Regulation of Amyloid Precursor Protein Secretion by Glutamate Receptors in Human Ntera 2 Neurons (NT2N)*

The amyloid precursor protein (APP) can be cleaved by a β-secretase to generate a β-amyloid peptide, which has been implicated in the pathogenesis of Alzheimer’s disease. However, APP can also be cleaved by an α-secretase to form a non-amyloidogenic secreted form of APP (APP-S). APP-S secretion can be physiologically regulated. This study examined the glutamatergic regulation of APP in the human neuronal Ntera 2 (NT2N) cell line. Metabotropic glutamate receptor subtypes 1α/β and 5α were identified in the NT2N neurons by reverse transcription-polymerase chain reaction. Stimulation of these phosphatidylinositol-linked receptors with glutamate or specific receptor agonists resulted in a dose- and time-dependent increase in the secretion of the amyloid precursor protein (APP-S), measured by the immunoprecipitation of APP-S from the medium of [35S]methionine-labeled NT2N neurons. The glutamate-induced APP-S secretion was maximal at 30 min and at a concentration of 1 mm glutamate. Glutamate-induced APP-S secretion required activation of phospholipase C, which resulted in inositol 1,4,5-trisphosphate production, as shown by the rapid glutamate-induced accumulation of inositol 1,4,5-trisphosphate. Glutamate also caused an increase in intracellular Ca2+. The protein kinase C activator phorbol 12-myristate 13-acetate, a phorbol ester, as well as 1-oleoyl-2-acetoyl-3-glycerol, a cell-permeable diacylglycerol analog, also stimulated APP-S secretion. These findings suggest that APP-S secretion from NT2N neurons can be regulated by the activation of phosphatidylinositol-linked metabotropic glutamate receptor signaling pathway.

[1]  D. Price,et al.  Evidence that beta-amyloid protein in Alzheimer's disease is not derived by normal processing. , 1990, Science.

[2]  John Hardy,et al.  Amyloid, the presenilins and Alzheimer's disease , 1997, Trends in Neurosciences.

[3]  D. Schoepp,et al.  Cloning and expression of a human metabotropic glutamate receptor 1 alpha: enhanced coupling on co-transfection with a glutamate transporter. , 1995, Molecular pharmacology.

[4]  J. Hardy,et al.  Amyloid deposition as the central event in the aetiology of Alzheimer's disease. , 1991, Trends in pharmacological sciences.

[5]  D. Selkoe,et al.  Enhanced Production and Oligomerization of the 42-residue Amyloid β-Protein by Chinese Hamster Ovary Cells Stably Expressing Mutant Presenilins* , 1997, The Journal of Biological Chemistry.

[6]  Burton S. Rosner,et al.  Neuropharmacology , 1958, Nature.

[7]  G. Westbrook.  Glutamate receptor update , 1994, Current Opinion in Neurobiology.

[8]  F. Young.  Biochemistry , 1955, The Indian Medical Gazette.

[9]  R. Wurtman,et al.  Metabotropic Glutamate Receptors Increase Amyloid Precursor Protein Processing in Astrocytes: Inhibition by Cyclic AMP , 1997, Journal of neurochemistry.

[10]  T. Oltersdorf,et al.  Cleavage of amyloid beta peptide during constitutive processing of its precursor. , 1990, Science.

[11]  V. Lee,et al.  Inducible expression of neuronal glutamate receptor channels in the NT2 human cell line. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[12]  V. Bigl,et al.  Glutamate-stimulated secretion of amyloid precursor protein from cortical rat brain slices , 1997, Neurochemistry International.

[13]  M. Mesulam,et al.  β-Amyloid and the pathogenesis of alzheimer's disease , 1991 .

[14]  J. Trojanowski,et al.  Human neurons derived from a teratocarcinoma cell line express solely the 695-amino acid amyloid precursor protein and produce intracellular beta-amyloid or A4 peptides. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[15]  N. Robakis,et al.  Intracellular Cyclic AMP Inhibits Constitutive and Phorbol Ester‐Stimulated Secretory Cleavage of Amyloid Precursor Protein , 1996, Journal of neurochemistry.

[16]  P. Chomczyński,et al.  Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. , 1987, Analytical biochemistry.

[17]  S. Younkin,et al.  Potentially amyloidogenic, carboxyl-terminal derivatives of the amyloid protein precursor. , 1992, Science.

[18]  L. Mucke,et al.  Alzheimer-type neuropathology in transgenic mice overexpressing V717F β-amyloid precursor protein , 1995, Nature.

[19]  D. Selkoe,et al.  Normal and abnormal biology of the beta-amyloid precursor protein. , 1994, Annual review of neuroscience.

[20]  E. Lightner,et al.  Mutation of the pancreatic islet inward rectifier Kir6.2 also leads to familial persistent hyperinsulinemic hypoglycemia of infancy. , 1996, Human molecular genetics.

[21]  D. Selkoe,et al.  The Role of APP Processing and Trafficking Pathways in the Formation of Amyloid β‐Protein a , 1996 .

[22]  Weiming Xia,et al.  Mutant presenilins of Alzheimer's disease increase production of 42-residue amyloid β-protein in both transfected cells and transgenic mice , 1997, Nature Medicine.

[23]  D. Sunter,et al.  New phenylglycine derivatives with potent and selective antagonist activity at presynaptic glutamate receptors in neonatal rat spinal cord , 1995, Neuropharmacology.

[24]  J. Trojanowski,et al.  Expression patterns of β‐amyloid precursor protein (β‐APP) in neural and nonneural human tissues from alzheimer's disease and control subjects , 1991 .

[25]  G. Lombardi,et al.  1-Aminoindan-1,5-dicarboxylic acid: a novel antagonist at phospholipase C-linked metabotropic glutamate receptors. , 1995, Journal of medicinal chemistry.

[26]  J. Mcculloch,et al.  Intracortical Glutamate Perfusionin VivoInduces Cellular Alterations in Specific Protein Kinase C Isoforms and Amyloid Precursor Protein , 1997, Experimental Neurology.

[27]  R. Wurtman,et al.  Amyloid precursor protein processing is stimulated by metabotropic glutamate receptors. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[28]  D. Selkoe,et al.  Processing of the amyloid protein precursor to potentially amyloidogenic derivatives. , 1992, Science.

[29]  S. Nakanishi,et al.  Agonist analysis of 2‐(carboxycyclopropyl)glycine isomers for cloned metabotropic glutamate receptor subtypes expressed in Chinese hamster ovary cells , 1992, British journal of pharmacology.

[30]  B. Wolf,et al.  Secretagogue-induced diacylglycerol accumulation in isolated pancreatic islets. Mass spectrometric characterization of the fatty acyl content indicates multiple mechanisms of generation. , 1989, Biochemistry.

[31]  V M Lee,et al.  Pure, postmitotic, polarized human neurons derived from NTera 2 cells provide a system for expressing exogenous proteins in terminally differentiated neurons , 1992, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[32]  B. Wolf,et al.  Muscarinic Regulation of Alzheimer's Disease Amyloid Precursor Protein Secretion and Amyloid β-Protein Production in Human Neuronal NT2N Cells (*) , 1995, The Journal of Biological Chemistry.

[33]  D. Price,et al.  Amyloidogenesis in Alzheimer's disease: basic biology and animal models , 1992, Current Opinion in Neurobiology.

[34]  S. Younkin,et al.  Release of excess amyloid beta protein from a mutant amyloid beta protein precursor. , 1993, Science.

[35]  P. Greengard,et al.  Metabolism of Alzheimer beta-amyloid precursor protein: regulation by protein kinase A in intact cells and in a cell-free system. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[36]  S. Younkin,et al.  Production of Amyloid β Protein from Normal Amyloid β‐Protein Precursor (βAPP) and the Mutated βAPPS Linked to Familial Alzheimer's Disease a , 1993 .

[37]  J. Growdon,et al.  Release of Alzheimer amyloid precursor derivatives stimulated by activation of muscarinic acetylcholine receptors. , 1992, Science.

[38]  N. Saijo,et al.  cDNA cloning of a short type of multidrug resistance protein homologue, SMRP, from a human lung cancer cell line. , 1997, Biochemical and biophysical research communications.

[39]  B. Wolf,et al.  Diacylglycerol hydrolysis to arachidonic acid is necessary for insulin secretion from isolated pancreatic islets: sequential actions of diacylglycerol and monoacylglycerol lipases. , 1994, Biochemistry.

[40]  Cha-Min Tang,et al.  Expression of Non‐NMDA Glutamate Receptor Channel Genes by Clonal Human Neurons , 1994, Journal of neurochemistry.

[41]  Allan I. Levey,et al.  Familial Alzheimer's Disease–Linked Presenilin 1 Variants Elevate Aβ1–42/1–40 Ratio In Vitro and In Vivo , 1996, Neuron.

[42]  R. Wurtman,et al.  Stimulation of amyloid precursor protein synthesis by adrenergic receptors coupled to cAMP formation. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[43]  R. Wurtman,et al.  Metabotropic glutamate receptor agonists increase release of soluble amyloid precursor protein derivatives from rat brain cortical and hippocampal slices. , 1997, The Journal of pharmacology and experimental therapeutics.

[44]  H. Sugiyama,et al.  Molecular cloning and the functional expression of two isoforms of human metabotropic glutamate receptor subtype 5. , 1994, Biochemical and biophysical research communications.