Advertisement

Altered gene expression in fetal Down Syndrome brain as revealed by the gene hunting technique of subtractive hybridization

  • E. Kitzmueller
  • O. Labudova
  • H. Rink
  • N. Cairns
  • G. Lubec
Conference paper

Summary

Information on gene expression in brain of patients with Down Syndrome (DS, trisomy 21) is limited and molecular biological research is focussing on mapping and sequencing chromosome 21. The information on gene expression in DS available follows the current concept of a gene dosage effect due to a third copy of chromosome 21 claiming overexpression of genes encoded on this chromosome.

Based upon the availability of fetal brain and recent technology of gene hunting, we decided to use subtractive hybridization to evaluate differences in gene expression between DS and control brains.

Subtractive hybridization was applied on two fetal brains with DS and two age and sex matched controls, 23rd week of gestation, and mRNA steady state levels were evaluated generating a subtractive library. Subtracted sequences were identified by gene bank and assigned by alignments to individual genes.

We found a series of up-and downregulated sequences consisting of chromosomal transcripts, enzymes of intermediary metabolism, hormones, transporters / channels and transcription factors (TFs).

We show that trisomy 21 or aneuploidy leads to the deterioration of gene expression and the derangement of transcripts describes the impairment of transport, carriers, channels, signaling, known metabolic and hormone imbalances. The dys-coordinated expression of transcription factors including homeobox genes, POU-domain TFs, helix-loop-helix-motifs, LIM domain containing TFs, leucine zippers, forkhead genes, maybe of pathophysiological significance for abnormal brain development and wiring found in patients with DS. This is the first description of the concomitant expression of a large series of sequences indicating disruption of the concerted action of genes in this disorder.

Keywords

Down Syndrome Down Syndrome Patient Subtractive Library Down Syndrome Brain Gene Hunting 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Arai Y, Mizuguchi M, Ikeda K, Takashima S (1995) Developmental changes of apolipoprotein E immunoreactivity in Down syndrome brains. Brain Res Dev Brain Res 87: 228–232PubMedCrossRefGoogle Scholar
  2. Bauer D, Warthoe P, Rhode I, Struss M (1994) PCR methods and applications. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, pp 97–108Google Scholar
  3. Benvenisty N, Ornitz DM, Bennett GL, Sahagan BG, Kuo A, Cardiff RD, Leder P (1992) Brain tumours and lymphomas in transgenic mice that carry HTLV-I LTR/c-myc and Ig/tax genes. Oncogene 7: 2399–2405PubMedGoogle Scholar
  4. Bertioli DJ, Schlichter UH, Adams MJ, Burrows PR, Steinbiss HH, Antoniw JF (1995) An analysis of differential display shows a strong bias towards high copy number mRNAs. Nucl Acids Res 23: 4520–4523PubMedCrossRefGoogle Scholar
  5. Brodsky G, Barnes T, Bleskan J, Becker L, Cox M, Patterson D (1997) The human GARS-AIRS-GART gene encodes two proteins which are differentially expressed during human brain development and temporally overexpressed in cerebellum of individuals with Down syndrome. Hum Mol Genet 6: 2043–2050PubMedCrossRefGoogle Scholar
  6. Casciola-Rosen LA, Miller DK, Anhalt GJ, Rosen A (1994) Specific cleavage of the 70-kDa protein component of the U1 small nuclear ribonucleoprotein is a characteristic biochemical feature of apoptotic cell death. J Biol Chem 269: 30757–30760PubMedGoogle Scholar
  7. Casciola-Rosen L, Nicholson DW, Chong T, Rowan KR, Thornberry NA, Miller DK, Rosen A (1996) Apopain/CPP32 cleaves proteins that are essential for cellular repair: a fundamental principle of apoptotic death [see comments]. J Exp Med 183: 1957–1964PubMedCrossRefGoogle Scholar
  8. Casiano CA, Martin SJ, Green DR, Tan EM (1996) Selective cleavage of nuclear autoantigens during CD95 (Fas/APO-1)-mediated T cell apoptosis. J Exp Med 184: 765–770PubMedCrossRefGoogle Scholar
  9. Choi KS, Lim IK, Brady JN, Kim SJ (1998) ICE-like protease (caspase) is involved in transforming growth factor betal-mediated apoptosis in FaO rat hepatoma cell line. Hepatology 27: 415–421PubMedCrossRefGoogle Scholar
  10. Cowan EP, Alexander RK, Daniel S, Kashanchi F, Brady JN (1997) Induction of tumor necrosis factor alpha in human neuronal cells by extracellular human T-cell lymphotropic virus type 1 Tax. J Virol 71: 6982–6989PubMedGoogle Scholar
  11. Dammann T, Wohlleben W (1992) A metalloprotease gene from Streptomyces coelicolor “Muller” and its transcriptional activator, a member of the LysR family. Mol Microbiol 6: 2267–2278PubMedCrossRefGoogle Scholar
  12. de la Monte SM, Xu YY, Hutchins GM, Wands JR (1996) Developmental patterns of neuronal thread protein gene expression in Down syndrome. J Neurol Sci 135:118–125PubMedCrossRefGoogle Scholar
  13. Dix DJ, Allen JW, Collins BW, Mori C, Nakamura N, Poorman-Allen P, Goulding EH, Eddy EM (1996) Targeted gene disruption of Hsp70-2 results in failed meiosis, germ cell apoptosis, and male infertility. Proc Natl Acad Sci USA 93: 3264–3268PubMedCrossRefGoogle Scholar
  14. Drucker DJ (1998) Glucagon-like peptides. Diabetes 47: 159–169PubMedCrossRefGoogle Scholar
  15. Epstein CJ (1992) Down syndrome (Trisomy 21). In: Scriver CR, Beaudet AL, Sly WS, Valle D (eds) The metabolic and molecular basis of inherited disease. McGraw Hill, New YorkGoogle Scholar
  16. Epstein CJ (1995) Epilogue: toward the twenty-first century with Down syndrome-a personal view of how far we have come and of how far we can reasonably expect to go. Prog Clin Biol Res 393: 241–246PubMedGoogle Scholar
  17. Fang-Kircher S (1999) Increased steady state mRNA Levels of DNA-repair genes XRCC1, ERCC2 and ERCC3 in brain of patients with Down Syndrom. Life Sci 64: 1689–1699PubMedCrossRefGoogle Scholar
  18. Farr CJ, Easty DJ, Ragoussis J, Collignon J, Lovell-Badge R, Goodfellow PN (1993) Characterization and mapping of the human SOX4 gene. Mamm Genome 4: 577–584PubMedCrossRefGoogle Scholar
  19. Fu W, Shah SR, Jiang H, Hilt DC, Dave HP, Joshi JB (1997) Transactivation of proenkephalin gene by HTLV-1 tax1 protein in glial cells: involvement of Fos/Jun complex at an AP-1 element in the proenkephalin gene promoter. J Neurovirol 3:16–27PubMedCrossRefGoogle Scholar
  20. Gerbl-Rieger S, Peters J, Kellermann J, Lottspeich F, Baumeister W (1991) Nucleotide and derived amino acid sequences of the major porin of Comamonas acidovorans and comparison of porin primary structures. J Bacteriol 173: 2196–2205PubMedGoogle Scholar
  21. Gerbl-Rieger S, Engelhardt H, Peters J, Kehl M, Lottspeich F, Baumeister W (1992) Topology of the anion-selective porin Omp32 from Comamonas acidovorans. J Struct Biol 108: 14–24PubMedCrossRefGoogle Scholar
  22. Giaid A, Gibson SJ, Herrero MT, Gentleman S, Legon S, Yanagisawa M, Masaki T, Ibrahim NB, Roberts GW, Rossi ML, et al (1991) Topographical localisation of endothelin mRNA and peptide immunoreactivity in neurones of the human brain. Histochemistry 95: 303–314PubMedCrossRefGoogle Scholar
  23. Golubnitchaya-Labudova O, Horecka T, Kapalla M, Perecko D, Kutejova E, Lubec G (1998) Thioredoxin from Streptomyces aureofaciens controls coiling of plasmid DNA. Life Sci 62: 397–412PubMedCrossRefGoogle Scholar
  24. Goodison KL, Parhad IM, White CLd, Sima AA, Clark AW (1993) Neuronal and glial gene expression in neocortex of Down’s syndrome and Alzheimer’s disease. J Neuropathol Exp Neurol 52: 192–198PubMedCrossRefGoogle Scholar
  25. Greber-Platzer S, Schatzmann-Turhani D, Wollenek G, Lubec G (1999) Evidence against the current hypothesis of “gene dosage effects” of trisomy 21: ets-2, encoded on chromosome 21 ” is not overexpressed in hearts of patients with Down Syndrome. Biochem Biophys Res Commun 254: 395–399PubMedCrossRefGoogle Scholar
  26. Greidinger EL, Miller DK, Yamin TT, Casciola-Rosen L, Rosen A (1996) Sequential activation of three distinct ICE-like activities in Fas-ligated Jurkat cells. FEES Lett 390: 299–303CrossRefGoogle Scholar
  27. He L, Fox MH (1997) Variation of heat shock protein 70 through the cell cycle in HL-60 cells and its relationship to apoptosis. Exp Cell Res 232: 64–71PubMedCrossRefGoogle Scholar
  28. Hiraoka Y, Komatsu N, Sakai Y, Ogawa M, Shiozawa M, Aiso S (1997) XLS13A and XLS13B: SRY-related genes of Xenopus laevis. Gene 197: 65–71PubMedCrossRefGoogle Scholar
  29. Hirota K, Matsui M, Iwata S, Nishiyama A, Mori K, Yodoi J (1997) AP-1 transcriptional activity is regulated by a direct association between thioredoxin and Ref-1. Proc Natl Acad Sci USA 94: 3633–3638PubMedCrossRefGoogle Scholar
  30. Hohfeld J (1998) Regulation of the heat shock conjugate Hsc70 in the mammalian cell: the characterization of the anti-apoptotic protein BAG-1 provides novel insights. Biol Chem 379: 269–274PubMedGoogle Scholar
  31. Hori K, Katayama M, Sato N, Ishii K, Waga S, Yodoi J (1994) Neuroprotection by glial cells through adult T cell leukemia-derived factor/human thioredoxin (ADF/TRX). Brain Res 652: 304–310PubMedCrossRefGoogle Scholar
  32. Hyman BT, West HL, Rebeck GW, Lai F, Mann DM (1995) Neuropathological changes in Down’s syndrome hippocampal formation. Effect of age and apolipoprotein E genotype. Arch Neurol 52: 373–378PubMedCrossRefGoogle Scholar
  33. Kanda H, Kojima M, Miyamoto N, Ito M, Takamatsu N, Yamashita S, Shiba T (1998) Rainbow trout Sox24, a novel member of the Sox family, is a transcriptional regulator during oogenesis. Gene 211: 251–257PubMedCrossRefGoogle Scholar
  34. Khan IA, Luduena RF (1991) Possible regulation of the in vitro assembly of bovine brain tubulin by the bovine thioredoxin system. Biochim Biophys Acta 1076: 289–297PubMedCrossRefGoogle Scholar
  35. Kuhlbrodt K, Herbarth B, Sock E, Enderich J, Hermans-Borgmeyer I, Wegner M (1998) Cooperative function of POU proteins and SOX proteins in glial cells. J Biol Chem 273: 16050–16057PubMedCrossRefGoogle Scholar
  36. Kwak HJ, Jun CD, Pae HO, Yoo JC, Park YC, Choi BM, Na YG, Park RK, Chung HT, Chung HY, Park WY, Seo JS (1998) The role of inducible 70-kDa heat shock protein in cell cycle control, differentiation, and apoptotic cell death of the human myeloid leukemic HL-60 cells. Cell Immunol 187: 1–12PubMedCrossRefGoogle Scholar
  37. Labudova O, Lubec G (1998) cAmp upregulates the transposable element mys-1: a possible link between signaling and mobile Dna. Life Sci 62: 431–437PubMedCrossRefGoogle Scholar
  38. Labudova O, Fang-Kircher S, Cairns N, Moenkemann H, Yeghiazaryan K, Lubec G (1998) Brain vasopressin levels in Down syndrome and Alzheimer’s disease. Brain Res 806: 55–59PubMedCrossRefGoogle Scholar
  39. Labudova O, Kitzmueller E, Rink H, Cairns N, Lubec G (1999) Increased phosphoglycerate kinase in the brains of patients with Down’s syndrome but not with Alzheimer’s disease. Clin Sci (Colch) 96: 279–285CrossRefGoogle Scholar
  40. Lemere CA, Blusztajn JK, Yamaguchi H, Wisniewski T, Saido TC, Selkoe DJ (1996) Sequence of deposition of heterogeneous amyloid beta-peptides and APO E in Down syndrome: implications for initial events in amyloid plaque formation. Neurobiol Dis 3: 16–32PubMedCrossRefGoogle Scholar
  41. Lippoldt A, Padilla CA, Gerst H, Andbjer B, Richter E, Holmgren A, Fuxe K (1995) Localization of thioredoxin in the rat brain and functional implications. J Neurosci 15: 6747–6756PubMedGoogle Scholar
  42. Mailhos C, Howard MK, Latchman DS (1994) Heat shock proteins hsp90 and hsp70 protect neuronal cells from thermal stress but not from programmed cell death. J Neurochem 63: 1787–1795PubMedCrossRefGoogle Scholar
  43. Mann DM, Pickering-Brown SM, Siddons MA, Iwatsubo T, Ihara Y, Asami-Odaka A, Suzuki N (1995) The extent of amyloid deposition in brain in patients with Down’s syndrome does not depend upon the apolipoprotein E genotype. Neurosci Lett 196: 105–108PubMedCrossRefGoogle Scholar
  44. Marks A, D OH, Lei M, Percy ME, Becker LE (1996) Accumulation of S100 beta mRNA and protein in cerebellum during infancy in Down syndrome and control subjects. Brain Res Mol Brain Res 36: 343–348PubMedCrossRefGoogle Scholar
  45. Marsault R, Vigne P, Breittmayer JP, Freiin C (1990) Astrocytes are target cells for endothelins and sarafotoxin. J Neurochem 54: 2142–2144PubMedCrossRefGoogle Scholar
  46. Mavria G, Hall KT, Jones RA, Blair GE (1998) Transcriptional regulation of MHC class I gene expression in rat oligodendrocytes. Biochem J 330: 155–161PubMedGoogle Scholar
  47. Mentlein R, von Kolszynski M, Sprang R, Lucius R (1990) Proline-specific proteases in cultivated neuronal and glial cells. Brain Res 527: 159–162PubMedCrossRefGoogle Scholar
  48. Mesner PW, Epting CL, Hegarty JL, Green SH (1995) A timetable of events during programmed cell death induced by trophic factor withdrawal from neuronal PC12 cells. J Neurosci 15: 7357–7366PubMedGoogle Scholar
  49. Mori C, Nakamura N, Dix DJ, Fujioka M, Nakagawa S, Shiota K, Eddy EM (1997) Morphological analysis of germ cell apoptosis during postnatal testis development in normal and Hsp 70-2 knockout mice. Dev Dyn 208: 125–136PubMedCrossRefGoogle Scholar
  50. Mosser DD, Caron AW, Bourget L, Denis-Larose C, Massie B (1997) Role of the human heat shock protein hsp70 in protection against stress-induced apoptosis. Mol Cell Biol 17: 5317–5327PubMedGoogle Scholar
  51. Nyunoya H, Morita T, Sato T, Honma S, Tsujimoto A, Shimotohno K (1994) Cloning of a cDNA encoding a DNA-binding protein TAXREB302 that is specific for the tax-responsive enhancer of HTLV-I. Gene 148: 371–373PubMedCrossRefGoogle Scholar
  52. Oyama F, Cairns NJ, Shimada H, Oyama R, Titani K, Ihara Y (1994) Down’s syndrome: up-regulation of beta-amyloid protein precursor and tau mRNAs and their defective coordination. J Neurochem 62: 1062–1066PubMedCrossRefGoogle Scholar
  53. Pash J, Smithgall T, Bustin M (1991) Chromosomal protein HMG-14 is overexpressed in Down syndrome. Exp Cell Res 193: 232–235PubMedCrossRefGoogle Scholar
  54. Poteryaev DA, Zakharov IS, Balaban PM, Belyavsky AV (1998) A novel neuropeptide precursor gene is expressed in the terrestrial snail central nervous system by a group of neurons that control mating behavior. J Neurobiol 35: 183–197PubMedCrossRefGoogle Scholar
  55. Risser D, You ZB, Cairns N, Herrera-Marschitz M, Seidl R, Schneider C, Terenius L, Lubec G (1996) Endogenous opioids in frontal cortex of patients with Down syndrome. Neurosci Lett 203: 111–114PubMedCrossRefGoogle Scholar
  56. Saitoh M, Nishitoh H, Fujii M, Takeda K, Tobiume K, Sawada Y, Kawabata M, Miyazono K, Ichijo H (1998) Mammalian thioredoxin is a direct inhibitor of apoptosis signal-regulating kinase (ASK) 1. Embo J 17: 2596–2606PubMedCrossRefGoogle Scholar
  57. Scaife RM, Margolis RL (1997) The role of the PH domain and SH3 binding domains in dynamin function. Cell Signal 9: 395–401PubMedCrossRefGoogle Scholar
  58. Schilham MW, Oosterwegel MA, Moerer P, Ya J, de Boer PA, van de Wetering M, Verbeek S, Lamers WH, Kruisbeek AM, Cumano A, Clevers H (1996) Defects in cardiac outflow tract formation and pro-B-lymphocyte expansion in mice lacking Sox-4. Nature 380: 711–714PubMedCrossRefGoogle Scholar
  59. Schilham MW, Moerer P, Cumano A, Clevers HC (1997) Sox-4 facilitates thymocyte differentiation. Eur J Immunol 27: 1292–1295PubMedCrossRefGoogle Scholar
  60. Seidl R, Greber S, Schuller E, Bernert G, Cairns N, Lubec G (1997) Evidence against increased oxidative DNA-damage in Down syndrome. Neurosci Lett 235: 137–140PubMedCrossRefGoogle Scholar
  61. Seidl R, Fang-Kircher S, Bidmon B, Cairns N, Lubec G (1999) Apoptosis-associated proteins p53 and APO-1/Fas (CD95) in brains of adult patients with Down syndrome [In Process Citation]. Neurosci Lett 260: 9–12PubMedCrossRefGoogle Scholar
  62. Sompayrac L, Jane S, Burn TC, Tenen DG, Danna KJ (1995) Overcoming limitations of the mRNA differential display technique. Nucl Acids Res 23: 4738–4739PubMedCrossRefGoogle Scholar
  63. Sullivan AM, Morton AJ (1996) Endothelins induce Fos expression in neurons and glia in organotypic cultures of rat cerebellum. J Neurochem 67: 1409–1418PubMedCrossRefGoogle Scholar
  64. Tewari M, Beidler DR, Dixit VM (1995) CrmA-inhibitable cleavage of the 70-kDa protein component of the U1 small nuclear ribonucleoprotein during Fas-and tumor necrosis factor-induced apoptosis. J Biol Chem 270: 18738–18741PubMedCrossRefGoogle Scholar
  65. Torre E, McNiven MA, Urrutia R (1994) Dynamin 1 antisense oligonucleotide treatment prevents neurite formation in cultured hippocampal neurons. J Biol Chem 269: 32411–32417PubMedGoogle Scholar
  66. Tyrrell J, Cosgrave M, Hawi Z, McPherson J, C OB, McCalvert J, McLaughlin M, Lawlor B, Gill M (1998) A protective effect of apolipoprotein E e2 allele on dementia in Down’s syndrome. Biol Psychiatry 43: 397–400PubMedCrossRefGoogle Scholar
  67. Urrutia R, Henley JR, Cook T, McNiven MA (1997) The dynamins: redundant or distinct functions for an expanding family of related GTPases? Proc Natl Acad Sci USA 94: 377–384PubMedCrossRefGoogle Scholar
  68. Uwanogho D, Rex M, Cartwright EJ, Pearl G, Healy C, Scotting PJ, Sharpe PT (1995) Embryonic expression of the chicken Sox2, Sox3 and Soxll genes suggests an interactive role in neuronal development. Mech Dev 49: 23–36PubMedCrossRefGoogle Scholar
  69. van de Wetering M, Oosterwegel M, van Norren K, Clevers H, Farr CJ, Easty DJ, Ragoussis J, Collignon J, Lovell-Badge R, Goodfellow PN (1993) Sox-4, an Sry-like HMG box protein, is a transcriptional activator in lymphocytes Characterization and mapping of the human SOX4 gene. Embo J 12: 3847–3854PubMedGoogle Scholar
  70. Wei YQ, Zhao X, Kariya Y, Teshigawara K, Uchida A (1995) Inhibition of proliferation and induction of apoptosis by abrogation of heat-shock protein (HSP) 70 expression in tumor cells. Cancer Immunol Immunother 40: 73–78PubMedCrossRefGoogle Scholar
  71. Wyatt S, Mailhos C, Latchman DS (1996) Trigeminal ganglion neurons are protected by the heat shock proteins hsp70 and hsp90 from thermal stress but not from programmed cell death following nerve growth factor withdrawal. Brain Res Mol Brain Res 39: 52–56PubMedCrossRefGoogle Scholar
  72. Zhang W, Brooun A, McCandless J, Banda P, Alam M (1996) Signal transduction in the archaeon Halobacterium salinarium is processed through three subfamilies of 13 soluble and membrane-bound transducer proteins. Proc Natl Acad Sci USA 93:4649–4654PubMedCrossRefGoogle Scholar
  73. Zukowska-Grojec Z, Karwatowska-Prokopczuk E, Rose W, Rone J, Movafagh S, Ji H, Yeh Y, Chen WT, Kleinman HK, Grouzmann E, Grant DS (1998) Neuropeptide Y: a novel angiogenic factor from the sympathetic nerves and endothelium. Circ Res 83: 187–195PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Wien 1999

Authors and Affiliations

  • E. Kitzmueller
    • 1
  • O. Labudova
    • 1
  • H. Rink
    • 2
  • N. Cairns
    • 3
  • G. Lubec
    • 1
  1. 1.Department of PediatricsUniversity of ViennaViennaAustria
  2. 2.Department of RadiobiologyUniversity of BonnFederal Republic of Germany
  3. 3.Brain Bank, Institute of PsychiatryUniversity of LondonUK

Personalised recommendations