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Proc Natl Acad Sci U S A. 1996 Jun 25; 93(13): 6753–6758.

Amyloid beta-peptide disrupts carbachol-induced muscarinic cholinergic signal transduction in cortical neurons.


Cholinergic pathways serve important functions in learning and memory processes, and deficits in cholinergic transmission occur in Alzheimer disease (AD). A subset of muscarinic cholinergic receptors are linked to G-proteins that activate phospholipase C, resulting in the liberation of inositol trisphosphate and Ca2+ release from intracellular stores. We now report that amyloid beta-peptide (Abeta), which forms plaques in the brain in AD, impairs muscarinic receptor activation of G proteins in cultured rat cortical neurons. Exposure of rodent fetal cortical neurons to Abeta25-35 and Abeta1-40 resulted in a concentration and time-dependent attenuation of carbachol-induced GTPase activity without affecting muscarinic receptor ligand binding parameters. Downstream events in the signal transduction cascade were similarly attenuated by Abeta. Carbachol-induced accumulation of inositol phosphates (IP, IP2, IP3, and IP4) was decreased and calcium imaging studies revealed that carbachol-induced release of calcium was severely impaired in neurons pretreated with Abeta. Muscarinic cholinergic signal transduction was disrupted with subtoxic levels of exposure to AP. The effects of Abeta on carbachol-induced GTPase activity and calcium release were attenuated by antioxidants, implicating free radicals in the mechanism whereby Abeta induced uncoupling of muscarinic receptors. These data demonstrate that Abeta disrupts muscarinic receptor coupling to G proteins that mediate induction of phosphoinositide accumulation and calcium release, findings that implicate Abeta in the impairment of cholinergic transmission that occurs in AD.

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  • Bartus RT, Dean RL, 3rd, Beer B, Lippa AS. The cholinergic hypothesis of geriatric memory dysfunction. Science. 1982 Jul 30;217(4558):408–414. [PubMed]
  • Collerton D. Cholinergic function and intellectual decline in Alzheimer's disease. Neuroscience. 1986 Sep;19(1):1–28. [PubMed]
  • Exton JH. Role of G proteins in activation of phosphoinositide phospholipase C. Adv Second Messenger Phosphoprotein Res. 1993;28:65–72. [PubMed]
  • Smith CJ, Perry EK, Perry RH, Fairbairn AF, Birdsall NJ. Guanine nucleotide modulation of muscarinic cholinergic receptor binding in postmortem human brain--a preliminary study in Alzheimer's disease. Neurosci Lett. 1987 Nov 23;82(2):227–232. [PubMed]
  • Flynn DD, Weinstein DA, Mash DC. Loss of high-affinity agonist binding to M1 muscarinic receptors in Alzheimer's disease: implications for the failure of cholinergic replacement therapies. Ann Neurol. 1991 Mar;29(3):256–262. [PubMed]
  • Warpman U, Alafuzoff I, Nordberg A. Coupling of muscarinic receptors to GTP proteins in postmortem human brain--alterations in Alzheimer's disease. Neurosci Lett. 1993 Feb 5;150(1):39–43. [PubMed]
  • Bridges RS, Felicio LF, Pellerin LJ, Stuer AM, Mann PE. Prior parity reduces post-coital diurnal and nocturnal prolactin surges in rats. Life Sci. 1993;53(5):439–445. [PubMed]
  • Pearce BD, Potter LT. Coupling of m1 muscarinic receptors to G protein in Alzheimer disease. Alzheimer Dis Assoc Disord. 1991 Fall;5(3):163–172. [PubMed]
  • Cutler R, Joseph JA, Yamagami K, Villalobos-Molina R, Roth GS. Area specific alterations in muscarinic stimulated low Km GTPase activity in aging and Alzheimer's disease: implications for altered signal transduction. Brain Res. 1994 Nov 21;664(1-2):54–60. [PubMed]
  • De Keyser J, Ebinger G, Vauquelin G. D1-dopamine receptor abnormality in frontal cortex points to a functional alteration of cortical cell membranes in Alzheimer's disease. Arch Neurol. 1990 Jul;47(7):761–763. [PubMed]
  • Ohm TG, Bohl J, Lemmer B. Reduced basal and stimulated (isoprenaline, Gpp(NH)p, forskolin) adenylate cyclase activity in Alzheimer's disease correlated with histopathological changes. Brain Res. 1991 Feb 1;540(1-2):229–236. [PubMed]
  • Cowburn RF, O'Neill C, Ravid R, Alafuzoff I, Winblad B, Fowler CJ. Adenylyl cyclase activity in postmortem human brain: evidence of altered G protein mediation in Alzheimer's disease. J Neurochem. 1992 Apr;58(4):1409–1419. [PubMed]
  • Ross BM, McLaughlin M, Roberts M, Milligan G, McCulloch J, Knowler JT. Alterations in the activity of adenylate cyclase and high affinity GTPase in Alzheimer's disease. Brain Res. 1993 Sep 17;622(1-2):35–42. [PubMed]
  • Schnecko A, Witte K, Bohl J, Ohm T, Lemmer B. Adenylyl cyclase activity in Alzheimer's disease brain: stimulatory and inhibitory signal transduction pathways are differently affected. Brain Res. 1994 May 2;644(2):291–296. [PubMed]
  • Huang HM, Gibson GE. Altered beta-adrenergic receptor-stimulated cAMP formation in cultured skin fibroblasts from Alzheimer donors. J Biol Chem. 1993 Jul 15;268(20):14616–14621. [PubMed]
  • Selkoe DJ. Physiological production of the beta-amyloid protein and the mechanism of Alzheimer's disease. Trends Neurosci. 1993 Oct;16(10):403–409. [PubMed]
  • Yankner BA, Duffy LK, Kirschner DA. Neurotrophic and neurotoxic effects of amyloid beta protein: reversal by tachykinin neuropeptides. Science. 1990 Oct 12;250(4978):279–282. [PubMed]
  • Koh JY, Yang LL, Cotman CW. Beta-amyloid protein increases the vulnerability of cultured cortical neurons to excitotoxic damage. Brain Res. 1990 Nov 19;533(2):315–320. [PubMed]
  • Mattson MP, Cheng B, Davis D, Bryant K, Lieberburg I, Rydel RE. beta-Amyloid peptides destabilize calcium homeostasis and render human cortical neurons vulnerable to excitotoxicity. J Neurosci. 1992 Feb;12(2):376–389. [PubMed]
  • Mattson MP, Tomaselli KJ, Rydel RE. Calcium-destabilizing and neurodegenerative effects of aggregated beta-amyloid peptide are attenuated by basic FGF. Brain Res. 1993 Sep 3;621(1):35–49. [PubMed]
  • Hartmann H, Eckert A, Müller WE. beta-Amyloid protein amplifies calcium signalling in central neurons from the adult mouse. Biochem Biophys Res Commun. 1993 Aug 16;194(3):1216–1220. [PubMed]
  • Weiss JH, Pike CJ, Cotman CW. Ca2+ channel blockers attenuate beta-amyloid peptide toxicity to cortical neurons in culture. J Neurochem. 1994 Jan;62(1):372–375. [PubMed]
  • Goodman Y, Mattson MP. Secreted forms of beta-amyloid precursor protein protect hippocampal neurons against amyloid beta-peptide-induced oxidative injury. Exp Neurol. 1994 Jul;128(1):1–12. [PubMed]
  • Hensley K, Carney JM, Mattson MP, Aksenova M, Harris M, Wu JF, Floyd RA, Butterfield DA. A model for beta-amyloid aggregation and neurotoxicity based on free radical generation by the peptide: relevance to Alzheimer disease. Proc Natl Acad Sci U S A. 1994 Apr 12;91(8):3270–3274. [PMC free article] [PubMed]
  • Behl C, Davis JB, Lesley R, Schubert D. Hydrogen peroxide mediates amyloid beta protein toxicity. Cell. 1994 Jun 17;77(6):817–827. [PubMed]
  • Butterfield DA, Hensley K, Harris M, Mattson M, Carney J. beta-Amyloid peptide free radical fragments initiate synaptosomal lipoperoxidation in a sequence-specific fashion: implications to Alzheimer's disease. Biochem Biophys Res Commun. 1994 Apr 29;200(2):710–715. [PubMed]
  • Mark RJ, Hensley K, Butterfield DA, Mattson MP. Amyloid beta-peptide impairs ion-motive ATPase activities: evidence for a role in loss of neuronal Ca2+ homeostasis and cell death. J Neurosci. 1995 Sep;15(9):6239–6249. [PubMed]
  • Xie Z, Jack-Hays M, Wang Y, Periyasamy SM, Blanco G, Huang WH, Askari A. Different oxidant sensitivities of the alpha 1 and alpha 2 isoforms of Na+/K(+)-ATPase expressed in baculovirus-infected insect cells. Biochem Biophys Res Commun. 1995 Feb 6;207(1):155–159. [PubMed]
  • Rohn TT, Hinds TR, Vincenzi FF. Ion transport ATPases as targets for free radical damage. Protection by an aminosteroid of the Ca2+ pump ATPase and Na+/K+ pump ATPase of human red blood cell membranes. Biochem Pharmacol. 1993 Aug 3;46(3):525–534. [PubMed]
  • Villalobos-Molina R, Joseph JA, Rabin BM, Kandasamy SB, Dalton TK, Roth GS. Iron-56 irradiation diminishes muscarinic but not alpha 1-adrenergic-stimulated low-Km GTPase in rat brain. Radiat Res. 1994 Dec;140(3):382–386. [PubMed]
  • Mattson MP, Barger SW, Begley JG, Mark RJ. Calcium, free radicals, and excitotoxic neuronal death in primary cell culture. Methods Cell Biol. 1995;46:187–216. [PubMed]
  • Cassel D, Selinger Z. Catecholamine-stimulated GTPase activity in turkey erythrocyte membranes. Biochim Biophys Acta. 1976 Dec 8;452(2):538–551. [PubMed]
  • Ghodsi-Hovsepian S, Messer WS, Jr, Hoss W. Differential coupling between muscarinic receptors and G-proteins in regions of the rat brain. Biochem Pharmacol. 1990 Apr 15;39(8):1385–1391. [PubMed]
  • Yamagami K, Joseph JA, Roth GS. Decrement of muscarinic receptor-stimulated low-KM GTPase in striatum and hippocampus from the aged rat. Brain Res. 1992 Apr 3;576(2):327–331. [PubMed]
  • Berridge MJ, Downes CP, Hanley MR. Lithium amplifies agonist-dependent phosphatidylinositol responses in brain and salivary glands. Biochem J. 1982 Sep 15;206(3):587–595. [PMC free article] [PubMed]
  • Wu TW, Fung KP, Zeng LH, Wu J, Nakamura H. Propyl gallate as a hepatoprotector in vitro and in vivo. Biochem Pharmacol. 1994 Jul 19;48(2):419–422. [PubMed]
  • French JF, Thomas CE, Downs TR, Ohlweiler DF, Carr AA, Dage RC. Protective effects of a cyclic nitrone antioxidant in animal models of endotoxic shock and chronic bacteremia. Circ Shock. 1994 Jul;43(3):130–136. [PubMed]
  • Araujo DM, Lapchak PA, Robitaille Y, Gauthier S, Quirion R. Differential alteration of various cholinergic markers in cortical and subcortical regions of human brain in Alzheimer's disease. J Neurochem. 1988 Jun;50(6):1914–1923. [PubMed]
  • Flynn DD, Ferrari-DiLeo G, Mash DC, Levey AI. Differential regulation of molecular subtypes of muscarinic receptors in Alzheimer's disease. J Neurochem. 1995 Apr;64(4):1888–1891. [PubMed]
  • McLaughlin M, Ross BM, Milligan G, McCulloch J, Knowler JT. Robustness of G proteins in Alzheimer's disease: an immunoblot study. J Neurochem. 1991 Jul;57(1):9–14. [PubMed]
  • Greenwood AF, Powers RE, Jope RS. Phosphoinositide hydrolysis, G alpha q, phospholipase C, and protein kinase C in post mortem human brain: effects of post mortem interval, subject age, and Alzheimer's disease. Neuroscience. 1995 Nov;69(1):125–138. [PubMed]
  • Khatoon S, Grundke-Iqbal I, Iqbal K. Guanosine triphosphate binding to beta-subunit of tubulin in Alzheimer's disease brain: role of microtubule-associated protein tau. J Neurochem. 1995 Feb;64(2):777–787. [PubMed]
  • Roth GS, Joseph JA, Mason RP. Membrane alterations as causes of impaired signal transduction in Alzheimer's disease and aging. Trends Neurosci. 1995 May;18(5):203–206. [PubMed]
  • Müller WE, Koch S, Eckert A, Hartmann H, Scheuer K. beta-Amyloid peptide decreases membrane fluidity. Brain Res. 1995 Mar 13;674(1):133–136. [PubMed]
  • Williams LR. Oxidative stress, age-related neurodegeneration, and the potential for neurotrophic treatment. Cerebrovasc Brain Metab Rev. 1995 Spring;7(1):55–73. [PubMed]
  • Etcheberrigaray R, Ito E, Kim CS, Alkon DL. Soluble beta-amyloid induction of Alzheimer's phenotype for human fibroblast K+ channels. Science. 1994 Apr 8;264(5156):276–279. [PubMed]
  • Zhang C, Lambert MP, Bunch C, Barber K, Wade WS, Krafft GA, Klein WL. Focal adhesion kinase expressed by nerve cell lines shows increased tyrosine phosphorylation in response to Alzheimer's A beta peptide. J Biol Chem. 1994 Oct 14;269(41):25247–25250. [PubMed]
  • Nitsch RM, Slack BE, Wurtman RJ, Growdon JH. Release of Alzheimer amyloid precursor derivatives stimulated by activation of muscarinic acetylcholine receptors. Science. 1992 Oct 9;258(5080):304–307. [PubMed]
  • Buxbaum JD, Oishi M, Chen HI, Pinkas-Kramarski R, Jaffe EA, Gandy SE, Greengard P. Cholinergic agonists and interleukin 1 regulate processing and secretion of the Alzheimer beta/A4 amyloid protein precursor. Proc Natl Acad Sci U S A. 1992 Nov 1;89(21):10075–10078. [PMC free article] [PubMed]
  • Mattson MP, Cheng B, Culwell AR, Esch FS, Lieberburg I, Rydel RE. Evidence for excitoprotective and intraneuronal calcium-regulating roles for secreted forms of the beta-amyloid precursor protein. Neuron. 1993 Feb;10(2):243–254. [PubMed]
  • da Penha Berzaghi M, Cooper J, Castrén E, Zafra F, Sofroniew M, Thoenen H, Lindholm D. Cholinergic regulation of brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) but not neurotrophin-3 (NT-3) mRNA levels in the developing rat hippocampus. J Neurosci. 1993 Sep;13(9):3818–3826. [PubMed]
  • Knipper M, da Penha Berzaghi M, Blöchl A, Breer H, Thoenen H, Lindholm D. Positive feedback between acetylcholine and the neurotrophins nerve growth factor and brain-derived neurotrophic factor in the rat hippocampus. Eur J Neurosci. 1994 Apr 1;6(4):668–671. [PubMed]

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