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. 1996 Aug 20;93(17):9223–9228. doi: 10.1073/pnas.93.17.9223

Expression and analysis of presenilin 1 in a human neuronal system: localization in cell bodies and dendrites.

D G Cook 1, J C Sung 1, T E Golde 1, K M Felsenstein 1, B S Wojczyk 1, R E Tanzi 1, J Q Trojanowski 1, V M Lee 1, R W Doms 1
PMCID: PMC38623  PMID: 8799182

Abstract

Mutations in the recently identified presenilin 1 gene on chromosome 14 cause early onset familial Alzheimer disease (FAD). Herein we describe the expression and analysis of the protein coded by presenilin 1 (PS1) in NT2N neurons, a human neuronal model system. PS1 was expressed using recombinant Semliki Forest virions and detected by introduced antigenic tags or antisera to PS1-derived peptides. Immunoprecipitation revealed two major PS1 bands of approximately 43 and 50 kDa, neither of which were N-glycosylated or O-glycosylated. Immunoreactive PS1 was detected in cell bodies and dendrites of NT2N neurons but not in axons or on the cell surface. PS1 was also detected in BHK cells, where it was also intracellular and colocalized with calnexin, a marker for the rough endoplasmic reticulum. A mutant form of PS1 linked to FAD did not differ from the wild-type protein at the light microscopic level. The model system described here will enable studies of the function of PS1 in human neurons and the role of mutant PS1 in FAD.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Alzheimer's Disease Collaborative Group The structure of the presenilin 1 (S182) gene and identification of six novel mutations in early onset AD families. Nat Genet. 1995 Oct;11(2):219–222. doi: 10.1038/ng1095-219. [DOI] [PubMed] [Google Scholar]
  2. Campion D., Flaman J. M., Brice A., Hannequin D., Dubois B., Martin C., Moreau V., Charbonnier F., Didierjean O., Tardieu S. Mutations of the presenilin I gene in families with early-onset Alzheimer's disease. Hum Mol Genet. 1995 Dec;4(12):2373–2377. doi: 10.1093/hmg/4.12.2373. [DOI] [PubMed] [Google Scholar]
  3. Citron M., Oltersdorf T., Haass C., McConlogue L., Hung A. Y., Seubert P., Vigo-Pelfrey C., Lieberburg I., Selkoe D. J. Mutation of the beta-amyloid precursor protein in familial Alzheimer's disease increases beta-protein production. Nature. 1992 Dec 17;360(6405):672–674. doi: 10.1038/360672a0. [DOI] [PubMed] [Google Scholar]
  4. Cook D. G., Lee V. M., Doms R. W. Expression of foreign proteins in a human neuronal system. Methods Cell Biol. 1994;43(Pt A):289–303. doi: 10.1016/s0091-679x(08)60609-3. [DOI] [PubMed] [Google Scholar]
  5. Copeland C. S., Doms R. W., Bolzau E. M., Webster R. G., Helenius A. Assembly of influenza hemagglutinin trimers and its role in intracellular transport. J Cell Biol. 1986 Oct;103(4):1179–1191. doi: 10.1083/jcb.103.4.1179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cork L. C., Masters C., Beyreuther K., Price D. L. Development of senile plaques. Relationships of neuronal abnormalities and amyloid deposits. Am J Pathol. 1990 Dec;137(6):1383–1392. [PMC free article] [PubMed] [Google Scholar]
  7. Crandall J. E., Fischer I. Developmental regulation of microtubule-associated protein 2 expression in regions of mouse brain. J Neurochem. 1989 Dec;53(6):1910–1917. doi: 10.1111/j.1471-4159.1989.tb09261.x. [DOI] [PubMed] [Google Scholar]
  8. Cras P., Kawai M., Lowery D., Gonzalez-DeWhitt P., Greenberg B., Perry G. Senile plaque neurites in Alzheimer disease accumulate amyloid precursor protein. Proc Natl Acad Sci U S A. 1991 Sep 1;88(17):7552–7556. doi: 10.1073/pnas.88.17.7552. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Doms R. W., Helenius A. Quaternary structure of influenza virus hemagglutinin after acid treatment. J Virol. 1986 Dec;60(3):833–839. doi: 10.1128/jvi.60.3.833-839.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Doms R. W. Oligomerization and protein transport. Methods Enzymol. 1990;191:841–854. doi: 10.1016/0076-6879(90)91051-7. [DOI] [PubMed] [Google Scholar]
  11. Games D., Adams D., Alessandrini R., Barbour R., Berthelette P., Blackwell C., Carr T., Clemens J., Donaldson T., Gillespie F. Alzheimer-type neuropathology in transgenic mice overexpressing V717F beta-amyloid precursor protein. Nature. 1995 Feb 9;373(6514):523–527. doi: 10.1038/373523a0. [DOI] [PubMed] [Google Scholar]
  12. Hammond C., Braakman I., Helenius A. Role of N-linked oligosaccharide recognition, glucose trimming, and calnexin in glycoprotein folding and quality control. Proc Natl Acad Sci U S A. 1994 Feb 1;91(3):913–917. doi: 10.1073/pnas.91.3.913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Jirgensons B. Conformational transitions of non-helical proteins effected by dodecyl sulfate. Circular dichroism of alpha1-acid glycoprotein, Bence Jones protein, carbonic anhydrase B, deoxyribonuclease, pepsinogen, and plasminogen. Biochim Biophys Acta. 1976 May 20;434(1):58–68. doi: 10.1016/0005-2795(76)90035-0. [DOI] [PubMed] [Google Scholar]
  14. Karlinsky H., Vaula G., Haines J. L., Ridgley J., Bergeron C., Mortilla M., Tupler R. G., Percy M. E., Robitaille Y., Noldy N. E. Molecular and prospective phenotypic characterization of a pedigree with familial Alzheimer's disease and a missense mutation in codon 717 of the beta-amyloid precursor protein gene. Neurology. 1992 Aug;42(8):1445–1453. doi: 10.1212/wnl.42.8.1445. [DOI] [PubMed] [Google Scholar]
  15. Kovacs D. M., Fausett H. J., Page K. J., Kim T. W., Moir R. D., Merriam D. E., Hollister R. D., Hallmark O. G., Mancini R., Felsenstein K. M. Alzheimer-associated presenilins 1 and 2: neuronal expression in brain and localization to intracellular membranes in mammalian cells. Nat Med. 1996 Feb;2(2):224–229. doi: 10.1038/nm0296-224. [DOI] [PubMed] [Google Scholar]
  16. L'Hernault S. W., Arduengo P. M. Mutation of a putative sperm membrane protein in Caenorhabditis elegans prevents sperm differentiation but not its associated meiotic divisions. J Cell Biol. 1992 Oct;119(1):55–68. doi: 10.1083/jcb.119.1.55. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Levitan D., Greenwald I. Facilitation of lin-12-mediated signalling by sel-12, a Caenorhabditis elegans S182 Alzheimer's disease gene. Nature. 1995 Sep 28;377(6547):351–354. doi: 10.1038/377351a0. [DOI] [PubMed] [Google Scholar]
  18. Levy-Lahad E., Wijsman E. M., Nemens E., Anderson L., Goddard K. A., Weber J. L., Bird T. D., Schellenberg G. D. A familial Alzheimer's disease locus on chromosome 1. Science. 1995 Aug 18;269(5226):970–973. doi: 10.1126/science.7638621. [DOI] [PubMed] [Google Scholar]
  19. Liljeström P., Garoff H. A new generation of animal cell expression vectors based on the Semliki Forest virus replicon. Biotechnology (N Y) 1991 Dec;9(12):1356–1361. doi: 10.1038/nbt1291-1356. [DOI] [PubMed] [Google Scholar]
  20. Minami Y., Weissman A. M., Samelson L. E., Klausner R. D. Building a multichain receptor: synthesis, degradation, and assembly of the T-cell antigen receptor. Proc Natl Acad Sci U S A. 1987 May;84(9):2688–2692. doi: 10.1073/pnas.84.9.2688. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Nilsson T., Warren G. Retention and retrieval in the endoplasmic reticulum and the Golgi apparatus. Curr Opin Cell Biol. 1994 Aug;6(4):517–521. doi: 10.1016/0955-0674(94)90070-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Pleasure S. J., Page C., Lee V. M. Pure, postmitotic, polarized human neurons derived from NTera 2 cells provide a system for expressing exogenous proteins in terminally differentiated neurons. J Neurosci. 1992 May;12(5):1802–1815. doi: 10.1523/JNEUROSCI.12-05-01802.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Rogaev E. I., Sherrington R., Rogaeva E. A., Levesque G., Ikeda M., Liang Y., Chi H., Lin C., Holman K., Tsuda T. Familial Alzheimer's disease in kindreds with missense mutations in a gene on chromosome 1 related to the Alzheimer's disease type 3 gene. Nature. 1995 Aug 31;376(6543):775–778. doi: 10.1038/376775a0. [DOI] [PubMed] [Google Scholar]
  24. Selkoe D. J. Normal and abnormal biology of the beta-amyloid precursor protein. Annu Rev Neurosci. 1994;17:489–517. doi: 10.1146/annurev.ne.17.030194.002421. [DOI] [PubMed] [Google Scholar]
  25. Sherrington R., Rogaev E. I., Liang Y., Rogaeva E. A., Levesque G., Ikeda M., Chi H., Lin C., Li G., Holman K. Cloning of a gene bearing missense mutations in early-onset familial Alzheimer's disease. Nature. 1995 Jun 29;375(6534):754–760. doi: 10.1038/375754a0. [DOI] [PubMed] [Google Scholar]
  26. Simons M., de Strooper B., Multhaup G., Tienari P. J., Dotti C. G., Beyreuther K. Amyloidogenic processing of the human amyloid precursor protein in primary cultures of rat hippocampal neurons. J Neurosci. 1996 Feb 1;16(3):899–908. doi: 10.1523/JNEUROSCI.16-03-00899.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sorbi S., Nacmias B., Forleo P., Piacentini S., Sherrington R., Rogaev E., St George Hyslop P., Amaducci L. Missense mutation of S182 gene in Italian families with early-onset Alzheimer's disease. Lancet. 1995 Aug 12;346(8972):439–440. doi: 10.1016/s0140-6736(95)92809-x. [DOI] [PubMed] [Google Scholar]
  28. Suzuki N., Cheung T. T., Cai X. D., Odaka A., Otvos L., Jr, Eckman C., Golde T. E., Younkin S. G. An increased percentage of long amyloid beta protein secreted by familial amyloid beta protein precursor (beta APP717) mutants. Science. 1994 May 27;264(5163):1336–1340. doi: 10.1126/science.8191290. [DOI] [PubMed] [Google Scholar]
  29. Tanahashi H., Mitsunaga Y., Takahashi K., Tasaki H., Watanabe S., Tabira T. Missense mutation of S182 gene in Japanese familial Alzheimer's disease. Lancet. 1995 Aug 12;346(8972):440–440. doi: 10.1016/s0140-6736(95)92810-3. [DOI] [PubMed] [Google Scholar]
  30. Tanzi R. E., Vaula G., Romano D. M., Mortilla M., Huang T. L., Tupler R. G., Wasco W., Hyman B. T., Haines J. L., Jenkins B. J. Assessment of amyloid beta-protein precursor gene mutations in a large set of familial and sporadic Alzheimer disease cases. Am J Hum Genet. 1992 Aug;51(2):273–282. [PMC free article] [PubMed] [Google Scholar]
  31. Tu P. H., Elder G., Lazzarini R. A., Nelson D., Trojanowski J. Q., Lee V. M. Overexpression of the human NFM subunit in transgenic mice modifies the level of endogenous NFL and the phosphorylation state of NFH subunits. J Cell Biol. 1995 Jun;129(6):1629–1640. doi: 10.1083/jcb.129.6.1629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Van Broeckhoven C. Presenilins and Alzheimer disease. Nat Genet. 1995 Nov;11(3):230–232. doi: 10.1038/ng1195-230. [DOI] [PubMed] [Google Scholar]
  33. Ward C. L., Kopito R. R. Intracellular turnover of cystic fibrosis transmembrane conductance regulator. Inefficient processing and rapid degradation of wild-type and mutant proteins. J Biol Chem. 1994 Oct 14;269(41):25710–25718. [PubMed] [Google Scholar]
  34. Wasco W., Pettingell W. P., Jondro P. D., Schmidt S. D., Gurubhagavatula S., Rodes L., DiBlasi T., Romano D. M., Guenette S. Y., Kovacs D. M. Familial Alzheimer's chromosome 14 mutations. Nat Med. 1995 Sep;1(9):848–848. doi: 10.1038/nm0995-848a. [DOI] [PubMed] [Google Scholar]
  35. Wojczyk B., Shakin-Eshleman S. H., Doms R. W., Xiang Z. Q., Ertl H. C., Wunner W. H., Spitalnik S. L. Stable secretion of a soluble, oligomeric form of rabies virus glycoprotein: influence of N-glycan processing on secretion. Biochemistry. 1995 Feb 28;34(8):2599–2609. doi: 10.1021/bi00008a026. [DOI] [PubMed] [Google Scholar]

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