Expression levels of the α7 nicotinic acetylcholine receptor in the brains of patients with Alzheimer's disease and their effect on synaptic proteins in SH-SY5Y cells

Ren,Jia-Mou; Zhang,Shu-Li; Wang,Xiao-Ling; Guan,Zhi-Zhong; Qi,Xiao-Lan

doi:10.3892/mmr.2020.11253

Expression levels of the α7 nicotinic acetylcholine receptor in the brains of patients with Alzheimer's disease and their effect on synaptic proteins in SH-SY5Y cells

Authors:
- Jia-Mou Ren
- Shu-Li Zhang
- Xiao-Ling Wang
- Zhi-Zhong Guan
- Xiao-Lan Qi
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Affiliations: Key Laboratory of Endemic and Ethnic Diseases, Ministry of Education, Guizhou Medical University, Guiyang, Guizhou 550004, P.R. China
Published online on: June 18, 2020 https://doi.org/10.3892/mmr.2020.11253
Pages: 2063-2075
Copyright: © Ren et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Alzheimer's disease (AD) is a chronic neurodegenerative, and abnormal aggregation of the neurotoxic β amyloid (Aβ) peptide is an early event in AD. The present study aimed to determine the correlation between the nicotinic acetylcholine receptor α7 subunit (α7 nAChR) and Aβ in the brains of patients with AD, and to investigate whether the increased expression levels of the α7 nAChR could alter the neurotoxicity of Aβ. The expression levels of α7 nAChR and Aβ in the brains of patients with AD and healthy brains were analyzed using immunofluorescence. Moreover, SH‑SY5Y cells were used to stably overexpress or silence α7 nAChR expression levels, prior to the treatment with or without 1 µmol/l Aβ1‑42 oligomer (AβO). The mRNA and protein expression levels of α7 nAChR, synaptophysin (SYP), postsynaptic density of 95 kDa (PSD‑95) and synaptosomal‑associated protein of 25 kDa (SNAP‑25) were subsequently analyzed using reverse transcription‑quantitative PCR and western blotting. In addition, the concentration of acetylcholine (ACh) and the activity of acetylcholinesterase (AChE) were analyzed using spectrophotometry, while the cell apoptotic rate was determined using flow cytometry. The expression of Aβ in the brains of patients with AD was found to be significantly increased, whereas the expression of α7 nAChR was significantly decreased compared with the healthy control group. In vitro, the expression levels of α7 nAChR were significantly increased or decreased following the overexpression or silencing of the gene, respectively. Consistent with these observations, the mRNA and protein expression levels of SYP, PSD‑95 and SNAP‑25 were also significantly increased following the overexpression of α7 nAChR and decreased following the genetic silencing of the receptor. In untransfected or negative control cells, the expression levels of these factors and the apoptotic rate were significantly reduced following the exposure to AβO, which was found to be attenuated by α7 nAChR overexpression, but potentiated by α7 nAChR RNA silencing. However, no significant differences were observed in either the ACh concentration or AChE activity following transfection. Collectively, these findings suggested that α7 nAChR may protect the brains of patients with AD against Aβ, as α7 nAChR overexpression increased the expression levels of SYP, SNAP‑25 and PSD‑95, and attenuated the inhibitory effect of Aβ on the expression of these synaptic proteins and cell apoptosis. Overall, this indicated that α7 nAChR may serve an important neuroprotective role in AD.

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1	Bloom GS: Amyloid-β and tau: The trigger and bullet in Alzheimer disease pathogenesis. JAMA Neurol. 71:505–508. 2014. View Article : Google Scholar : PubMed/NCBI
2	Amemori T, Jendelova P, Ruzicka J, Urdzikova LM and Sykova E: Alzheimer's disease: Mechanism and approach to cell therapy. Int J Mol Sci. 16:26417–26451. 2015. View Article : Google Scholar : PubMed/NCBI
3	Ferreira ST, Lourenco MV, Oliveira MM and De Felice FG: Soluble amyloid-β oligomers as synaptotoxins leading to cognitive impairment in Alzheimer's disease. Front Cell Neurosci. 9:1912015. View Article : Google Scholar : PubMed/NCBI
4	Viola KL and Klein WL: Amyloid β oligomers in Alzheimer's disease pathogenesis, treatment, and diagnosis. Acta Neuropathol. 129:183–206. 2015. View Article : Google Scholar : PubMed/NCBI
5	Lesné SE, Sherman MA, Grant M, Kuskowski M, Schneider JA, Bennett DA and Ashe KH: Brain amyloid-β oligomers in ageing and Alzheimer's disease. Brain. 136:1383–1398. 2013. View Article : Google Scholar : PubMed/NCBI
6	Magi S, Castaldo P, Macrì ML, Maiolino M, Matteucci A, Bastioli G, Gratteri S, Amoroso S and Lariccia V: Intracellular calcium dysregulation: Implications for Alzheimer's disease. BioMed Res Int. 2016:67013242016. View Article : Google Scholar : PubMed/NCBI
7	Jagust W: Is amyloid-β harmful to the brain? Insights from human imaging studies. Brain. 139:23–30. 2016. View Article : Google Scholar : PubMed/NCBI
8	Overk CR and Masliah E: Pathogenesis of synaptic degeneration in Alzheimer's disease and Lewy body disease. Biochem Pharmacol. 88:508–516. 2014. View Article : Google Scholar : PubMed/NCBI
9	Sheng M, Sabatini BL and Südhof TC: Synapses and Alzheimer's disease. Csh Perspect Biol. 4:doi.org/10.1101/cshperspect.a005777.
10	Harrill JA, Chen H, Streifel KM, Yang D, Mundy WR and Lein PJ: Ontogeny of biochemical, morphological and functional parameters of synaptogenesis in primary cultures of rat hippocampal and cortical neurons. Mol Brain. 8:102015. View Article : Google Scholar : PubMed/NCBI
11	García-Morales V, Montero F, González-Forero D, Rodríguez-Bey G, Gómez-Pérez L, Medialdea-Wandossell MJ, Domínguez-Vías G, García-Verdugo JM and Moreno-López B: Membrane-derived phospholipids control synaptic neurotransmission and plasticity. PLoS Biol. 13:e10021532015. View Article : Google Scholar : PubMed/NCBI
12	Wang DB, Kinoshita Y, Kinoshita C, Uo T, Sopher BL, Cudaback E, Keene CD, Bilousova T, Gylys K, Case A, et al: Loss of endophilin-B1 exacerbates Alzheimer's disease pathology. Brain. 138:2005–2019. 2015. View Article : Google Scholar : PubMed/NCBI
13	Marcello E, Epis R, Saraceno C and Di Luca M: Synaptic dysfunction in Alzheimer's disease. Adv Exp Med Biol. 970:573–601. 2012. View Article : Google Scholar : PubMed/NCBI
14	Wang DB, Kinoshita Y, Kinoshita C, Uo T, Sopher BL, Cudaback E, Keene CD, Bilousova T, Gylys K, Case A, et al: Loss of endophilin-B1 exacerbates Alzheimer's disease pathology. Brain. 138:2005–2019. 2015. View Article : Google Scholar : PubMed/NCBI
15	Sivanesan S, Tan A and Rajadas J: Pathogenesis of Abeta oligomers in synaptic failure. Curr Alzheimer Res. 10:316–323. 2013. View Article : Google Scholar : PubMed/NCBI
16	Antonucci F, Corradini I, Fossati G, Tomasoni R, Menna E and Matteoli M: SNAP-25, a known presynaptic protein with emerging postsynaptic functions. Front Synaptic Neurosci. 8:72016. View Article : Google Scholar : PubMed/NCBI
17	Wang J, Yuan J, Pang J, Ma J, Han B, Geng Y, Shen L, Wang H, Ma Q, Wang Y and Wang M: Effects of chronic stress on cognition in male SAMP8 mice. Cell Physiol Biochem. 39:1078–1086. 2016. View Article : Google Scholar : PubMed/NCBI
18	Xi YD, Zhang DD, Ding J, Yu HL, Yuan LH, Ma WW, Han J and Xiao R: Genistein inhibits Aβ25-35-induced synaptic toxicity and regulates CaMKII/CREB pathway in SH-SY5Y cells. Cell Mol Neurobiol. 36:1151–1159. 2016. View Article : Google Scholar : PubMed/NCBI
19	Ito S, Ménard M, Atkinson T, Brown L, Whitfield J and Chakravarthy B: Relative expression of the p75 neurotrophin receptor, tyrosine receptor kinase A, and insulin receptor in SH-SY5Y neuroblastoma cells and hippocampi from Alzheimer's disease patients. Neurochem Int. 101:22–29. 2016. View Article : Google Scholar : PubMed/NCBI
20	Gray NE, Zweig JA, Kawamoto C, Quinn JF and Copenhaver PF: STX, a novel membrane estrogen receptor ligand, protects against amyloid-β toxicity. J Alzheimers Dis. 51:391–403. 2016. View Article : Google Scholar : PubMed/NCBI
21	Ferreira-Vieira TH, Guimaraes IM, Silva FR and Ribeiro FM: Alzheimer's disease: Targeting the cholinergic system. Curr Neuropharmacol. 14:101–115. 2016. View Article : Google Scholar : PubMed/NCBI
22	Park D, Choi EK, Cho TH, Joo SS and Kim YB: Human neural stem cells encoding ChAT gene restore cognitive function via acetylcholine synthesis, Aβ elimination, and neuroregeneration in APPswe/PS1dE9 mice. Int J Mol Sci. 21:212020. View Article : Google Scholar
23	Badin AS, Eraifej J and Greenfield S: High-resolution spatio-temporal bioactivity of a novel peptide revealed by optical imaging in rat orbitofrontal cortex in vitro: Possible implications for neurodegenerative diseases. Neuropharmacology. 73:10–18. 2013. View Article : Google Scholar : PubMed/NCBI
24	Galimberti D and Scarpini E: Old and new acetylcholinesterase inhibitors for Alzheimer's disease. Expert Opin Investig Drugs. 25:1181–1187. 2016. View Article : Google Scholar : PubMed/NCBI
25	Fukunaga K and Yabuki Y: SAK3-induced neuroprotection is mediated by nicotinic acetylcholine receptors. In: Nicotinic Acetylcholine Receptor Signaling in Neuroprotection. Akaike A, Shimohama S and Yoshimi Misu Y: Springer; Berlin: pp. 159–171. 2018, PubMed/NCBI
26	Hernandez CM, Kayed R, Zheng H, Sweatt JD and Dineley KT: Loss of alpha7 nicotinic receptors enhances beta-amyloid oligomer accumulation, exacerbating early-stage cognitive decline and septohippocampal pathology in a mouse model of Alzheimer's disease. J Neurosci. 30:2442–2453. 2010. View Article : Google Scholar : PubMed/NCBI
27	Gil SM and Metherate R: Enhanced sensory-cognitive processing by activation of nicotinic acetylcholine receptors. Nicotine Tob Res. 21:377–382. 2019. View Article : Google Scholar : PubMed/NCBI
28	Gu Z and Yakel JL: Timing-dependent septal cholinergic induction of dynamic hippocampal synaptic plasticity. Neuron. 71:155–165. 2011. View Article : Google Scholar : PubMed/NCBI
29	Lozada AF, Wang X, Gounko NV, Massey KA, Duan J, Liu Z and Berg DK: Glutamatergic synapse formation is promoted by α7-containing nicotinic acetylcholine receptors. J Neurosci. 32:7651–7661. 2012. View Article : Google Scholar : PubMed/NCBI
30	Yue Y, Liu R, Cheng W, Hu Y, Li J, Pan X, Peng J and Zhang P: GTS-21 attenuates lipopolysaccharide-induced inflammatory cytokine production in vitro by modulating the Akt and NF-κB signaling pathway through the α7 nicotinic acetylcholine receptor. Int Immunopharmacol. 29:504–512. 2015. View Article : Google Scholar : PubMed/NCBI
31	Tyagi E, Agrawal R, Nath C and Shukla R: Inhibitory role of cholinergic system mediated via alpha7 nicotinic acetylcholine receptor in LPS-induced neuro-inflammation. Innate Immun. 16:3–13. 2010. View Article : Google Scholar : PubMed/NCBI
32	Liao Y, Qi XL, Cao Y, Yu WF, Ravid R, Winblad B, Pei JJ and Guan ZZ: Elevations in the levels of NF-κB and inflammatory chemotactic factors in the brains with Alzheimer's disease - One mechanism may involve α3 nicotinic acetylcholine receptor. Curr Alzheimer Res. 13:1290–1301. 2016. View Article : Google Scholar : PubMed/NCBI
33	Domínguez-Álvaro M, Montero-Crespo M, Blazquez-Llorca L, Insausti R, DeFelipe J and Alonso-Nanclares L: Three-dimensional analysis of synapses in the transentorhinal cortex of Alzheimer's disease patients. Acta Neuropathol Commun. 6:202018. View Article : Google Scholar : PubMed/NCBI
34	Bauwens M, Mottaghy FM and Bucerius J: PET imaging of the human nicotinic cholinergic pathway in atherosclerosis. Curr Cardiol Rep. 17:672015. View Article : Google Scholar : PubMed/NCBI
35	Dubois B, Feldman HH, Jacova C, Dekosky ST, Barberger-Gateau P, Cummings J, Delacourte A, Galasko D, Gauthier S, Jicha G, et al: Research criteria for the diagnosis of Alzheimer's disease: Revising the NINCDS-ADRDA criteria. Lancet Neurol. 6:734–746. 2007. View Article : Google Scholar : PubMed/NCBI
36	McKhann G, Drachman D, Folstein M, Katzman R, Price D and Stadlan EM: Clinical diagnosis of Alzheimer's disease: Report of the NINCDS-ADRDA work group under the auspices of Department of Health and Human Services Task Force on Alzheimer's disease. Neurology. 34:939–944. 1984. View Article : Google Scholar : PubMed/NCBI
37	Reisberg B, Ferris SH, de Leon MJ and Crook T: The Global Deterioration Scale for assessment of primary degenerative dementia. Am J Psychiatry. 139:1136–1139. 1982. View Article : Google Scholar : PubMed/NCBI
38	Wang XL, Deng YX, Gao YM, Dong YT, Wang F, Guan ZZ, Wei H and Qi XL: Activation of α7 nAChR by PNU-282987 improves synaptic and cognitive functions through restoring the expression of synaptic-associated proteins and the CaM-CaMKII-CREB signaling pathway. Aging (Albany NY). 12:543–570. 2020. View Article : Google Scholar : PubMed/NCBI
39	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
40	Dunant Y and Gisiger V: Ultrafast and slow cholinergic transmission. Different involvement of acetylcholinesterase molecular forms. Molecules. 22:13002017. View Article : Google Scholar
41	Dani JA and Bertrand D: Nicotinic acetylcholine receptors and nicotinic cholinergic mechanisms of the central nervous system. Annu Rev Pharmacol Toxicol. 47:699–729. 2007. View Article : Google Scholar : PubMed/NCBI
42	Albiñana E, Luengo JG, Baraibar AM, Muñoz MD, Gandía L, Solís JM and Hernández-Guijo JM: Choline induces opposite changes in pyramidal neuron excitability and synaptic transmission through a nicotinic receptor-independent process in hippocampal slices. Pflugers Arch. 469:779–795. 2017. View Article : Google Scholar : PubMed/NCBI
43	Huang M, Felix AR, Kwon S, Lowe D, Wallace T, Santarelli L and Meltzer HY: The alpha-7 nicotinic receptor partial agonist/5-HT3 antagonist RG3487 enhances cortical and hippocampal dopamine and acetylcholine release. Psychopharmacology (Berl). 231:2199–2210. 2014. View Article : Google Scholar : PubMed/NCBI
44	Stoiljkovic M, Kelley C, Nagy D, Hurst R and Hajós M: Activation of α7 nicotinic acetylcholine receptors facilitates long-term potentiation at the hippocampal-prefrontal cortex synapses in vivo. Eur Neuropsychopharmacol. 26:2018–2023. 2016. View Article : Google Scholar : PubMed/NCBI
45	Lagostena L, Trocme-Thibierge C, Morain P and Cherubini E: The partial alpha7 nicotine acetylcholine receptor agonist S 24795 enhances long-term potentiation at CA3-CA1 synapses in the adult mouse hippocampus. Neuropharmacology. 54:676–685. 2008. View Article : Google Scholar : PubMed/NCBI
46	Inestrosa NC, Godoy JA, Vargas JY, Arrazola MS, Rios JA, Carvajal FJ, Serrano FG and Farias GG: Nicotine prevents synaptic impairment induced by amyloid-β oligomers through α7-nicotinic acetylcholine receptor activation. Neuromolecular Med. 15:549–569. 2013. View Article : Google Scholar : PubMed/NCBI
47	Counts SE, Nadeem M, Lad SP, Wuu J and Mufson EJ: Differential expression of synaptic proteins in the frontal and temporal cortex of elderly subjects with mild cognitive impairment. J Neuropathol Exp Neurol. 65:592–601. 2006. View Article : Google Scholar : PubMed/NCBI
48	Cao Y, Xiao Y, Ravid R and Guan ZZ: Changed clathrin regulatory proteins in the brains of Alzheimer's disease patients and animal models. J Alzheimers Dis. 22:329–342. 2010. View Article : Google Scholar : PubMed/NCBI
49	Carvalho C, Santos MS, Oliveira CR and Moreira PI: Alzheimer's disease and type 2 diabetes-related alterations in brain mitochondria, autophagy and synaptic markers. Biochim Biophys Acta. 1852:1665–1675. 2015. View Article : Google Scholar : PubMed/NCBI
50	Ferreira ST and Klein WL: The Aβ oligomer hypothesis for synapse failure and memory loss in Alzheimer's disease. Neurobiol Learn Mem. 96:529–543. 2011. View Article : Google Scholar : PubMed/NCBI
51	Tu S, Okamoto S, Lipton SA and Xu H: Oligomeric Aβ-induced synaptic dysfunction in Alzheimer's disease. Mol Neurodegener. 9:482014. View Article : Google Scholar : PubMed/NCBI
52	Wang S, Yu L, Yang H, Li C, Hui Z, Xu Y and Zhu X: Oridonin attenuates synaptic loss and cognitive deficits in an Aβ1-42-induced mouse model of Alzheimer's disease. PLoS One. 11:e01513972016. View Article : Google Scholar : PubMed/NCBI
53	Liu SJ, Yang C, Zhang Y, Su RY, Chen JL, Jiao MM, Chen HF, Zheng N, Luo S, Chen YB, et al: Neuroprotective effect of β-asarone against Alzheimer's disease: Regulation of synaptic plasticity by increased expression of SYP and GluR1. Drug Des Devel Ther. 10:1461–1469. 2016. View Article : Google Scholar : PubMed/NCBI
54	Chauhan NB, Lichtor T and Siegel GJ: Aging potentiates Abeta-induced depletion of SNAP-25 in mouse hippocampus. Brain Res. 982:219–227. 2003. View Article : Google Scholar : PubMed/NCBI
55	Qi XL, Nordberg A, Xiu J and Guan ZZ: The consequences of reducing expression of the alpha7 nicotinic receptor by RNA interference and of stimulating its activity with an alpha7 agonist in SH-SY5Y cells indicate that this receptor plays a neuroprotective role in connection with the pathogenesis of Alzheimer's disease. Neurochem Int. 51:377–383. 2007. View Article : Google Scholar : PubMed/NCBI