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A transgenic mouse model engineered to investigate human brain-derived neurotrophic factor in vivo

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Abstract

Brain-derived neurotrophic factor (BDNF) is an attractive component for the treatment of various neurodegenerative diseases such as Alzheimer’s or Parkinson’s disease. Innovative non-invasive therapeutic approaches involve appropriate pharmacological induction of endogenous BDNF synthesis in brain. A transgenic mouse model has been established to study human BDNF gene expression and permit the screening of compounds capable of stimulating its activity. A 145-kb yeast artificial chromosome carrying the human BDNF gene has been engineered to produce the transgene which contains the extended BDNF promoter and 3′ flanking regions and has integrated the enhanced green fluorescent protein (E-GFP) coding sequence in place of the BDNF coding exon. Five transgenic lines have been obtained through microinjection of the YAC into fertilized mouse oocytes. From the three lines expressing the transgene, one displays the specific pattern of BDNF expression. Faithful tissue-restricted transcription of BDNF 5′ exons and localization of the fluorescent reporter gene product in the expected brain subregions are reported. This line constitutes an exploitable system for investigating human BDNF gene regulation in vivo.

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Abbreviations

BDNF:

Brain-derived neurotrophic factor

YAC:

Yeast artificial chromosome

E-GFP:

Enhanced green fluorescent protein

STS:

Sequence-tagged site

kb:

Kilobase

nt:

Nucleotide

PCR:

Polymerase chain reaction

References

  • Barash I, Ilan N, Kari R et al (1996) Co-integration of beta-lactoglobulin/human serum albumin hybrid genes with the entire beta-lactoglobulin gene or the matrix attachment region element: repression of human serum albumin and beta-lactoglobulin expression in the mammary gland and dual regulation of the transgenes. Mol Reprod Dev 45:421–430

    Article  PubMed  CAS  Google Scholar 

  • Bishop JF, Mueller GP, Mouradian MM (1994) Alternate 5′ exons in the rat brain-derived neurotrophic factor gene: differential patterns of expression across brain regions. Brain Res Mol Brain Res 26:225–232

    Article  PubMed  CAS  Google Scholar 

  • Bode J, Stengert-Iber M, Kay V et al (1996) Scaffold/matrix-attached regions: topological switches with multiple regulatory functions. Crit Rev Eukaryot Gene Expr 6:115–138

    PubMed  CAS  Google Scholar 

  • Burke DT, Carle GF, Olson MV (1987) Cloning of large segments of exogenous DNA into yeast by means of artificial chromosome vectors. Science 236:806–812

    Article  PubMed  CAS  Google Scholar 

  • Carswell S (1993) The potential for treating neurodegenerative disorders with NGF-inducing compounds. Exp Neurol 124:36–42

    Article  PubMed  CAS  Google Scholar 

  • Conner JM, Lauterborn JC, Yan Q et al (1997) Distribution of brain-derived neurotrophic factor (BDNF) protein and mRNA in the normal adult rat CNS: evidence for anterograde axonal transport. J Neurosci 17:2295–2313

    PubMed  CAS  Google Scholar 

  • Connor B, Dragunow M (1998) The role of neuronal growth factors in neurodegenerative disorders of the human brain. Brain Res Brain Res Rev 27:1–39

    Article  PubMed  CAS  Google Scholar 

  • Cormack BP, Valdivia RH, Falkow S (1996) FACS-optimized mutants of the green fluorescent protein (GFP). Gene 173:33–38

    Article  PubMed  CAS  Google Scholar 

  • Cubitt AB, Heim R, Adams SR et al (1995) Understanding, improving and using green fluorescent proteins. Trends Biochem Sci 20:448–455

    Article  PubMed  CAS  Google Scholar 

  • da Penha Berzaghi M, Cooper J, Castren E et al (1993) 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 13:3818–3826

    PubMed  CAS  Google Scholar 

  • Davies AM, Wright EM (1995) Neurotrophic factors. Neurotrophin Autocrine loops. Curr Biol 5:723–726

    Article  PubMed  CAS  Google Scholar 

  • Duff K, McGuigan A, Huxley C et al (1994) Insertion of a pathogenic mutation into a yeast artificial chromosome containing the human amyloid precursor protein gene. Gene Ther 1:70–75

    PubMed  CAS  Google Scholar 

  • Eide FF, Lowenstein DH, Reichardt LF (1993) Neurotrophins and their receptors-current concepts and implications for neurologic disease. Exp Neurol 121:200–214

    Article  PubMed  CAS  Google Scholar 

  • Ferrer I, Goutan E, Marin C et al (2000) Brain-derived neurotrophic factor in Huntington disease. Brain Res 866:257–261

    Article  PubMed  CAS  Google Scholar 

  • Gervaix A, West D, Leoni LM et al (1997) A new reporter cell line to monitor HIV infection and drug susceptibility in vitro. Proc Natl Acad Sci USA 94:4653–4658

    Article  PubMed  CAS  Google Scholar 

  • Gimenez y Ribotta M, Revah F, Pradier L et al (1997) Prevention of motoneuron death by adenovirus-mediated neurotrophic factors. J Neurosci Res 48:281–285

    Article  PubMed  CAS  Google Scholar 

  • Girard C, Bemelmans AP, Dufour N et al (2005) Grafts of brain-derived neurotrophic factor and neurotrophin 3-transduced primate Schwann cells lead to functional recovery of the demyelinated mouse spinal cord. J Neurosci 25:7924–7933

    Article  PubMed  CAS  Google Scholar 

  • Godwin AR, Stadler HS, Nakamura K et al (1998) Detection of targeted GFP-Hox gene fusions during mouse embryogenesis. Proc Natl Acad Sci USA 95:13042–13047

    Article  PubMed  CAS  Google Scholar 

  • Guillemot F, Auffray C, Devignes MD (1999) Detailed transcript map of a 810-kb region at 11p14 involving identification of 10 novel human 3′ exons. Eur J Hum Genet 7:487–495

    Article  PubMed  CAS  Google Scholar 

  • Hallbook F, Lundin LG, Kullander K (1998) Lampetra fluviatilis neurotrophin homolog, descendant of a neurotrophin ancestor, discloses the early molecular evolution of neurotrophins in the vertebrate subphylum. J Neurosci 18:8700–8711

    PubMed  CAS  Google Scholar 

  • Hayashi M, Yamashita A, Shimizu K (1997) Somatostatin and brain-derived neurotrophic factor mRNA expression in the primate brain: decreased levels of mRNAs during aging. Brain Res 749:283–289

    Article  PubMed  CAS  Google Scholar 

  • Hayes VY, Towner MD, Isackson PJ (1997) Organization, sequence and functional analysis of a mouse BDNF promoter. Brain Res Mol Brain Res 45:189–198

    Article  PubMed  CAS  Google Scholar 

  • Hodgson JG, Smith DJ, McCutcheon K et al (1996) Human huntingtin derived from YAC transgenes compensates for loss of murine huntingtin by rescue of the embryonic lethal phenotype. Hum Mol Genet 5:1875–1885

    Article  PubMed  CAS  Google Scholar 

  • Hofer M, Pagliusi SR, Hohn A et al (1990) Regional distribution of brain-derived neurotrophic factor mRNA in the adult mouse brain. EMBO J 9:2459–2464

    PubMed  CAS  Google Scholar 

  • Hogan B, Beddington R, Costantini F et al (1994) In: Cold Spring Harbor Laboratory Press NY (ed) Manipulating the Mouse Embryo

  • Huxley C (1998) Exploring gene function: use of yeast artificial chromosome transgenesis. Methods 14:199–210

    Article  PubMed  CAS  Google Scholar 

  • Ikawa M, Kominami K, Yoshimura Y et al (1995) A rapid and non-invasive selection of transgenic embryos before implantation using green fluorescent protein (GFP). FEBS Lett 375:125–128

    Article  PubMed  CAS  Google Scholar 

  • Knusel B, Gao H (1996) Neurotrophins and Alzheimer’s disease: beyond the cholinergic neurons. Life Sci 58:2019–2027

    Article  PubMed  CAS  Google Scholar 

  • Lamb BT, Gearhart JD (1995) YAC transgenics and the study of genetics and human disease. Curr Opin Genet Dev 5:342–348

    Article  PubMed  CAS  Google Scholar 

  • Levi-Montalcini R (1987) The nerve growth factor: thirty-five years later. EMBO J 6:1145–1154

    PubMed  CAS  Google Scholar 

  • Levivier M, Przedborski S, Bencsics C et al (1995) Intrastriatal implantation of fibroblasts genetically engineered to produce brain-derived neurotrophic factor prevents degeneration of dopaminergic neurons in a rat model of Parkinson’s disease. J Neurosci 15:7810–7820

    PubMed  CAS  Google Scholar 

  • Liu QR, Walther D, Drgon T et al (2005) Human brain derived neurotrophic factor (BDNF) genes, splicing patterns, and assessments of associations with substance abuse and Parkinson’s Disease. Am J Med Genet B Neuropsychiatr Genet 134:93–103

    PubMed  Google Scholar 

  • Liu QR, Lu L, Zhu XG et al (2006) Rodent BDNF genes, novel promoters, novel splice variants, and regulation by cocaine. Brain Res 1067:1–12

    Article  PubMed  CAS  Google Scholar 

  • Ma YL, Wang HL, Wu HC et al (1998) Brain-derived neurotrophic factor antisense oligonucleotide impairs memory retention and inhibits long-term potentiation in rats. Neuroscience 82:957–967

    Article  PubMed  CAS  Google Scholar 

  • Manson AL, Trezise AE, MacVinish LJ et al (1997) Complementation of null CF mice with a human CFTR YAC transgene. EMBO J 16:4238–4249

    Article  PubMed  CAS  Google Scholar 

  • Martinez-Serrano A, Hantzopoulos PA, Bjorklund A (1996) Ex vivo gene transfer of brain-derived neurotrophic factor to the intact rat forebrain: neurotrophic effects on cholinergic neurons. Eur J Neurosci 8:727–735

    Article  PubMed  CAS  Google Scholar 

  • Mattson MP, Maudsley S, Martin B (2004) BDNF and 5-HT: a dynamic duo in age-related neuronal plasticity and neurodegenerative disorders. Trends Neurosci 27:589–594

    Article  PubMed  CAS  Google Scholar 

  • McAllister AK, Katz LC, Lo DC (1999) Neurotrophins and synaptic plasticity. Annu Rev Neurosci 22:295–318

    Article  PubMed  CAS  Google Scholar 

  • McKnight RA, Shamay A, Sankaran L et al (1992) Matrix-attachment regions can impart position-independent regulation of a tissue-specific gene in transgenic mice. Proc Natl Acad Sci USA 89:6943–6947

    Article  PubMed  CAS  Google Scholar 

  • Menei P, Montero-Menei C, Whittemore SR et al (1998) Schwann cells genetically modified to secrete human BDNF promote enhanced axonal regrowth across transected adult rat spinal cord. Eur J Neurosci 10:607–621

    Article  PubMed  CAS  Google Scholar 

  • Metsis M, Timmusk T, Arenas E et al (1993) Differential usage of multiple brain-derived neurotrophic factor promoters in the rat brain following neuronal activation. Proc Natl Acad Sci USA 90:8802–8806

    Article  PubMed  CAS  Google Scholar 

  • Misteli T, Spector DL (1997) Applications of the green fluorescent protein in cell biology and biotechnology. Nat Biotechnol 15:961–964

    Article  PubMed  CAS  Google Scholar 

  • Murer MG, Boissiere F, Yan Q et al (1999) An immunohistochemical study of the distribution of brain-derived neurotrophic factor in the adult human brain, with particular reference to Alzheimer’s disease. Neuroscience 88:1015–1032

    Article  PubMed  CAS  Google Scholar 

  • Murray KD, Gall CM, Jones EG et al (1994) Differential regulation of brain-derived neurotrophic factor and type II calcium/calmodulin-dependent protein kinase messenger RNA expression in Alzheimer’s disease. Neuroscience 60:37–48

    Article  PubMed  CAS  Google Scholar 

  • Namciu SJ, Blochlinger KB, Fournier RE (1998) Human matrix attachment regions insulate transgene expression from chromosomal position effects in Drosophila melanogaster. Mol Cell Biol 18:2382–2391

    PubMed  CAS  Google Scholar 

  • Neznanov N, Kohwi-Shigematsu T, Oshima RG (1996) Contrasting effects of the SATB1 core nuclear matrix attachment region and flanking sequences of the keratin 18 gene in transgenic mice. Mol Biol Cell 7:541–552

    PubMed  CAS  Google Scholar 

  • Nielsen LB, McCormick SP, Pierotti V et al (1997) Human apolipoprotein B transgenic mice generated with 207 and 145-kilobasepair bacterial artificial chromosomes. Evidence that a distant 5′-element confers appropriate transgene expression in the intestine. J Biol Chem 272:29752–29758

    Article  PubMed  CAS  Google Scholar 

  • Nonomura T, Nishio C, Lindsay RM et al (1995) Cultured basal forebrain cholinergic neurons from postnatal rats show both overlapping and non-overlapping responses to the neurotrophins. Brain Res 683:129–139

    Article  PubMed  CAS  Google Scholar 

  • Patterson SL, Abel T, Deuel TA et al (1996) Recombinant BDNF rescues deficits in basal synaptic transmission and hippocampal LTP in BDNF knockout mice. Neuron 16:1137–1145

    Article  PubMed  CAS  Google Scholar 

  • Phillips HS, Hains JM, Armanini M et al (1991) BDNF mRNA is decreased in the hippocampus of individuals with Alzheimer’s disease. Neuron 7:695–702

    Article  PubMed  CAS  Google Scholar 

  • van den Pol AN, Ghosh PK (1998) Selective neuronal expression of green fluorescent protein with cytomegalovirus promoter reveals entire neuronal arbor in transgenic mice. J Neurosci 18:10640–10651

    PubMed  Google Scholar 

  • Rosier MF, Goguel AF, Martin A et al (1994) A 1.7-Mb YAC contig around the human BDNF gene (11p13): integration of the physical, genetic, and cytogenetic maps in relation to WAGR syndrome. Genomics 24:69–77

    Article  PubMed  CAS  Google Scholar 

  • Saporito MS, Wilcox HM, Hartpence KC et al (1993) Pharmacological induction of nerve growth factor mRNA in adult rat brain. Exp Neurol 123:295–302

    Article  PubMed  CAS  Google Scholar 

  • Sarsero JP, Li L, Holloway TP et al (2004) Human BAC-mediated rescue of the Friedreich ataxia knockout mutation in transgenic mice. Mamm Genome 15:370–382

    Article  PubMed  CAS  Google Scholar 

  • Schedl A, Ross A, Lee M et al (1996) Influence of PAX6 gene dosage on development: overexpression causes severe eye abnormalities. Cell 86:71–82

    Article  PubMed  CAS  Google Scholar 

  • Semkova I, Krieglstein J (1999) Neuroprotection mediated via neurotrophic factors and induction of neurotrophic factors. Brain Res Brain Res Rev 30:176–188

    Article  PubMed  CAS  Google Scholar 

  • Singh M, Meyer EM, Simpkins JW (1995) The effect of ovariectomy and estradiol replacement on brain-derived neurotrophic factor messenger ribonucleic acid expression in cortical and hippocampal brain regions of female Sprague-Dawley rats. Endocrinology 136:2320–2324

    Article  PubMed  CAS  Google Scholar 

  • Smith DJ, Zhu Y, Zhang J et al (1995) Construction of a panel of transgenic mice containing a contiguous 2-Mb set of YAC/P1 clones from human chromosome 21q22.2. Genomics 27:425–434

    Article  PubMed  CAS  Google Scholar 

  • Snider WD (1994) Functions of the neurotrophins during nervous system development: what the knockouts are teaching us. Cell 77:627–638

    Article  PubMed  Google Scholar 

  • Stocksley MA, Chakkalakal JV, Bradford A et al (2005) A 1.3 kb promoter fragment confers spatial and temporal expression of utrophin A mRNA in mouse skeletal muscle fibers. Neuromuscul Disord 15:437–449

    Article  PubMed  Google Scholar 

  • Timmusk T, Palm K, Metsis M et al (1993) Multiple promoters direct tissue-specific expression of the rat BDNF gene. Neuron 10:475–489

    Article  PubMed  CAS  Google Scholar 

  • Timmusk T, Lendahl U, Funakoshi H et al (1995) Identification of brain-derived neurotrophic factor promoter regions mediating tissue-specific, axotomy-, and neuronal activity-induced expression in transgenic mice. J Cell Biol 128:185–199

    Article  PubMed  CAS  Google Scholar 

  • Uchida K (1992) Recombination and amplification of multiple portions of genomic DNA by a modified polymerase chain reaction. Anal Biochem 202:159–161

    Article  PubMed  CAS  Google Scholar 

  • Vaidya VA, Marek GJ, Aghajanian GK et al (1997) 5-HT2A receptor-mediated regulation of brain-derived neurotrophic factor mRNA in the hippocampus and the neocortex. J Neurosci 17:2785–2795

    PubMed  CAS  Google Scholar 

  • Wu D, Pardridge WM (1999) Neuroprotection with noninvasive neurotrophin delivery to the brain. Proc Natl Acad Sci USA 96:254–259

    Article  PubMed  CAS  Google Scholar 

  • Yan Q, Rosenfeld RD, Matheson CR et al (1997) Expression of brain-derived neurotrophic factor protein in the adult rat central nervous system. Neuroscience 78:431–448

    Article  PubMed  CAS  Google Scholar 

  • Yuen EC, Mobley WC (1996) Therapeutic potential of neurotrophic factors for neurological disorders. Ann Neurol 40:346–354

    Article  PubMed  CAS  Google Scholar 

  • Zhou AW, Li WX, Guo J et al (1997) Facilitation of AVP(4–8) on gene expression of BDNF and NGF in rat brain. Peptides 18:1179–1187

    Article  PubMed  CAS  Google Scholar 

  • Zhuo L, Sun B, Zhang CL et al (1997) Live astrocytes visualized by green fluorescent protein in transgenic mice. Dev Biol 187:36–42

    Article  PubMed  CAS  Google Scholar 

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Acknowledgements

We wish to thank Esther Toselli and Cécile Hervieux for DNA sequencing, Arlette Loeuillet, René Papion-Duchateau, Jean-Paul Moussu and Patrice Medina from SEAT for technical assistance in trangenesis and mouse manipulation, Nathalie Spassky and Bernard Zalc (INSERM U-495, Hopital de la Salpêtrière, Paris) for help in E-GFP visualization, MAP2 detection and interest for this work. We are indebtful to Clare Huxley for advices and sharing protocols. This work was supported by grants from Association Française contre les Myopathies (AFM) and Ligue Nationale contre le Cancer. The authors are grateful to Association France-Alzheimer for interest and financial support provided to FG. FG also benefited from fellowships from AFM, Ligue Nationale contre le Cancer, Association pour la Recherche contre le Cancer and from Cold Spring Harbor Laboratory concerning the “YACs in Structural & Biological Genome Analysis” course.

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Authors and Affiliations

  1. Genexpress, Génomique Fonctionnelle et Biologie Systémique pour la Santé, CNRS et Université Pierre et Marie Curie Paris VI, LGN, UMR 7091, 7 rue Guy Moquet, BP8, 94801, Villejuif, France

    Fabrice Guillemot, Charles Auffray & Marie-Dominique Devignes

  2. Service d’Expérimentation Animale et de Transgenèse, SEAT, CNRS UPS 44, 94801, Villejuif, France

    Italina Cerutti

  3. Laboratoire Lorrain en Informatique et ses Applications, LORIA, CNRS UMR 7503, 54500, Vandoeuvre-les Nancy, France

    Marie-Dominique Devignes

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  1. Fabrice Guillemot
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  2. Italina Cerutti
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  3. Charles Auffray
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  4. Marie-Dominique Devignes
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Correspondence to Charles Auffray.

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Guillemot, F., Cerutti, I., Auffray, C. et al. A transgenic mouse model engineered to investigate human brain-derived neurotrophic factor in vivo. Transgenic Res 16, 223–237 (2007). https://doi.org/10.1007/s11248-006-9060-0

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  • DOI: https://doi.org/10.1007/s11248-006-9060-0

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