Top

Published in:

Open Access 01-01-2019 | Original Paper

The intact postsynaptic protein neurogranin is reduced in brain tissue from patients with familial and sporadic Alzheimer’s disease

Authors: Hlin Kvartsberg, Tammaryn Lashley, Christina E. Murray, Gunnar Brinkmalm, Nicholas C. Cullen, Kina Höglund, Henrik Zetterberg, Kaj Blennow, Erik Portelius

Published in: Acta Neuropathologica | Issue 1/2019

Abstract

Synaptic degeneration and neuronal loss are early events in Alzheimer’s disease (AD), occurring long before symptom onset, thus making synaptic biomarkers relevant for enabling early diagnosis. The postsynaptic protein neurogranin (Ng) is a cerebrospinal fluid (CSF) biomarker for AD, also in the prodromal phase. Here we tested the hypothesis that during AD neurodegeneration, processing of full-length Ng into endogenous peptides in the brain is increased. We characterized Ng in post-mortem brain tissue and investigated the levels of endogenous Ng peptides in relation to full-length protein in brain tissue of patients with sporadic (sAD) and familial Alzheimer’s disease (fAD), healthy controls and individuals who were cognitively unaffected but amyloid-positive (CU-AP) in two different brain regions. Brain tissue from parietal cortex [sAD (n = 10) and age-matched controls (n = 10)] and temporal cortex [sAD (n = 9), fAD (n = 10), CU-AP (n = 13) and controls (n = 9)] were included and all the samples were analyzed by three different methods. Using high-resolution mass spectrometry, 39 endogenous Ng peptides were identified while full-length Ng was found to be modified including disulfide bridges or glutathione. In sAD parietal cortex, the ratio of peptide-to-total full-length Ng was significantly increased for eight endogenous Ng peptides compared to controls. In the temporal cortex, several of the peptide-to-total full-length Ng ratios were increased in both sAD and fAD cases compared to controls and CU-AP. This finding was confirmed by western blot, which mainly detects full-length Ng, and enzyme-linked immunosorbent assay, most likely detecting a mix of peptides and full-length Ng. In addition, Ng was significantly associated with the degree of amyloid and tau pathology. These results suggest that processing of Ng into peptides is increased in AD brain tissue, which may reflect the ongoing synaptic degeneration, and which is also mirrored as increased levels of Ng peptides in CSF.

Available only for authorised users

Alzheimer’s Association (2016) 2016 Alzheimer’s disease facts and figures. Alzheimers Dement 12:459–509CrossRef

Aussedat B, Sagan S, Chassaing G, Bolbach G, Burlina F (2006) Quantification of the efficiency of cargo delivery by peptidic and pseudo-peptidic Trojan carriers using MALDI-TOF mass spectrometry. Biochim Biophys Acta 1758:375–383. https://doi.org/10.1016/j.bbamem.2006.01.012 CrossRefPubMed

Bahler M, Rhoads A (2002) Calmodulin signaling via the IQ motif. FEBS Lett 513:107–113CrossRefPubMed

Baudier J, Deloulme JC, Van Dorsselaer A, Black D, Matthes HW (1991) Purification and characterization of a brain-specific protein kinase C substrate, neurogranin (p17). Identification of a consensus amino acid sequence between neurogranin and neuromodulin (GAP43) that corresponds to the protein kinase C phosphorylation site and the calmodulin-binding domain. J Biol Chem 266:229–237PubMed

Becker B, Nazir FH, Brinkmalm G, Camporesi E, Kvartsberg H, Portelius E et al (2018) Alzheimer-associated cerebrospinal fluid fragments of neurogranin are generated by Calpain-1 and prolyl endopeptidase. Mol Neurodegener 13:47. https://doi.org/10.1186/s13024-018-0279-z CrossRefPubMedPubMedCentral

Bekris LM, Yu CE, Bird TD, Tsuang DW (2010) Genetics of Alzheimer disease. J Geriatr Psychiatry Neurol 23:213–227. https://doi.org/10.1177/0891988710383571 CrossRefPubMedPubMedCentral

Bennett DA, Schneider JA, Arvanitakis Z, Kelly JF, Aggarwal NT, Shah RC et al (2006) Neuropathology of older persons without cognitive impairment from two community-based studies. Neurology 66:1837–1844. https://doi.org/10.1212/01.wnl.0000219668.47116.e6 CrossRefPubMed

Blennow K, Bogdanovic N, Alafuzoff I, Ekman R, Davidsson P (1996) Synaptic pathology in Alzheimer’s disease: relation to severity of dementia, but not to senile plaques, neurofibrillary tangles, or the ApoE4 allele. J Neural Transm 103:603–618. https://doi.org/10.1007/BF01273157 (Vienna) CrossRefPubMed

Blennow K, de Leon MJ, Zetterberg H (2006) Alzheimer’s disease. Lancet 368:387–403. https://doi.org/10.1016/S0140-6736(06)69113-7 CrossRefPubMed

10.

Bogdanovic N, Davidsson P, Gottfries J, VIW B, Blennow K (2002) Regional and cellular distribution of synaptic proteins in the normal human brain. Brain Aging 2:18–30

11.

Braak H, Braak E (1991) Neuropathological stageing of Alzheimer-related changes. Acta Neuropathol 82:239–259CrossRefPubMed

12.

Brinkmalm G, Portelius E, Ohrfelt A, Mattsson N, Persson R, Gustavsson MK et al (2012) An online nano-LC-ESI-FTICR-MS method for comprehensive characterization of endogenous fragments from amyloid beta and amyloid precursor protein in human and cat cerebrospinal fluid. J Mass Spectrom 47:591–603. https://doi.org/10.1002/jms.2987 CrossRefPubMed

13.

Brouwers N, Sleegers K, Van Broeckhoven C (2008) Molecular genetics of Alzheimer’s disease: an update. Ann Med 40:562–583. https://doi.org/10.1080/07853890802186905 CrossRefPubMed

14.

Calkins MJ, Manczak M, Mao P, Shirendeb U, Reddy PH (2011) Impaired mitochondrial biogenesis, defective axonal transport of mitochondria, abnormal mitochondrial dynamics and synaptic degeneration in a mouse model of Alzheimer’s disease. Hum Mol Genet 20:4515–4529. https://doi.org/10.1093/hmg/ddr381 CrossRefPubMedPubMedCentral

15.

Davidsson P, Blennow K (1998) Neurochemical dissection of synaptic pathology in Alzheimer’s disease. Int Psychogeriatr 10:11–23CrossRefPubMed

16.

Davies CA, Mann DM, Sumpter PQ, Yates PO (1987) A quantitative morphometric analysis of the neuronal and synaptic content of the frontal and temporal cortex in patients with Alzheimer’s disease. J Neurol Sci 78:151–164CrossRefPubMed

17.

De Vos A, Jacobs D, Struyfs H, Fransen E, Andersson K, Portelius E et al (2015) C-terminal neurogranin is increased in cerebrospinal fluid but unchanged in plasma in Alzheimer’s disease. Alzheimers Dement 11:1461–1469. https://doi.org/10.1016/j.jalz.2015.05.012 CrossRefPubMed

18.

DeKosky ST, Scheff SW (1990) Synapse loss in frontal cortex biopsies in Alzheimer’s disease: correlation with cognitive severity. Ann Neurol 27:457–464. https://doi.org/10.1002/ana.410270502 CrossRefPubMed

19.

Di Domenico F, Cenini G, Sultana R, Perluigi M, Uberti D, Memo M et al (2009) Glutathionylation of the pro-apoptotic protein p53 in Alzheimer’s disease brain: implications for AD pathogenesis. Neurochem Res 34:727–733. https://doi.org/10.1007/s11064-009-9924-9 CrossRefPubMedPubMedCentral

20.

Diez-Guerra FJ (2010) Neurogranin, a link between calcium/calmodulin and protein kinase C signaling in synaptic plasticity. IUBMB Life 62:597–606. https://doi.org/10.1002/iub.357 CrossRefPubMed

21.

Dumurgier J, Schraen S, Gabelle A, Vercruysse O, Bombois S, Laplanche JL et al (2015) Cerebrospinal fluid amyloid-beta 42/40 ratio in clinical setting of memory centers: a multicentric study. Alzheimers Res Ther 7:30. https://doi.org/10.1186/s13195-015-0114-5 CrossRefPubMedPubMedCentral

22.

Hansson O, Zetterberg H, Buchhave P, Andreasson U, Londos E, Minthon L et al (2007) Prediction of Alzheimer’s disease using the CSF Abeta42/Abeta40 ratio in patients with mild cognitive impairment. Dement Geriatr Cogn Disord 23:316–320. https://doi.org/10.1159/000100926 CrossRefPubMed

23.

Hayashi Y (2009) Long-term potentiation: two pathways meet at neurogranin. EMBO J 28:2859–2860. https://doi.org/10.1038/emboj.2009.273 CrossRefPubMedPubMedCentral

24.

Hellwig K, Kvartsberg H, Portelius E, Andreasson U, Oberstein TJ, Lewczuk P et al (2015) Neurogranin and YKL-40: independent markers of synaptic degeneration and neuroinflammation in Alzheimer’s disease. Alzheimers Res Ther 7:74. https://doi.org/10.1186/s13195-015-0161-y CrossRefPubMedPubMedCentral

25.

Huang KP, Huang FL, Jager T, Li J, Reymann KG, Balschun D (2004) Neurogranin/RC3 enhances long-term potentiation and learning by promoting calcium-mediated signaling. J Neurosci 24:10660–10669. https://doi.org/10.1523/JNEUROSCI.2213-04.2004 CrossRefPubMedPubMedCentral

26.

Hyman BT, Phelps CH, Beach TG, Bigio EH, Cairns NJ, Carrillo MC et al (2012) National Institute on Aging-Alzheimer’s Association guidelines for the neuropathologic assessment of Alzheimer’s disease. Alzheimers Dement 8:1–13. https://doi.org/10.1016/j.jalz.2011.10.007 CrossRefPubMedPubMedCentral

27.

Jack CR Jr, Bennett DA, Blennow K, Carrillo MC, Dunn B, Haeberlein SB et al (2018) NIA-AA Research Framework: toward a biological definition of Alzheimer’s disease. Alzheimers Dement 14:535–562. https://doi.org/10.1016/j.jalz.2018.02.018 CrossRefPubMedPubMedCentral

28.

Janelidze S, Hertze J, Zetterberg H, Landqvist Waldo M, Santillo A, Blennow K et al (2016) Cerebrospinal fluid neurogranin and YKL-40 as biomarkers of Alzheimer’s disease. Ann Clin Transl Neurol 3:12–20. https://doi.org/10.1002/acn3.266 CrossRefPubMed

29.

Kamat PK, Kalani A, Rai S, Swarnkar S, Tota S, Nath C et al (2016) Mechanism of oxidative stress and synapse dysfunction in the pathogenesis of Alzheimer’s disease: understanding the therapeutics strategies. Mol Neurobiol 53:648–661. https://doi.org/10.1007/s12035-014-9053-6 CrossRefPubMed

30.

Kester MI, Teunissen CE, Crimmins DL, Herries EM, Ladenson JH, Scheltens P et al (2015) Neurogranin as a cerebrospinal fluid biomarker for synaptic loss in symptomatic Alzheimer disease. JAMA Neurol 72:1275–1280. https://doi.org/10.1001/jamaneurol.2015.1867 CrossRefPubMedPubMedCentral

31.

Knopman DS, Parisi JE, Salviati A, Floriach-Robert M, Boeve BF, Ivnik RJ et al (2003) Neuropathology of cognitively normal elderly. J Neuropathol Exp Neurol 62:1087–1095CrossRefPubMed

32.

Kvartsberg H, Duits FH, Ingelsson M, Andreasen N, Ohrfelt A, Andersson K et al (2015) Cerebrospinal fluid levels of the synaptic protein neurogranin correlates with cognitive decline in prodromal Alzheimer’s disease. Alzheimers Dement 11:1180–1190. https://doi.org/10.1016/j.jalz.2014.10.009 CrossRefPubMed

33.

Kvartsberg H, Portelius E, Andreasson U, Brinkmalm G, Hellwig K, Lelental N et al (2015) Characterization of the postsynaptic protein neurogranin in paired cerebrospinal fluid and plasma samples from Alzheimer’s disease patients and healthy controls. Alzheimers Res Ther 7:40. https://doi.org/10.1186/s13195-015-0124-3 CrossRefPubMedPubMedCentral

34.

Lewczuk P, Lelental N, Spitzer P, Maler JM, Kornhuber J (2015) Amyloid-beta 42/40 cerebrospinal fluid concentration ratio in the diagnostics of Alzheimer’s disease: validation of two novel assays. J Alzheimers Dis 43:183–191. https://doi.org/10.3233/JAD-140771 CrossRefPubMed

35.

Manczak M, Anekonda TS, Henson E, Park BS, Quinn J, Reddy PH (2006) Mitochondria are a direct site of A beta accumulation in Alzheimer’s disease neurons: implications for free radical generation and oxidative damage in disease progression. Hum Mol Genet 15:1437–1449. https://doi.org/10.1093/hmg/ddl066 CrossRefPubMed

36.

Masliah E, Hansen L, Albright T, Mallory M, Terry RD (1991) Immunoelectron microscopic study of synaptic pathology in Alzheimer’s disease. Acta Neuropathol 81:428–433CrossRefPubMed

37.

Masliah E, Mallory M, Alford M, DeTeresa R, Hansen LA, McKeel DW et al (2001) Altered expression of synaptic proteins occurs early during progression of Alzheimer’s disease. Neurology 56:127–129CrossRefPubMed

38.

Mattsson N, Insel PS, Palmqvist S, Portelius E, Zetterberg H, Weiner M et al (2016) Cerebrospinal fluid tau, neurogranin, and neurofilament light in Alzheimer’s disease. EMBO Mol Med 8:1184–1196. https://doi.org/10.15252/emmm.201606540 CrossRefPubMedPubMedCentral

39.

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 34:939–944CrossRefPubMed

40.

McKhann GM, Knopman DS, Chertkow H, Hyman BT, Jack CR Jr, Kawas CH et al (2011) The diagnosis of dementia due to Alzheimer’s disease: recommendations from the National Institute on Aging-Alzheimer’s Association workgroups on diagnostic guidelines for Alzheimer’s disease. Alzheimers Dement 7:263–269. https://doi.org/10.1016/j.jalz.2011.03.005 CrossRefPubMedPubMedCentral

41.

Mons N, Enderlin V, Jaffard R, Higueret P (2001) Selective age-related changes in the PKC-sensitive, calmodulin-binding protein, neurogranin, in the mouse brain. J Neurochem 79:859–867CrossRefPubMed

42.

Montine TJ, Phelps CH, Beach TG, Bigio EH, Cairns NJ, Dickson DW et al (2012) National Institute on Aging-Alzheimer’s Association guidelines for the neuropathologic assessment of Alzheimer’s disease: a practical approach. Acta Neuropathol 123:1–11. https://doi.org/10.1007/s00401-011-0910-3 CrossRefPubMed

43.

Pocernich CB, Butterfield DA (2012) Elevation of glutathione as a therapeutic strategy in Alzheimer disease. Biochim Biophys Acta 1822:625–630. https://doi.org/10.1016/j.bbadis.2011.10.003 CrossRefPubMed

44.

Portelius E, Lashley T, Westerlund A, Persson R, Fox NC, Blennow K et al (2015) Brain amyloid-beta fragment signatures in pathological ageing and Alzheimer’s disease by hybrid immunoprecipitation mass spectrometry. Neurodegener Dis 15:50–57. https://doi.org/10.1159/000369465 CrossRefPubMed

45.

Portelius E, Olsson B, Höglund K, Cullen NC, Kvartsberg H, Andreasson U et al (2018) Cerebrospinal fluid neurogranin concentration in neurodegeneration: relation to clinical phenotypes and neuropathology. Acta Neuropathol. https://doi.org/10.1007/s00401-018-1851-x CrossRefPubMedPubMedCentral

46.

Portelius E, Zetterberg H, Skillback T, Tornqvist U, Andreasson U, Trojanowski JQ et al (2015) Cerebrospinal fluid neurogranin: relation to cognition and neurodegeneration in Alzheimer’s disease. Brain 138:3373–3385. https://doi.org/10.1093/brain/awv267 CrossRefPubMedPubMedCentral

47.

Price JL, Davis PB, Morris JC, White DL (1991) The distribution of tangles, plaques and related immunohistochemical markers in healthy aging and Alzheimers-disease. Neurobiol Aging 12:295–312. https://doi.org/10.1016/0197-4580(91)90006-6 CrossRefPubMed

48.

Reddy PH, Beal MF (2008) Amyloid beta, mitochondrial dysfunction and synaptic damage: implications for cognitive decline in aging and Alzheimer’s disease. Trends Mol Med 14:45–53. https://doi.org/10.1016/j.molmed.2007.12.002 CrossRefPubMedPubMedCentral

49.

Reddy PH, Mani G, Park BS, Jacques J, Murdoch G, Whetsell W Jr et al (2005) Differential loss of synaptic proteins in Alzheimer’s disease: implications for synaptic dysfunction. J Alzheimers Dis 7:103–117 (discussion 173–180) CrossRefPubMed

50.

Rhoads AR, Friedberg F (1997) Sequence motifs for calmodulin recognition. FASEB J 11:331–340CrossRefPubMed

51.

Robinson JL, Molina-Porcel L, Corrada MM, Raible K, Lee EB, Lee VM et al (2014) Perforant path synaptic loss correlates with cognitive impairment and Alzheimer’s disease in the oldest-old. Brain 137:2578–2587. https://doi.org/10.1093/brain/awu190 CrossRefPubMedPubMedCentral

52.

Scheff SW, Price DA, Schmitt FA, DeKosky ST, Mufson EJ (2007) Synaptic alterations in CA1 in mild Alzheimer disease and mild cognitive impairment. Neurology 68:1501–1508. https://doi.org/10.1212/01.wnl.0000260698.46517.8f CrossRefPubMed

53.

Scheff SW, Price DA, Schmitt FA, Mufson EJ (2006) Hippocampal synaptic loss in early Alzheimer’s disease and mild cognitive impairment. Neurobiol Aging 27:1372–1384. https://doi.org/10.1016/j.neurobiolaging.2005.09.012 CrossRefPubMed

54.

Serrano-Pozo A, Frosch MP, Masliah E, Hyman BT (2011) Neuropathological alterations in Alzheimer disease. Cold Spring Harb Perspect Med 1:a006189. https://doi.org/10.1101/cshperspect.a006189 CrossRefPubMedPubMedCentral

55.

Shen J, Kelleher RJ 3rd (2007) The presenilin hypothesis of Alzheimer’s disease: evidence for a loss-of-function pathogenic mechanism. Proc Natl Acad Sci USA 104:403–409. https://doi.org/10.1073/pnas.0608332104 CrossRefPubMed

56.

Spies PE, Slats D, Sjogren JM, Kremer BP, Verhey FR, Rikkert MG et al (2010) The cerebrospinal fluid amyloid beta42/40 ratio in the differentiation of Alzheimer’s disease from non-Alzheimer’s dementia. Curr Alzheimer Res 7:470–476CrossRefPubMed

57.

Spires-Jones TL, Hyman BT (2014) The intersection of amyloid beta and tau at synapses in Alzheimer’s disease. Neuron 82:756–771. https://doi.org/10.1016/j.neuron.2014.05.004 CrossRefPubMedPubMedCentral

58.

Sze CI, Troncoso JC, Kawas C, Mouton P, Price DL, Martin LJ (1997) Loss of the presynaptic vesicle protein synaptophysin in hippocampus correlates with cognitive decline in Alzheimer disease. J Neuropathol Exp Neurol 56:933–944CrossRefPubMed

59.

Tarawneh R, D’Angelo G, Crimmins D, Herries E, Griest T, Fagan AM et al (2016) Diagnostic and prognostic utility of the synaptic marker neurogranin in Alzheimer disease. JAMA Neurol 73:561–571. https://doi.org/10.1001/jamaneurol.2016.0086 CrossRefPubMedPubMedCentral

60.

Thal DR, Rub U, Orantes M, Braak H (2002) Phases of A beta-deposition in the human brain and its relevance for the development of AD. Neurology 58:1791–1800CrossRefPubMed

61.

Thorsell A, Bjerke M, Gobom J, Brunhage E, Vanmechelen E, Andreasen N et al (2010) Neurogranin in cerebrospinal fluid as a marker of synaptic degeneration in Alzheimer’s disease. Brain Res 1362:13–22. https://doi.org/10.1016/j.brainres.2010.09.073 CrossRefPubMed

62.

Wiltfang J, Esselmann H, Bibl M, Hull M, Hampel H, Kessler H et al (2007) Amyloid beta peptide ratio 42/40 but not A beta 42 correlates with phospho-Tau in patients with low- and high-CSF A beta 40 load. J Neurochem 101:1053–1059. https://doi.org/10.1111/j.1471-4159.2006.04404.x CrossRefPubMed

63.

Xia ZG, Storm DR (2005) The role of calmodulin as a signal integrator for synaptic plasticity. Nat Rev Neurosci 6:267–276. https://doi.org/10.1038/nrn1647 CrossRefPubMed

64.

Zakharov VV, Mosevitsky MI (2001) Site-specific calcium-dependent proteolysis of neuronal protein GAP-43. Neurosci Res 39:447–453CrossRefPubMed

65.

Zhong L, Cherry T, Bies CE, Florence MA, Gerges NZ (2009) Neurogranin enhances synaptic strength through its interaction with calmodulin. EMBO J 28:3027–3039. https://doi.org/10.1038/emboj.2009.236 CrossRefPubMedPubMedCentral

Title: The intact postsynaptic protein neurogranin is reduced in brain tissue from patients with familial and sporadic Alzheimer’s disease
Authors: Hlin Kvartsberg
Tammaryn Lashley
Christina E. Murray
Gunnar Brinkmalm
Nicholas C. Cullen
Kina Höglund
Henrik Zetterberg
Kaj Blennow
Erik Portelius
Publication date: 01-01-2019
Publisher: Springer Berlin Heidelberg
Published in: Acta Neuropathologica / Issue 1/2019
Print ISSN: 0001-6322
Electronic ISSN: 1432-0533
DOI: https://doi.org/10.1007/s00401-018-1910-3

At a glance: The STEP trials

Springer Medicine

The intact postsynaptic protein neurogranin is reduced in brain tissue from patients with familial and sporadic Alzheimer’s disease

Abstract

At a glance: The STEP trials

Springer Medicine

Abstract

Please log in to get access to this content

Other articles of this Issue 1/2019

Rare variants in the neuronal ceroid lipofuscinosis gene MFSD8 are candidate risk factors for frontotemporal dementia

ETMR-like infantile cerebellar embryonal tumors in the extended morphologic spectrum of DICER1-related tumors

“Minimal change” multiple system atrophy with limbic-predominant α-synuclein pathology

Atypical parkinsonism of progressive supranuclear palsy–parkinsonism (PSP-P) phenotype with rare variants in FBXO7 and VPS35 genes associated with Lewy body pathology

Structure and evolution of double minutes in diagnosis and relapse brain tumors

The genetic landscape of gliomas arising after therapeutic radiation