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J Virol. Nov 1997; 71(11): 8821–8831.
PMCID: PMC192348

A hypothalamic neuronal cell line persistently infected with scrapie prions exhibits apoptosis.

Abstract

Neuronal death and vacuolation are characteristics of the CNS degeneration found in prion diseases. Relatively few cultured cell lines have been identified that can be persistently infected with scrapie prions, and none of these cells show cytopathologic changes reminiscent of prion neuropathology. The differentiated neuronal cell line GT1, established from gonadotropin hormone releasing-hormone neurons immortalized by genetically targeted tumorigenesis in transgenic mice (P. L. Mellon, JJ. Windle, P. C. Goldsmith, C. A. Padula, J. L. Roberts, and R. I. Weiner, Neuron 5:1-10, 1990), was examined for its ability to support prion formation. We found that GT1 cells could be persistently infected with mouse RML prions and that conditioned medium from infected cells could transfer prions to uninfected cells. In many but not all experiments, a subpopulation of cells showed reduced viability, morphological signs of neurodegeneration and vacuolation, and features of apoptosis. Subclones of GT1 cells that were stably transfected with the trk4 gene encoding the high-affinity nerve growth factor (NGF) receptor (GT1-trk) could also be persistently infected. NGF increased the viability of the scrapie-infected GT1-trk cells and reduced the morphological and biochemical signs of vacuolation and apoptosis. GT1 cells represent a novel system for studying the molecular mechanisms underlying prion infectivity and subsequent neurodegenerative changes.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Altman J. Programmed cell death: the paths to suicide. Trends Neurosci. 1992 Aug;15(8):278–280. [PubMed]
  • Arrigo AP, Suhan JP, Welch WJ. Dynamic changes in the structure and intracellular locale of the mammalian low-molecular-weight heat shock protein. Mol Cell Biol. 1988 Dec;8(12):5059–5071. [PMC free article] [PubMed]
  • Borchelt DR, Scott M, Taraboulos A, Stahl N, Prusiner SB. Scrapie and cellular prion proteins differ in their kinetics of synthesis and topology in cultured cells. J Cell Biol. 1990 Mar;110(3):743–752. [PMC free article] [PubMed]
  • Bracci-Laudiero L, Vigneti E, Iannicola C, Aloe L. NGF retards apoptosis in chick embryo bursal cell in vitro. Differentiation. 1993 Jun;53(2):61–66. [PubMed]
  • Brown DR, Herms J, Kretzschmar HA. Mouse cortical cells lacking cellular PrP survive in culture with a neurotoxic PrP fragment. Neuroreport. 1994 Oct 27;5(16):2057–2060. [PubMed]
  • Brown DR, Schmidt B, Kretzschmar HA. Role of microglia and host prion protein in neurotoxicity of a prion protein fragment. Nature. 1996 Mar 28;380(6572):345–347. [PubMed]
  • Butler DA, Scott MR, Bockman JM, Borchelt DR, Taraboulos A, Hsiao KK, Kingsbury DT, Prusiner SB. Scrapie-infected murine neuroblastoma cells produce protease-resistant prion proteins. J Virol. 1988 May;62(5):1558–1564. [PMC free article] [PubMed]
  • Carlson GA, Goodman PA, Lovett M, Taylor BA, Marshall ST, Peterson-Torchia M, Westaway D, Prusiner SB. Genetics and polymorphism of the mouse prion gene complex: control of scrapie incubation time. Mol Cell Biol. 1988 Dec;8(12):5528–5540. [PMC free article] [PubMed]
  • Caughey B, Race RE, Ernst D, Buchmeier MJ, Chesebro B. Prion protein biosynthesis in scrapie-infected and uninfected neuroblastoma cells. J Virol. 1989 Jan;63(1):175–181. [PMC free article] [PubMed]
  • Chazot G, Broussolle E, Lapras C l, Blättler T, Aguzzi A, Kopp N. New variant of Creutzfeldt-Jakob disease in a 26-year-old French man. Lancet. 1996 Apr 27;347(9009):1181–1181. [PubMed]
  • Clarke MC, Millson GC. Infection of a cell line of mouse L fibroblasts with scrapie agent. Nature. 1976 May 13;261(5556):144–145. [PubMed]
  • Clarke PG. Developmental cell death: morphological diversity and multiple mechanisms. Anat Embryol (Berl) 1990;181(3):195–213. [PubMed]
  • Edwards SN, Tolkovsky AM. Characterization of apoptosis in cultured rat sympathetic neurons after nerve growth factor withdrawal. J Cell Biol. 1994 Feb;124(4):537–546. [PMC free article] [PubMed]
  • Fairbairn DW, Carnahan KG, Thwaits RN, Grigsby RV, Holyoak GR, O'Neill KL. Detection of apoptosis induced DNA cleavage in scrapie-infected sheep brain. FEMS Microbiol Lett. 1994 Jan 15;115(2-3):341–346. [PubMed]
  • Forloni G, Angeretti N, Chiesa R, Monzani E, Salmona M, Bugiani O, Tagliavini F. Neurotoxicity of a prion protein fragment. Nature. 1993 Apr 8;362(6420):543–546. [PubMed]
  • Gaido ML, Cidlowski JA. Identification, purification, and characterization of a calcium-dependent endonuclease (NUC18) from apoptotic rat thymocytes. NUC18 is not histone H2B. J Biol Chem. 1991 Oct 5;266(28):18580–18585. [PubMed]
  • Gajdusek DC. Unconventional viruses and the origin and disappearance of kuru. Science. 1977 Sep 2;197(4307):943–960. [PubMed]
  • Hefti F. Nerve growth factor promotes survival of septal cholinergic neurons after fimbrial transections. J Neurosci. 1986 Aug;6(8):2155–2162. [PubMed]
  • Holtzman DM, Li Y, Chen K, Gage FH, Epstein CJ, Mobley WC. Nerve growth factor reverses neuronal atrophy in a Down syndrome model of age-related neurodegeneration. Neurology. 1993 Dec;43(12):2668–2673. [PubMed]
  • Holtzman DM, Li Y, Parada LF, Kinsman S, Chen CK, Valletta JS, Zhou J, Long JB, Mobley WC. p140trk mRNA marks NGF-responsive forebrain neurons: evidence that trk gene expression is induced by NGF. Neuron. 1992 Sep;9(3):465–478. [PubMed]
  • Kane DJ, Sarafian TA, Anton R, Hahn H, Gralla EB, Valentine JS, Ord T, Bredesen DE. Bcl-2 inhibition of neural death: decreased generation of reactive oxygen species. Science. 1993 Nov 19;262(5137):1274–1277. [PubMed]
  • Koh JY, Cotman CW. Programmed cell death: its possible contribution to neurotoxicity mediated by calcium channel antagonists. Brain Res. 1992 Aug 7;587(2):233–240. [PubMed]
  • Kovács J, Fellinger E, Kárpáti PA, Kovács AL, László L. The turnover of autophagic vacuoles: evaluation by quantitative electron microscopy. Biomed Biochim Acta. 1986;45(11-12):1543–1547. [PubMed]
  • Kristensson K, Feuerstein B, Taraboulos A, Hyun WC, Prusiner SB, DeArmond SJ. Scrapie prions alter receptor-mediated calcium responses in cultured cells. Neurology. 1993 Nov;43(11):2335–2341. [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Laszlo L, Lowe J, Self T, Kenward N, Landon M, McBride T, Farquhar C, McConnell I, Brown J, Hope J, et al. Lysosomes as key organelles in the pathogenesis of prion encephalopathies. J Pathol. 1992 Apr;166(4):333–341. [PubMed]
  • Loo DT, Copani A, Pike CJ, Whittemore ER, Walencewicz AJ, Cotman CW. Apoptosis is induced by beta-amyloid in cultured central nervous system neurons. Proc Natl Acad Sci U S A. 1993 Sep 1;90(17):7951–7955. [PMC free article] [PubMed]
  • Markovits P, Dormont D, Delpech B, Court L, Latarjet R. Essais de propagation in vitro de l'agent scrapie dans des cellules nerveuses de souris. C R Seances Acad Sci III. 1981 Nov 2;293(8):413–417. [PubMed]
  • Martin DP, Schmidt RE, DiStefano PS, Lowry OH, Carter JG, Johnson EM., Jr Inhibitors of protein synthesis and RNA synthesis prevent neuronal death caused by nerve growth factor deprivation. J Cell Biol. 1988 Mar;106(3):829–844. [PMC free article] [PubMed]
  • Martínez de la Escalera G, Choi AL, Weiner RI. Generation and synchronization of gonadotropin-releasing hormone (GnRH) pulses: intrinsic properties of the GT1-1 GnRH neuronal cell line. Proc Natl Acad Sci U S A. 1992 Mar 1;89(5):1852–1855. [PMC free article] [PubMed]
  • Mellon PL, Windle JJ, Goldsmith PC, Padula CA, Roberts JL, Weiner RI. Immortalization of hypothalamic GnRH neurons by genetically targeted tumorigenesis. Neuron. 1990 Jul;5(1):1–10. [PubMed]
  • Mobley WC. Nerve growth factor in Alzheimer's disease: to treat or not to treat? Neurobiol Aging. 1989 Sep-Oct;10(5):578–590. [PubMed]
  • Newell MK, Haughn LJ, Maroun CR, Julius MH. Death of mature T cells by separate ligation of CD4 and the T-cell receptor for antigen. Nature. 1990 Sep 20;347(6290):286–289. [PubMed]
  • Pan KM, Baldwin M, Nguyen J, Gasset M, Serban A, Groth D, Mehlhorn I, Huang Z, Fletterick RJ, Cohen FE, et al. Conversion of alpha-helices into beta-sheets features in the formation of the scrapie prion proteins. Proc Natl Acad Sci U S A. 1993 Dec 1;90(23):10962–10966. [PMC free article] [PubMed]
  • Pattison IH, Jones KM. Modification of a strain of mouse-adapted scrapie by passage through rats. Res Vet Sci. 1968 Sep;9(5):408–410. [PubMed]
  • Pittman RN, Mills JC, DiBenedetto AJ, Hynicka WP, Wang S. Neuronal cell death: searching for the smoking gun. Curr Opin Neurobiol. 1994 Feb;4(1):87–94. [PubMed]
  • Prusiner SB. Novel proteinaceous infectious particles cause scrapie. Science. 1982 Apr 9;216(4542):136–144. [PubMed]
  • Prusiner SB. Molecular biology of prion diseases. Science. 1991 Jun 14;252(5012):1515–1522. [PubMed]
  • Prusiner SB, McKinley MP, Bowman KA, Bolton DC, Bendheim PE, Groth DF, Glenner GG. Scrapie prions aggregate to form amyloid-like birefringent rods. Cell. 1983 Dec;35(2 Pt 1):349–358. [PubMed]
  • Race RE, Fadness LH, Chesebro B. Characterization of scrapie infection in mouse neuroblastoma cells. J Gen Virol. 1987 May;68(Pt 5):1391–1399. [PubMed]
  • Raff MC. Social controls on cell survival and cell death. Nature. 1992 Apr 2;356(6368):397–400. [PubMed]
  • Rubenstein R, Carp RI, Callahan SM. In vitro replication of scrapie agent in a neuronal model: infection of PC12 cells. J Gen Virol. 1984 Dec;65(Pt 12):2191–2198. [PubMed]
  • Rubenstein R, Deng H, Race RE, Ju W, Scalici CL, Papini MC, Kascsak RJ, Carp RI. Demonstration of scrapie strain diversity in infected PC12 cells. J Gen Virol. 1992 Nov;73(Pt 11):3027–3031. [PubMed]
  • Rubenstein R, Deng H, Scalici CL, Papini MC. Alterations in neurotransmitter-related enzyme activity in scrapie-infected PC12 cells. J Gen Virol. 1991 Jun;72(Pt 6):1279–1285. [PubMed]
  • Schägger H, von Jagow G. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem. 1987 Nov 1;166(2):368–379. [PubMed]
  • Schätzl HM, von der Helm K. V-sis gene (SSV) is expressed transiently and independently of v-gag (SSAV) after infection of fibroblasts with SSV/SSAV. Oncogene. 1989 Sep;4(9):1095–1101. [PubMed]
  • Schwanzel-Fukuda M, Pfaff DW. Origin of luteinizing hormone-releasing hormone neurons. Nature. 1989 Mar 9;338(6211):161–164. [PubMed]
  • Serban D, Taraboulos A, DeArmond SJ, Prusiner SB. Rapid detection of Creutzfeldt-Jakob disease and scrapie prion proteins. Neurology. 1990 Jan;40(1):110–117. [PubMed]
  • Shimohama S, Ogawa N, Tamura Y, Akaike A, Tsukahara T, Iwata H, Kimura J. Protective effect of nerve growth factor against glutamate-induced neurotoxicity in cultured cortical neurons. Brain Res. 1993 Dec 31;632(1-2):296–302. [PubMed]
  • Sparkes RS, Simon M, Cohn VH, Fournier RE, Lem J, Klisak I, Heinzmann C, Blatt C, Lucero M, Mohandas T, et al. Assignment of the human and mouse prion protein genes to homologous chromosomes. Proc Natl Acad Sci U S A. 1986 Oct;83(19):7358–7362. [PMC free article] [PubMed]
  • Stahl N, Borchelt DR, Hsiao K, Prusiner SB. Scrapie prion protein contains a phosphatidylinositol glycolipid. Cell. 1987 Oct 23;51(2):229–240. [PubMed]
  • Taraboulos A, Jendroska K, Serban D, Yang SL, DeArmond SJ, Prusiner SB. Regional mapping of prion proteins in brain. Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7620–7624. [PMC free article] [PubMed]
  • Taraboulos A, Raeber AJ, Borchelt DR, Serban D, Prusiner SB. Synthesis and trafficking of prion proteins in cultured cells. Mol Biol Cell. 1992 Aug;3(8):851–863. [PMC free article] [PubMed]
  • Taraboulos A, Rogers M, Borchelt DR, McKinley MP, Scott M, Serban D, Prusiner SB. Acquisition of protease resistance by prion proteins in scrapie-infected cells does not require asparagine-linked glycosylation. Proc Natl Acad Sci U S A. 1990 Nov;87(21):8262–8266. [PMC free article] [PubMed]
  • Taraboulos A, Serban D, Prusiner SB. Scrapie prion proteins accumulate in the cytoplasm of persistently infected cultured cells. J Cell Biol. 1990 Jun;110(6):2117–2132. [PMC free article] [PubMed]
  • Tatzelt J, Zuo J, Voellmy R, Scott M, Hartl U, Prusiner SB, Welch WJ. Scrapie prions selectively modify the stress response in neuroblastoma cells. Proc Natl Acad Sci U S A. 1995 Mar 28;92(7):2944–2948. [PMC free article] [PubMed]
  • Vaux DL, Haecker G, Strasser A. An evolutionary perspective on apoptosis. Cell. 1994 Mar 11;76(5):777–779. [PubMed]
  • Welch WJ, Suhan JP. Cellular and biochemical events in mammalian cells during and after recovery from physiological stress. J Cell Biol. 1986 Nov;103(5):2035–2052. [PMC free article] [PubMed]
  • Wetsel WC, Valença MM, Merchenthaler I, Liposits Z, López FJ, Weiner RI, Mellon PL, Negro-Vilar A. Intrinsic pulsatile secretory activity of immortalized luteinizing hormone-releasing hormone-secreting neurons. Proc Natl Acad Sci U S A. 1992 May 1;89(9):4149–4153. [PMC free article] [PubMed]
  • Will RG, Ironside JW, Zeidler M, Cousens SN, Estibeiro K, Alperovitch A, Poser S, Pocchiari M, Hofman A, Smith PG. A new variant of Creutzfeldt-Jakob disease in the UK. Lancet. 1996 Apr 6;347(9006):921–925. [PubMed]
  • Wong K, Qiu Y, Hyun W, Nixon R, VanCleff J, Sanchez-Salazar J, Prusiner SB, DeArmond SJ. Decreased receptor-mediated calcium response in prion-infected cells correlates with decreased membrane fluidity and IP3 release. Neurology. 1996 Sep;47(3):741–750. [PubMed]
  • Wyllie AH, Kerr JF, Currie AR. Cell death: the significance of apoptosis. Int Rev Cytol. 1980;68:251–306. [PubMed]
  • Zhou J, Holtzman DM, Weiner RI, Mobley WC. Expression of TrkA confers neuron-like responsiveness to nerve growth factor on an immortalized hypothalamic cell line. Proc Natl Acad Sci U S A. 1994 Apr 26;91(9):3824–3828. [PMC free article] [PubMed]

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