Cellular and Molecular Neurobiology

, Volume 38, Issue 4, pp 817–826 | Cite as

Effects of Acetylcholine on β-Amyloid-Induced cPLA2 Activation in the TB Neuroectodermal Cell Line: Implications for the Pathogenesis of Alzheimer’s Disease

  • Arianna Polverino
  • Manuela Grimaldi
  • Pierpaolo Sorrentino
  • Francesca Jacini
  • Anna Maria D’Ursi
  • Giuseppe Sorrentino
Original Research
  • 100 Downloads

Abstract

The role of β-amyloid (Aβ) in the pathogenesis of Alzheimer’s disease (AD) is still considered crucial. The state of Aβ aggregation is critical in promoting neuronal loss and neuronal function impairment. Recently, we demonstrated that Acetylcholine (ACh) is neuroprotective against the toxic effects of Aβ in the cholinergic LAN-2 cells. In biophysical experiments, ACh promotes the soluble Aβ peptide conformation rather than the aggregation-prone β-sheet conformation. In order to better understand the biological role of ACh in AD, we studied the effect of Aβ on the phosphorylation of the cytosolic phospholipase A2 (cPLA2) in the TB neuroectodermal cell line, which differentiates toward a neuronal phenotype when cultured in the presence of retinoic acid (RA). We chose the phosphorylated form of cPLA2 (Ser505, Phospho-cPLA2) as a biomarker to test the influence of ACh on the effects of Aβ in both undifferentiated and RA-differentiated TB cells. Our results show that TB cells are responsive to Aβ. Moreover, in undifferentiated cells 1 h treatment with Aβ induces a 2.5-fold increase of the Phospho-cPLA2 level compared to the control after 24 h in vitro, while no significant difference is observed between Aβ-treated and non-treated cells after 4 and 7 days in vitro. The RA-differentiated cells are not sensitive to Aβ. In TB cell line ACh is able to blunt the effects of Aβ. The ability of ACh to protect non-cholinergic cells against Aβ reinforces the hypothesis that, in addition to its role in cholinergic transmission, ACh could also act as a neuroprotective agent.

Keywords

Alzheimer’s disease β-Amyloid Phospholipase A2 Acetylcholine TB cell line Differentiation 

Notes

Acknowledgements

The authors thank Dr. Maria Teodora Valente and Dr. Luisa Arnese for their helpful technical support during the experiments.

Funding

The work was supported by a Grant from MIUR (FIRB–MERIT RBNE08LN4P:006) and University of Naples Parthenope, Naples, Italy.

Author Contributions

Authors made substantial contributions to conception and design, acquisition of data, analysis, and interpretation of data. Authors participated in drafting the article or revising it critically and they gave final approval of the version to be submitted. Study conception and design A. Polverino, A.M. D’Ursi, G. Sorrentino; Acquisition of data A. Polverino, M. Grimaldi; Analysis and interpretation of data A. Polverino, M. Grimaldi, P. Sorrentino, F. Jacini, A.M. D’Ursi, G. Sorrentino; Drafting of manuscript A. Polverino, M. Grimaldi, P. Sorrentino, F. Jacini, A.M. D’Ursi, G. Sorrentino; English Quality revision P. Sorrentino; Critical revision G. Sorrentino, A. Polverino.

Compliance with Ethical Standards

Conflict of interest

The authors declare that there is no personal or institutional conflict of interest related to the presented research and its publication.

Ethical Approval

All procedures performed in studies involving human were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments.

Informed Consent

Informed consent was obtained from the relatives of the patient involved in the study.

Supplementary material

10571_2017_555_MOESM1_ESM.pdf (3.2 mb)
Supplementary material 1 (PDF 3302 kb)

References

  1. Abramov AY, Canevari L, Duchen MR (2004) Calcium signals induced by amyloid β peptide and their consequences in neurons and astrocytes in culture. Biochimica et Biophys Acta 1742:81–87CrossRefGoogle Scholar
  2. Baglioni S et al (2006) Prefibrillar amyloid aggregates could be generic toxins in higher organisms. J Neurosci 26:8160–8167CrossRefPubMedGoogle Scholar
  3. Bartus RT (2000) On neurodegenerative diseases, models, and treatment strategies: lessons learned and lessons forgotten a generation following the cholinergic hypothesis. Exp Neurol 163:495–529CrossRefPubMedGoogle Scholar
  4. Bate C, Williams A (2010) Amyloid-β1–40 inhibits amyloid-β1–42 induced activation of cytoplasmic phospholipase A2 and synapse degeneration. J Alzheimer’s Dis 21:985–993CrossRefGoogle Scholar
  5. Campanari M-L, Navarrete F, Ginsberg SD, Manzanares J, Sáez-Valero J, García-Ayllón M-S (2016) Increased expression of readthrough acetylcholinesterase variants in the brains of Alzheimer’s disease patients. J Alzheimer’s Dis 53:831–841CrossRefGoogle Scholar
  6. Cecchi C et al (2008) Replicating neuroblastoma cells in different cell cycle phases display different vulnerability to amyloid toxicity. J Mol Med 86:197–209CrossRefPubMedGoogle Scholar
  7. Chan WC, White PD (2000) Fmoc solid phase peptide synthesis. Oxford University Press, OxfordGoogle Scholar
  8. Chan MC, Bautista E, Alvarado-Cruz I, Quintanilla-Vega B, Segovia J (2017) Inorganic mercury prevents the differentiation of SH-SY5Y cells: amyloid precursor protein, microtubule associated proteins and ROS as potential targets. J Trace Elem Med Biol 41:119–128CrossRefPubMedGoogle Scholar
  9. Craig LA, Hong NS, McDonald RJ (2011) Revisiting the cholinergic hypothesis in the development of Alzheimer’s disease. Neurosci Biobehav Rev 35:1397–1409CrossRefPubMedGoogle Scholar
  10. Davanzo C, Aronson J, Kim YH, Choi SH, Tanzi RE, Kim DY (2015) Alzheimer’s in 3D culture: challenges and perspectives. Bioessays 37:1139–1148CrossRefGoogle Scholar
  11. Davies P, Maloney A (1976) Selective loss of central cholinergic neurons in Alzheimer’s disease. Lancet 308:1403CrossRefGoogle Scholar
  12. Desbène C et al (2012) Critical role of cPLA 2 in Aβ oligomer-induced neurodegeneration and memory deficit. Neurobiol Aging 33:1123CrossRefPubMedGoogle Scholar
  13. Fiandaca MS, Mapstone ME, Cheema AK, Federoff HJ (2014) The critical need for defining preclinical biomarkers in Alzheimer’s disease. Alzheimer’s Dement 10:S196–S212CrossRefGoogle Scholar
  14. Fonseca MB, Solá S, Xavier JM, Dionísio PA, Rodrigues CM (2013) Amyloid β peptides promote autophagy-dependent differentiation of mouse neural stem cells. Mol Neurobiol 48:829–840CrossRefPubMedGoogle Scholar
  15. Grimaldi M et al (2016) β-Amyloid-acetylcholine molecular interaction: new role of cholinergic mediators in anti-Alzheimer therapy? Future Med Chem 8:1179–1189CrossRefPubMedGoogle Scholar
  16. Haass C, Selkoe DJ (2007) Soluble protein oligomers in neurodegeneration: lessons from the Alzheimer’s amyloid β-peptide Nature reviews. Mol Cell Biol 8:101–112Google Scholar
  17. Hardy JA, Higgins GA (1992) Alzheimer’s disease: the amyloid cascade hypothesis. Science 256:184CrossRefPubMedGoogle Scholar
  18. Hughes E, Burke RM, Doig AJ (2000) Inhibition of Toxicity in the β-Amyloid Peptide Fragment β-(25–35) Using N-Methylated Derivatives A GENERAL STRATEGY TO PREVENT AMYLOID FORMATION. J Biol Chem 275:25109–25115CrossRefPubMedGoogle Scholar
  19. Kaminsky YG, Marlatt MW, Smith MA, Kosenko EA (2010) Subcellular and metabolic examination of amyloid-β peptides in Alzheimer disease pathogenesis: evidence for Aβ 25–35. Exp Neurol 221:26–37CrossRefPubMedGoogle Scholar
  20. Kanfer JN, Sorrentino G, Sitar DS (1998) Phospholipases as mediators of amyloid beta peptide neurotoxicity: an early event contributing to neurodegeneration characteristic of Alzheimer’s disease. Neurosci Lett 257:93–96CrossRefPubMedGoogle Scholar
  21. Kanfer JN, Sorrentino G, Sitar DS (1999) Amyloid beta peptide membrane perturbation is the basis for its biological effects. Neurochem Res 24:1621–1630CrossRefPubMedGoogle Scholar
  22. Kang J et al (1987) The precursor of Alzheimer’s disease amyloid A4 protein resembles a cell-surface receptor. Nature 325:733–736CrossRefPubMedGoogle Scholar
  23. Kar S, Seto D, Gaudreau P, Quirion R (1996) Beta-amyloid-related peptides inhibit potassium-evoked acetylcholine release from rat hippocampal slices. J Neurosci 16:1034–1040CrossRefPubMedGoogle Scholar
  24. Kayed R et al (2009) Annular protofibrils are a structurally and functionally distinct type of amyloid oligomer. J Biol Chem 284:4230–4237CrossRefPubMedPubMedCentralGoogle Scholar
  25. Kumar R, Nordberg A, Darreh-Shori T (2015) Amyloid-β peptides act as allosteric modulators of cholinergic signalling through formation of soluble BAβACs. Brain 139:174–192CrossRefPubMedPubMedCentralGoogle Scholar
  26. Kwak Y-D et al (2006) Amyloid precursor protein regulates differentiation of human neural stem cells. Stem Cells Dev 15:381–389CrossRefPubMedGoogle Scholar
  27. Lee JCM, Simonyi A, Sun AY, Sun GY (2011) Phospholipases A2 and neural membrane dynamics: implications for Alzheimer’s disease. J Neurochem 116:813–819CrossRefPubMedPubMedCentralGoogle Scholar
  28. Mapstone M et al (2014) Plasma phospholipids identify antecedent memory impairment in older adults. Nat Med 20:415–418CrossRefPubMedPubMedCentralGoogle Scholar
  29. Marutle A, Ohmitsu M, Nilbratt M, Greig NH, Nordberg A, Sugaya K (2007) Modulation of human neural stem cell differentiation in Alzheimer (APP23) transgenic mice by phenserine. Proc Natl Acad Sci USA 104:12506–12511CrossRefPubMedPubMedCentralGoogle Scholar
  30. Naldi M et al (2012) Amyloid β-peptide 25–35 self-assembly and its inhibition: a model undecapeptide system to gain atomistic and secondary structure details of the Alzheimer’s disease process and treatment. ACS Chem Neurosci 3:952–962CrossRefPubMedPubMedCentralGoogle Scholar
  31. Pimplikar SW, Nixon RA, Robakis NK, Shen J, Tsai L-H (2010) Amyloid-independent mechanisms in Alzheimer’s disease pathogenesis. J Neurosci 30:14946–14954CrossRefPubMedPubMedCentralGoogle Scholar
  32. Portelius E et al (2006) An Alzheimer’s disease-specific β-amyloid fragment signature in cerebrospinal fluid. Neurosci Lett 409:215–219CrossRefPubMedGoogle Scholar
  33. Rancic A, Filipovic N, Lovric JM, Mardesic S, Saraga-Babic M, Vukojevic K (2017) Neuronal differentiation in the early human retinogenesis. Acta Histochem 119:264–272CrossRefPubMedGoogle Scholar
  34. Rönicke R, Klemm A, Meinhardt J, Schröder UH, Fändrich M, Reymann KG (2008) Aβ mediated diminution of MTT reduction—an artefact of single cell culture? PLoS ONE 3:e3236CrossRefPubMedPubMedCentralGoogle Scholar
  35. Sanchez-Mejia RO et al (2008) Phospholipase A2 reduction ameliorates cognitive deficits in a mouse model of Alzheimer’s disease. Nat Neurosci 11:1311–1318CrossRefPubMedPubMedCentralGoogle Scholar
  36. Selkoe DJ (2001) Alzheimer’s disease: genes, proteins, and therapy. Physiol Rev 81:741–766CrossRefPubMedGoogle Scholar
  37. Singh I, Sorrentino G, McCartney D, Massarelli R, Kanfer J (1990) Enzymatic activites during differentiation of the human neuroblastoma cells, LA-N-1 and LA-N-2. J Neurosci Res 25:476–485CrossRefPubMedGoogle Scholar
  38. Singh I, Sorrentino G, Sitar D, Kanfer J (1997) Indomethacin and nordihydroguaiaretic acid inhibition of amyloid β protein (25–35) activation of phospholipases A 2 and D of LA-N-2 cells. Neurosci Lett 222:5–8CrossRefPubMedGoogle Scholar
  39. Sorrentino G, Monsurrò MR, Pettinato G, Vanni R, Zuddas A, Di Porzio U, Bonavita V (1999) Establishment and characterization of a human neuroectodermal cell line (TB) from a cerebrospinal fluid specimen. Brain Res 827:205–209CrossRefPubMedGoogle Scholar
  40. Sorrentino G, Migliaccio R, Bonavita V (2008) Treatment of vascular dementia: the route of prevention. Eur Neurol 60:217CrossRefPubMedGoogle Scholar
  41. Sorrentino P, Iuliano A, Polverino A, Jacini F, Sorrentino G (2014) The dark sides of amyloid in Alzheimer’s disease pathogenesis. FEBS Lett 588:641–652CrossRefPubMedGoogle Scholar
  42. Stains CI, Mondal K, Ghosh I (2007) Molecules that Target beta-Amyloid. ChemMedChem 2:1674–1692CrossRefPubMedGoogle Scholar
  43. Suebsoonthron J, Jaroonwitchawan T, Yamabhai M, Noisa P (2017) Inhibition of WNT signaling reduces differentiation and induces sensitivity to doxorubicin in human malignant neuroblastoma SH-SY5Y cells. Anticancer Drugs 28:469–479CrossRefPubMedGoogle Scholar
  44. Sun GY, Xu J, Jensen MD, Simonyi A (2004) Phospholipase A2 in the central nervous system implications for neurodegenerative diseases. J Lipid Res 45:205–213CrossRefPubMedGoogle Scholar
  45. Vaucher E, Aumont N, Pearson D, Rowe W, Poirier J, Kar S (2001) Amyloid β peptide levels and its effects on hippocampal acetylcholine release in aged, cognitively-impaired and-unimpaired rats. J Chem Neuroanat 21:323–329CrossRefPubMedGoogle Scholar
  46. Wattmo C, Wallin ÅK, Minthon L (2012) Functional response to cholinesterase inhibitor therapy in a naturalistic Alzheimer’s disease cohort. BMC Neurol 12:134CrossRefPubMedPubMedCentralGoogle Scholar
  47. Weidemann A, König G, Bunke D, Fischer P, Salbaum JM, Masters CL, Beyreuther K (1989) Identification, biogenesis, and localization of precursors of Alzheimer’s disease A4 amyloid protein. Cell 57:115–126CrossRefPubMedGoogle Scholar
  48. Zhu D, Lai Y, Shelat PB, Hu C, Sun GY, Lee JC (2006) Phospholipases A2 mediate amyloid-β peptide-induced mitochondrial dysfunction. J Neurosci 26:11111–11119CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2017

Authors and Affiliations

  • Arianna Polverino
    • 1
    • 2
  • Manuela Grimaldi
    • 3
  • Pierpaolo Sorrentino
    • 4
  • Francesca Jacini
    • 1
  • Anna Maria D’Ursi
    • 3
  • Giuseppe Sorrentino
    • 1
    • 2
  1. 1.Department of Motor Sciences and WellnessUniversity of Naples ParthenopeNaplesItaly
  2. 2.Institute of Diagnosis and Treatment HermitageNaplesItaly
  3. 3.Department of PharmacyUniversity of SalernoFisciano, SalernoItaly
  4. 4.Department of EngineeringUniversity of Naples ParthenopeNaplesItaly

Personalised recommendations