88:5898–5906PubMedCrossRefGoogle Scholar
  19. 19.
    Pappolla M, Bozner P, Soto C, Shao HY, Robakis NK, Zagorski M (1998) Inhibition of Alzheimer β-fibrillogengsis by melatonin. J Biol Chem 273:7185–7188PubMedCrossRefGoogle Scholar
  20. 20.
    Pappolla MA, Chyan YJ, Poeggeler B, Frangione B, Wilson G, Ghiso J (2000) An assessment of the antioxidant and the antiamyloidogenic properties of melatonin: implications for Alzheimer’s disease. J Neural Transm 107:203–231PubMedCrossRefGoogle Scholar
  21. 21.
    Castro LM, Gallant M, Niles LP (2005) Novel targets for valproic acid: up-regulation of melatonin receptors and neurotrophic factors in C6 glioma cells. J Neurochem 95:1227–1236PubMedCrossRefGoogle Scholar
  22. 22.
    Chatterjie N, Alexander G, Wang H (2001) Synthesis of valproic acid amides of a melatonin derivative, a piracetam and amantadine for biological tests. Neurochem Res 26:1171–1176PubMedCrossRefGoogle Scholar
  23. 23.
    Yamamoto T, Yuyama K, Nakamura K, Kato T, Yamamoto H (2000) Kinetic characterization of the nitric oxide toxicity for pc12 cells: effect of half-life time of no release. Eur J Pharmacol 397:25–33PubMedCrossRefGoogle Scholar
  24. 24.
    Turner RS, Suzuki N, Chyung AS, Younkin SG, Lee VM (1996) Amyloids β40 and β42 are generated intracellularly in cultured human neurons and their secretion increases with maturation. J Biol Chem 271:8966–8970PubMedCrossRefGoogle Scholar
  25. 25.
    Zhang YC, Wang ZF, Wang Q, Wang YP, Wang JZ (2004) Melatonin attenuates overproduction of β-amyloid-induced inhibition in expression of neurofilament protein. Acta Pharmacol Sin 25:447–451PubMedGoogle Scholar
  26. 26.
    Sternberger NH, Sternberger LA, Ulrich J (1985) Aberrant neurofilament phosphorylation in Alzheimer disease. PNAS 82:4274–4276PubMedCrossRefGoogle Scholar
  27. 27.
    Wang DL, Ling ZQ, Cao FY, Zhu LQ, Wang JZ (2004) Melatonin attenuates isoproterenol-induced protein kinase A overactivation and tau hyperphosphorylation in rat brain. J Pineal Res 37:11–16PubMedCrossRefGoogle Scholar
  28. 28.
    Selkoe DJ (1996) Amyloid β-protein and the genetics of Alzheimer’s disease. J Biol Chem 271:18295–18298PubMedGoogle Scholar
  29. 29.
    Gouras GK, Almeida CG, Takahashi RH (2005) Intraneuronal Aβ accumulation and origin of plaques in Alzheimer’s disease. Neurobiol Aging 26:1235–1244PubMedCrossRefGoogle Scholar
  30. 30.
    Tickler AK, Wade JD, Separovic F (2005) The role of Aβ peptides in Alzheimer’s disease. Protein Pept Lett 12:513–519PubMedCrossRefGoogle Scholar
  31. 31.
    Yang Y, Dowling J, Yu QC, Kouklis P, Cleveland DW, Fuchs E (1996) An essential cytoskeleton linker protein connecting actin microfilaments to intermediate filaments. Cell 86:655–665PubMedCrossRefGoogle Scholar
  32. 32.
    Ferrari E, Arcaini A, Gornati R, Pelanconi L, Cravello L, Fioravanti M (2000) Pineal and pituitary-adrenocortical function in physiological aging and in senile dementia. Exp Gerontol 35:1239–1250PubMedCrossRefGoogle Scholar
  33. 33.
    Maurizi CP (2001) Alzheimer’s disease: roles for mitochondrial damage, the hydroxyl radical, and cerebrospinal fluid deficiency of melatonin. Med Hypotheses 57:156–160PubMedCrossRefGoogle Scholar
  34. 34.
    Li SP, Deng YQ, Wang XC, Wang YP, Wang JZ (2004) Melatonin protects SH-SY5Y neuroblastoma cells from calyculin A-induced neurofilament impairment and neurotoxicity. J Pineal Res 36:186–191PubMedCrossRefGoogle Scholar
  35. 35.
    Li XC, Wang ZF, Zhang JX, Wang Q, Wang JZ (2005) Effect of melatonin on calyculin A-induced tau hyperphosphorylation. Eur J Pharmacol 510:25–30PubMedCrossRefGoogle Scholar
  36. 36.
    Deng YQ, Xu GG, Duan P, Zhang Q, Wang JZ (2005) Effects of melatonin on wortmannin-induced tau hyperphosphorylation. Acta Pharmacol Sin 26:519–526PubMedCrossRefGoogle Scholar
  37. 37.
    Wang JZ, Wang ZF (2006) Role of melatonin in Alzheimer-like neurodegeneration. Acta Pharmacol Sin 27:41–49PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2007

Authors and Affiliations

  • Xiao-Chuan Wang
    • 1
  • Yin-Chun Zhang
    • 1
  • Nithiananda Chatterjie
    • 2
  • Inge Grundke-Iqbal
    • 2
  • Khalid Iqbal
    • 2
  • Jian-Zhi Wang
    • 1
  1. 1.Department of Pathophysiology, Hubei Provincial Key Laboratory in Neurological Diseases, Tongji Medical CollegeHuazhong University of Science and TechnologyWuhanP.R.China
  2. 2.Department of NeurochemistryNYS Institute for Basic ResearchStaten IslandUSA

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