Abstract
Increasing evidence links dysregulation of NR2B-containing N-methyl-d-aspartate receptor remodelling and trafficking to Alzheimer’s disease (AD). This theme offers the possibility that the GRIN2B gene, encoding this selective NR2B subunit, represents a potential molecular modulating factor for this disease. Based on this hypothesis, we carried out a mutation scanning of exons and flanking regions of GRIN2B in a well-characterized cohort of AD patients, recruited from Southern Italy. A “de novo” p.K1293R mutation, affecting a highly conserved residue of the protein in the C-terminal domain, was observed for the first time in a woman with familial AD, as the only genetic alteration of relevance. Moreover, an association study between the other detected sequence variants and AD was performed. In particular, the study was focused on five identified single nucleotide polymorphisms: rs7301328, rs1805482, rs3026160, rs1806191 and rs1806201, highlighting a significant contribution from the GRIN2B rs1806201 T allele towards disease susceptibility [adjusted odds ratio (OR) = 1.92, 95% confidence interval (CI) 1.40–2.63, p < 0.001, after correction for sex, age, and APOE ε4 genotype]. This was confirmed by haplotype analysis that identified a specific haplotype, carrying the rs1806201 T allele (CCCTC), over-represented in patients versus controls (adjusted OR = 6.03; p < 0.0001). Although the pathogenic role of the GRIN2B-K1293R mutation in AD is not clear, our data advocate that genetic variability in the GRIN2B gene, involved in synaptic functioning, might provide valuable insights into disease pathogenesis, continuing to attract significant attention in biomedical research on its genetic and functional role.
Similar content being viewed by others
References
Andreoli V, Trecroci F, La Russa A, Cittadella R, Liguori M, Spadafora P, Caracciolo M, Di Palma G, Colica C, Gambardella A, Quattrone A (2011) Presenilin enhancer-2 gene: identification of a novel promoter mutation in a patient with early-onset familial Alzheimer’s disease. Alzheimers Dement 7(6):574–578. doi:0.1016/.jalz.2011.2.010
Beste C, Baune BT, Domschke K, Falkenstein M, Konrad C (2010) Dissociable influences of NR2B-receptor related neural transmission on functions of distinct associative basal ganglia circuits. Neuroimage 52(1):309–315. doi:0.1016/.neuroimage.010.4.22
Bi H, Sze CI (2002) N-methyl-d-aspartate receptor subunit NR2A and NR2B messenger RNA levels are altered in the hippocampus and entorhinal cortex in Alzheimer’s disease. J Neurol Sci 200(1–2):11–18. doi:0.1016/S0022-510X
Cacabelos R (2008) Pharmacogenomics in Alzheimer’s disease. Methods Mol Biol 448:213–357. doi:10.1007/978-1-59745-205-2_10
Chamary JV, Hurst LD (2005) Evidence for selection on synonymous mutations affecting stability of mRNA secondary structure in mammals. Genome Biol 6(9):R75. doi:10.1186/gb-2005-6-9-r75
Chamary JV, Hurst LD (2009) The price of silent mutations. Sci Am 300(6):46–53
Chen C, Li X, Wang T, Wang HH, Fu Y, Zhang L, Xiao SF (2010) Association between NMDA receptor subunit 2b gene polymorphism and Alzheimer’s disease in Chinese Han population in Shanghai. Neurosci Bull 26(5):395–400. doi:10.1007/s12264-010-0729-2
Crum RM, Anthony JC, Bassett SS, Folstein M (1993) Population-based norms for the Mini- Mental State Examination by age and educational level. JAMA 269(18):2386–2391. doi:10.1001/jama.1993.03500180078038
Cruts M, van Duijin CM, Backhovens H, Van den Broeck M, Wehnert A, Serneels S, Sherrington R, Hutton M, Hardy J, St George-Hyslop PH, Hofman A, Van Broeckhoven C (1998) Estimation of the genetic contribution of presenilin-1 and -2 mutations in a population-based study of presenile Alzheimer disease. Hum Mol Genet 7(1):43–51. doi:10.1093/hmg/7.1.43
Cull-Candy SG, Leszkiewicz DN (2004) Role of distinct NMDA receptor subtypes at central synapses. Sci STKE 2004(255):re16
Danysz W, Parsons CG (2012) Alzheimer’s disease, β-amyloid, glutamate, NMDA receptors and memantine—searching for the connections. Br J Pharmacol 67(2):324–352. doi:10.1111/j.1476-5381.2012.02057
Darreh-Shori T, Siawesh M, Mousavi M, Andreasen N, Nordberg A (2012) Apolipoprotein ε4 modulates phenotype of butyrylcholinesterase in CSF of patients with Alzheimer’s disease. J Alzheimers Dis 28(2):443–458. doi:10.3233/JAD-2011-111088
Dorval KM, Wigg KG, Crosbie J, Tannock R, Kennedy JL, Ickowicz A, Pathare T, Malone M, Schachar R, Barr CL (2007) Association of the glutamate receptor subunit gene GRIN2B with attention-deficit/hyperactivity disorder. Genes Brain Behav 6(5):444–452. doi:10.1111/j.1601-183X.2006.00273.x
Endele S, Rosenberger G, Geider K, Popp B, Tamer C, Stefanova I, Milh M, Kortüm F, Fritsch A, Pientka FK, Hellenbroich Y, Kalscheuer VM, Kohlhase J, Moog U, Rappold G, Rauch A, Ropers HH, von Spiczak S, Tönnies H, Villeneuve N, Villard L, Zabel B, Zenker M, Laube B, Reis A, Wieczorek D, Van Maldergem L, Kutsche K (2010) Mutations in GRIN2A and GRIN2B encoding regulatory subunits of NMDA receptors cause variable neurodevelopmental phenotypes. Nat Genet 2(11):1021–1026. doi:10.1038/ng.677
Frueh FW, Noyer-Weidner M (2003) The use of denaturing high-performance liquid chromatography (DHPLC) for the analysis of genetic variations: impact for diagnostic and pharmacogenetics. Clin Chem Lab Med 41(4):452–461. doi:10.15/CCLM.2003.068
Goate AM, Chartier-Harlin MC, Mullan M, Brown J, Crawford F, Fidani L, Giuffra L, Haynes A, Irving N, James L et al (1991) Segregation of a missense mutation in the amyloid precursor protein gene with familial Alzheimer’s disease. Nature 349(6311):704–706. doi:10.1038/349704a0
Guerreiro RJ, Gustafson DR, Hardy J (2012) The genetic architecture of Alzheimer’s disease: beyond APP, PSENs and APOE. Neurobiol Aging 33(3):437–456. doi:10.1016/j.neurobiolaging.2010.03.025
Hawkins LM, Prybylowski K, Chang K, Moussan C, Stephenson FA, Wenthold RJ (2004) Export from the endoplasmic reticulum of assembled N-Methyl-d-aspartic Acid receptors is controlled by a motif in the C terminus of the NR2 subunit. J Biol Chem 279(28):28903–28910. doi:10.1074/jbc.M402599200
Hu NW, Ondrejcak T, Rowan MJ (2012) Glutamate receptors in preclinical research on Alzheimer’s disease: update on recent advances. Pharmacol Biochem Behav 100(4):855–862. doi:10.1016/j.pbb.2011.04.013
Jiang H, Jia J (2009) Association between NR2B subunit gene (GRIN2B) promoter polymorphisms and sporadic Alzheimer’s disease in the North Chinese population. Neurosci Lett 450(3):356–360. doi:10.1016/j.neulet.2008.10.075
Kutsuwada T, Sakimura K, Manabe T, Takayama C, Katakura N, Kushiya E, Natsume R, Watanabe M, Inoue Y, Yagi T, Aizawa S, Arakawa M, Takahashi T, Nakamura Y, Mori H, Mishina M (1996) Impairment of suckling response, trigeminal neuronal pattern formation, and hippocampal LTD in NMDA receptor epsilon 2 subunit mutant mice. Neuron 16(2):333–344. doi:10.1016/S0896-6273(00)80051-3
Lau CG, Zukin RS (2007) NMDA receptor trafficking in synaptic plasticity and neuropsychiatric disorders. Nat Rev Neurosci 8(6):413–426. doi:10.1038/nrn2153
Laube B, Hirai H, Sturgess M, Betz H, Kuhse J (1997) Molecular determinants of agonist discrimination by NMDA receptor subunits: analysis of the glutamate binding site on the NR2B subunit. Neuron 18(3):493–503. doi:10.1016/S0896-6273(00)81249-0
Lleó A, Castellví M, Blesa R, Oliva R (2002) Uncommon polymorphism in the presenilin genes in familial Alzheimer’s disease: not to be mistaken as a pathogenic mutation. Neurosci Lett 318(3):166–168. doi:10.1016/S0304-3940(01)02499-5
Luo L, Zhu Y, Xiong M (2012) A novel genome-information content-based statistic for genome-wide association analysis designed for next-generation sequencing data. J Comput Biol 19(6):731–744. doi:10.1089/cmb.2012.0035
McKhann G, Drachman D, Folstein M, Katzman R, Price D, Stadlan EM (1984) 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(7):939–944
Metzler M (2011) Mutations in NMDA receptors influence neurodevelopmental disorders causing epilepsy and intellectual disability. Clin Genet 79(3):219–220. doi:10.1111/j.1399-0004.2010.01610.x
Mony L, Kew JN, Gunthorpe MJ, Paoletti P (2009) Allosteric modulators of NR2B- containing NMDA receptors: molecular mechanisms and therapeutic potential. Br J Pharmacol 157(8):1301–1317. doi:10.1111/j.1476-5381.2009.00304.x
Monyer H, Sprengel R, Schoepfer R, Herb A, Higuchi M, Lomeli H, Burnashev N, Sakmann B, Seeburg PH (1992) Heteromeric NMDA receptors: molecular and functional distinction of subtypes. Science 256(5060):1217–1221. doi:10.1126/science.256.5060.1217
Olney JW, Wozniak DF, Farber NB (1997) Excitotoxic neurodegeneration in Alzheimer disease: new hypothesis and new therapeutic strategies. Arch Neurol 4(10):1234–1240. doi:10.1001/archneur.1997.00550220042012
Paoletti P (2011) Molecular basis of NMDA receptor functional diversity. Eur J Neurosci 33(8):1351–1365. doi:10.1111/j.1460-9568.2011.07628
Petralia RS (2012) Distribution of extrasynaptic NMDA receptors on neurons. Sci World J 2012:267120. doi:10.1100/2012/267120
Porsteinsson AP, Grossberg GT, Mintzer J, Olin JT, Memantine MEM-MD-12 Study Group (2008) Memantine treatment in patients) with mild to disease moderate Alzheimer’s already receiving a cholinesterase inhibitor: a randomized, double-blind, placebo-controlled trial. Curr Alzheimer Res 5(1):83–89. doi:10.2174/5672058783884576
Reisberg B, Doody R, Stöffler A, Schmitt F, Ferris S, Möbius HJ, Memantine Study Group (2003) Memantine in moderate to-severe Alzheimer’s disease. N Engl J Med 348(14):1333–1341. doi:10.1056/NEJMoa013128
Sadigh-Eteghad S, Talebi M, Farhoudi M (2012) Association of apolipoprotein E epsilon 4 allele with sporadic late onset Alzheimer’s disease: a meta-analysis. Neurosciences (Riyadh) 17(4):321–326
Santangelo RM, Acker TM, Zimmerman SS, Katzman BM, Strong KL, Traynelis SF, Liotta DC (2012) Novel NMDA receptor modulators: an update. Expert Opin Ther Pat 22(11):1337–1352. doi:10.1517/13543776.2012.728587
Schito AM, Pizzuti A, Di Maria E, Schenone A, Ratti A, Defferrari R, Bellone E, Mancardi GL, Ajmar F, Mandich P (1997) mRNA distribution in adult human brain of GRIN2B, a N-methyl-d-aspartate (NMDA) receptor subunit. Neurosci Lett 239(1):49–53. doi:0.1016/S0304-3940(97)00853-7
Seripa D, Matera MG, Franceschi M, Bizzarro A, Paris F, Cascavilla L, Rinaldi M, Panza F, Solfrizzi V, Daniele A, Masullo C, Dallapiccola B, Pilotto A (2008) Association analysis of GRIN2B, encoding N-methyl-d-aspartate receptor 2B subunit, and Alzheimer’s disease. Dement Geriatr Cogn Disord 25(3):287–292. doi:10.1159/000118634
Singh P, Doshi S, Spaethling JM, Hockenberry AJ, Patel TP, Geddes-Klein DM, Lynch DR, Meaney DF (2012) N methyl-D-aspartate receptor mechanosensitivity is governed by C-terminus of NR2B subunit. J Biol Chem 287(6):4348–4359. doi:10.1074/jbc.M111.253740
Snyder EM, Nong Y, Almeida CG, Paul S, Moran T, Choi EY, Nairn AC, Salter MW, Lombroso PJ, Gouras GK, Greengard P (2005) Regulation of NMDA receptor trafficking by amyloid-β. Nat Neurosci 8(8):1051–1058. doi:10.1038/nn1503
Sprengel R, Suchanek B, Amico C, Brusa R, Burnashev N, Rozov A, Hvalby O, Jensen V, Paulsen O, Andersen P, Kim JJ, Thompson RF, Sun W, Webster LC, Grant SGN, Eilers J, Konnerth A, Li J, McNamara JO, Seeburg PH (1998) Importance of the intracellular domain of NR2 subunits for NMDA receptor function in vivo. Cell 92(2):279–289. doi:10.1016/S0092-8674(00)80921-6
Stein JL, Hua X, Morra JH, Lee S, Hibar DP, Ho AJ, Leow AD, Toga AW, Sul JH, Kang HM, Eskin E, Saykin AJ, Shen L, Foroud T, Pankratz N, Huentelman MJ, Craig DW, Gerber JD, Allen AN, Corneveaux JJ, Stephan DA, Webster J, DeChairo BM, Potkin SG, Jack CR Jr, Weiner MW, Thompson PM, Alzheimer’s Disease Neuroimaging Initiative (2012) Genome-wide analysis reveals novel genes influencing temporal lobe structure with relevance to neurodegeneration in Alzheimer’s disease. Neuroimage 51(2):542–554. doi:10.1016/j.neuroimage.2010.02.068
Sze CI, Bi H, Kleinschmidt-DeMasters BK, Filley CM, Martin LJ (2001) NMDA receptor subunit NR1, NR2A, and NR2B proteins and their phosphorylation status are altered selectively in Alzheimer’s disease. J Neurol Sci 175(2):81–90. doi:10.1016/S0022-510X(00)00285-9
Tackenberg C, Grinschgl S, Trutzel A, Santuccione AC, Frey MC, Konietzko U, Grimm J, Brandt R, Nitsch RM (2013) NMDA receptor subunit composition determines beta-amyloid-induced neurodegeneration and synaptic loss. Cell Death Dis 25(4):e608. doi:10.1038/cddis.2013.129
Winblad B, Jones RW, Wirth Y, Stöffler A, Möbius HJ (2007) Memantine in moderate to severe Alzheimer’s disease: a meta-analysis of randomised clinical trials. Dement Geriatr Cogn Disord 24(1):20–27. doi:10.1159/000102568
Yang W, Zheng C, Song Q, Yang X, Qiu S, Liu C, Chen Z, Duan S, Luo J (2007) A three amino acid tail following the TM4 region of the N-methyl-d-aspartate receptor (NR)2 subunits is sufficient to overcome endoplasmic reticulum retention of NR1-1a subunit. J Biol Chem 282(12):9269–9278. doi:10.1074/jbc.M700050200
Acknowledgments
The authors thank the individuals with AD and their families for making this study possible. This study was partially supported by “Fondazione Carical” (Cosenza, Italy).
Conflict of interest
The authors have no conflicts of interest to disclose. Appropriate approval procedures were used concerning human subjects. There was no additional funding.
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
702_2013_1125_MOESM1_ESM.tif
Fig. 1 Molecular discovery and characterization of p.K1293R GRIN2B mutation by denaturing high-performance liquid chromatography (DHPLC) and direct DNA sequencing. The inset shows DHPLC analyses of a control (wild-type pattern with a1-peak trace) and the mutated sample (heterozygous state with three different peaks). Electropherogram of the patient demonstrates a lysine (Lys) to arginine (Arg) substitution at residue 1293
702_2013_1125_MOESM2_ESM.tif
Fig. 2a In silico protein analysis of the same GRIN2B amino acid substitutions by SIFT, PolyPhen, and SNAP programmes. Fig. 2b Evolutionary conservation of lysine 1293. Species, species-specific gene names, protein database accession numbers (www.ncbi.nlm.nih.gov/protein) and partial amino acids sequences are given. Orthologues residues identical to human NR2B K1293 are indicated in bold letters
Rights and permissions
About this article
Cite this article
Andreoli, V., De Marco, E.V., Trecroci, F. et al. Potential involvement of GRIN2B encoding the NMDA receptor subunit NR2B in the spectrum of Alzheimer’s disease. J Neural Transm 121, 533–542 (2014). https://doi.org/10.1007/s00702-013-1125-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00702-013-1125-7