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Both upstream and intragenic sequences of the human neurofilament light gene direct expression oflacZ in neurons of transgenic mouse embryos

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Abstract

Initial expression of the neurofilament light gene coincides with the appearance of postmitotic neurons. To investigate the molecular mechanisms involved in neuron-specific gene expression during embryogenesis, we generated transgenic mice carrying various regions of the human neurofilament light gene (hNF-L) fused to thelacZ reporter gene. We found that 2.3 or 0.3 kb of the hNF-L promoter region directs expression oflacZ in neurons of transgenic embryos. Addition of 1.8 kb hNF-L intragenic sequences (IS) enlarges the neuronal pattern of transgene expression. The 2.3-kb hNF-L promotelacZ-IS construct contains all regulatory elements essential for both spatial and temporal expression of the hNF-L gene during embryogenesis and in the adult. The use of a heterologous promoter demonstrated that the 1.8-kb hNF-L intragenic sequences are sufficient to direct the expression oflacZ in a NF-L-specific manner both temporally and spatially during development and in the adult. We conclude that these hNF-L intragenic sequences containcis-acting DNA regulatory elements that specify neuronal expression. Taken together, these results show that the neurofilament light gene contains separate upstream and intragenic elements, each of which directslacZ expression in embryonic neurons.

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References

  • Altshuler D. M., Turner D. L., and Cepko C. L. (1991) Specification of cell type in the vertebrate retina, inDevelopment of the Visual System (Lam D. M. K. and Shatz C. J., eds.), MIT Press, Cambridge, MA, pp. 37–58.

    Google Scholar 

  • Aronow B., Lattier D., Silbiger R., Dusing M., Hutton J., Jones G., Stock J., McNeish J., Potter S., Witte D., and Wiginton D. (1989) Evidence for a complex regulatory array in the first intron of the human adenosine deaminase gene.Genes Dev. 3, 1384–1400.

    Article  PubMed  CAS  Google Scholar 

  • Beaudet L., Charron G., Houle D., Tretjakoff I., Peterson A., and Julien J. P. (1992) Intragenic regulatory elements contribute to transcriptional control of the neurofilament light gene.Gene 116, 205–214.

    Article  PubMed  CAS  Google Scholar 

  • Begemann M., Tan S. S., Cunningham B. A., and Edelman G. M. (1990) Expression of chicken liver cell adhesion molecule fusion genes in transgenic mice.Proc. Natl. Acad. Sci. USA 87, 9042–9046.

    Article  PubMed  CAS  Google Scholar 

  • Bejanin S., Cervini R., Mallet J., and Berrard S. (1994) A unique gene organization for two cholinergic markers, choline acetyltransferase and a putative vesicular transporter of acetylcholine.J. Biol. Chem. 269 (35), 21,944–21,947.

    CAS  Google Scholar 

  • Belecky-Adams T., Wight D. C., Kopchick J. J. and Parysek L. M. (1993) Intragenic sequences are required for cell type-specific and injury-induced expression of the rat peripherin gene.J. Neurosci. 13 (12), 5056–5065.

    PubMed  CAS  Google Scholar 

  • Bellen H. J., Wilson C., and Gehring W. J. (1990) Dissecting the complexity of the nervous system by enhancer detection.Bioassays 10, 199–204.

    Article  Google Scholar 

  • Bennett G. S. and DiLullo C. (1985a) Expression of a neurofilament protein by precursors of a subpopulation of ventral spinal cord neurons.Dev. Biol. 107, 94–106.

    Article  PubMed  CAS  Google Scholar 

  • Bennett G. S. and DiLullo C. (1985b) Transient expression of a neurofilament protein by replicating neuroepithelial cells of the embryonic chick brain.Dev. Biol. 107, 107–127.

    Article  PubMed  CAS  Google Scholar 

  • Berthoud H. R. and Powley T. L. (1992) Vagal afferent innervation of the rat fundic stomach: morphological characterization of the gastric tension receptor.J. Comp. Neurol. 319, 261–276.

    Article  PubMed  CAS  Google Scholar 

  • Bier E., Vaessin H., Sheperd S., Lee K., McCall K., and Barbel S. (1989) Searching for pattern and mutation in theDrosophila genome with a P-lacZ vector.Genes Dev. 3, 1273–1287.

    Article  PubMed  CAS  Google Scholar 

  • Bonnerot C., Grimber G., Briand P., and Nicolas J. F. (1990) Patterns of expression of position-dependent integrated transgenes in mouse embryo.Proc. Natl. Acad. Sci. USA 87, 6331–6335.

    Article  PubMed  CAS  Google Scholar 

  • Bornstein P. and McKay J. (1988) The first intron of the α1(I) collagen gene contains several transcriptional regulatory elements.J. Biol. Chem. 263, 1603–1606.

    PubMed  CAS  Google Scholar 

  • Brinster R. L., Allen J. M., Behringer R. B., Gelinas R. E., and Palmiter R. D. (1988) Introns increase transcriptional efficiency in transgenic mice.Proc. Natl. Acad. Sci. USA 85, 836–840.

    Article  PubMed  CAS  Google Scholar 

  • Carden M. J., Trojanowski J. Q., Schlaepfer W. W., and Lee V. M. Y. (1987) Two-stage expression of neurofilament polypeptides during rat neurogenesis with early establishment of adult phosphorylation patterns.J. Neurosci. 7 (11), 3489–3504.

    PubMed  CAS  Google Scholar 

  • Cochard P. and Paulin D. (1984) Initial expression of neurofilaments and vimentin in the central and peripheral nervous system of the mouse embryoin vivo.J. Neurosci. 4, 2080–2094.

    PubMed  CAS  Google Scholar 

  • Cohen-Tannoudji M., Morello D., and Babinet C. (1992) Unexpected position-dependent expression of H-2 and β2-microglobulin/lacZ transgenes.Mol. Reprod. Dev. 33, 149–159.

    Article  PubMed  CAS  Google Scholar 

  • Cohen-Tannoudji M., Babinet C., and Wassef M. (1994) Early determination of a mouse somatosensory cortex marker.Nature 368, 460–463.

    Article  PubMed  CAS  Google Scholar 

  • Cumberledge S. and Krasnow A. (1993) Intercellular signalling inDrosophila segment formation reconstructed in vitro.Nature 363, 549–551.

    Article  PubMed  CAS  Google Scholar 

  • Dräger U. C., Edwards D. L., and Barnstable C. J. (1984) Antibodies against filamentous components in discrete cell types of the mouse retina.J. Neurosci. 4 (8), 2025–2042.

    PubMed  Google Scholar 

  • Echelard Y., Vassileva G., and McMahon A. P. (1994) Cis-acting regulatory sequences governingWnt-1 expression in the developing mouse CNS.Development 120, 2213–2224.

    PubMed  CAS  Google Scholar 

  • Erickson J. D., Varoqui H., Schafer M. K. H., Modi W., Diebler M. F., Weihe E., Rand J., Eiden L. E., Bonner T. I., and Usdin T. B. (1994) Functional identification of a vesicular acetylcholine transporter and its expression from a “cholinergic” gene locus.J. Biol. Chem. 269 (35), 21,929–21,932.

    CAS  Google Scholar 

  • Escurat M., Djabali K., Grumpel M., Gros F., and Portier M. M. (1990) Differential expression of two neuronal intermediate filament proteins, peripherin and low molecular mass neurofilament protein (NF-L) during the development of the rat.J. Neurosci. 10, 764–784.

    PubMed  CAS  Google Scholar 

  • Evans T., Felsenfeld G., and Reitman M. (1990) Control of globin gene transcription.Annu. Rev. Cell. Biol. 6, 95–124.

    Article  PubMed  CAS  Google Scholar 

  • Forss-Petter S., Danielson P. E., Catsicas S., Battenberg E., Price J., Nerenberg M., and Sutcliffe J. G. (1990) Transgenic mice expressing β-galactosidase in mature neurons under neuron-specific enolase promoter control.Neuron 5, 187–197.

    Article  PubMed  CAS  Google Scholar 

  • Furness J. B. and Costa M. (1980) Commentary. Types of nerves in the enteric nervous system.Neuroscience 5, 1–20.

    Article  PubMed  CAS  Google Scholar 

  • Godement P., Vanselow J., Thanos S., and Bonhoeffer F. (1987) A study in developing visual systems with a new method of staining neurones and their processes in fixed tissues.Development 101, 697–713.

    PubMed  CAS  Google Scholar 

  • Godement P., Salaun J., and Mason C. (1990) Retinal axon pathfinding in the optic chiasma: divergence of crossed and uncrossed fibers.Neuron 5, 173–186.

    Article  PubMed  CAS  Google Scholar 

  • Hogan B. L. M., Costantini F., and Lacy E. (1986)Manipulating the Mouse Embryo: A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.

    Google Scholar 

  • Hoyle G. W., Mercer E. H., Palmiter R. D., and Brinster R. L. (1994) Cell-specific expression from the human dopamine β-hydroxylase promoter in transgenic mice is controlled via a combination of positive and negative regulatory elements.J. Neurosci. 14, 2455–2463.

    PubMed  CAS  Google Scholar 

  • Jacobson M. (1991)Developmental Neurobiology. 3rd ed. The University of Utah School of Medicine, Salt Lake City, UT.

    Google Scholar 

  • Julien J. P., Grosveld F., Yazdanbakhsh K., Flavell D., Meijer D., and Mushynski W. (1987a) The structure of a human neurofilament gene (NF-L): a unique exon-intron organisation in the intermediate filament gene family.Biochem. Biophys. Acta 909, 10–20.

    PubMed  CAS  Google Scholar 

  • Julien J. P., Tretjakoff I., Beaudet L., and Peterson A. (1987b) Expression and assembly of a human neurofilament protein in transgenic mice provide a novel neuronal marking system.Genes Dev. 1, 1085–1095.

    Article  PubMed  CAS  Google Scholar 

  • Julien J. P., Beaudet L., Tretjakoff I., and Peterson A. (1990) Neurofilament gene expression in transgenic mice.J. Physiol. Paris 84, 50–52.

    PubMed  CAS  Google Scholar 

  • Kapur R. P., Hoyle G. W., Mercer E. H., Brinster R. L., and Palmiter R. D. (1991) Some neuronal cell populations express human dopamine β-hydroxylase-lacZ transgenes transiently during embryonic development.Neuron 7, 717–727.

    Article  PubMed  CAS  Google Scholar 

  • Kapur R. P., Yost C., and Palmiter R. D. (1992) A transgenic model for studying development of the enteric nervous system in normal and aganglionic mice.Development 116, 167–175.

    PubMed  CAS  Google Scholar 

  • Kaufman M. H. (1992)The Atlas of Mouse Development, Academic, San Diego.

    Google Scholar 

  • Kessel M. (1993) Reversal of axonal pathways from rhombomere 3 correlates with extraHox expression domains.Neuron 10, 379–393.

    Article  PubMed  CAS  Google Scholar 

  • Kothary R., Clapoff S., Darling S., Perry M. D., Moran L. A., and Rossant J. (1989) Inducible expression of an hsp68-lacZ hybrid gene in transgenic mice.Development 105, 707–714.

    PubMed  CAS  Google Scholar 

  • Krasnow M. A., Cumberledge S., Manning G., Herzenberg L., and Nolan G. P. (1991) Whole animal cell sorting ofDrosophila embryos.Science 251, 81–85.

    Article  PubMed  CAS  Google Scholar 

  • Logan C., Khoo W. K., Cado D., and Joyner A. L. (1993) Two enhancer regions in the mouseEn-2 locus direct expression to the mid/hindbrain region and mandibular myoblasts.Development 117, 905–916.

    PubMed  CAS  Google Scholar 

  • Nakahira K., Ikenaka K., Wada K., Tamura T., Furuichi T., and Mikoshiba K. (1990) Structure of the 68-kDa neurofilament gene and regulation of its expression.J. Biol. Chem. 265, 19,786–19,791.

    CAS  Google Scholar 

  • Nolan G. P., Fiering S., Nicolas J. F., and Herzenberg L. A. (1988) Fluorescence-activated cell analysis and sorting of viable mammalian cells based on β-d-galactosidase activity after transduction ofEscherichia coli lacZ.Proc. Natl. Acad. Sci. USA 84, 9123–9127.

    Google Scholar 

  • Oshima R. G., Abrams L., and Kulesh D. (1990) Activation of an intron enhancer within the keratin 18 gene by expression ofc-fos andc-jun in undifferentiated F9 embryonal carcinoma cells.Genes Dev. 4, 835–848.

    Article  PubMed  CAS  Google Scholar 

  • Palmiter R. D., Sandgren E. P., Avarbock M. R., Allen D. D., and Brinster R. L. (1991) Heterologous introns can enhance expression of transgenes in mice.Proc. Natl. Acad. Sci. USA 88, 478–482.

    Article  PubMed  CAS  Google Scholar 

  • Pospelov V. A., Pospelova T. V., and Julien J. P. (1994) AP-1 and Krox-24 transcription factors activate the neurofilament light gene promoter in P19 embryonal carcinoma cells.Cell Growth Differ. 5, 187–196.

    PubMed  CAS  Google Scholar 

  • Püschel A. W., Balling R., and Gruss P. (1991) Separate elements cause lineage restriction and specific boundaries ofHox-1.1 expression.Development 112, 279–287.

    PubMed  Google Scholar 

  • Reitman M., Lee E., Westphal H., and Felsenfeld G. (1990) Site-independent expression of the chicken βa-globin gene in transgenic mice.Nature 348, 749–752.

    Article  PubMed  CAS  Google Scholar 

  • Rossant J., Zirngibl R., Cado D., Shago M., and Giguere V. (1991) Expression of a retinoic acid response element-hsp-lacZ transgene defines specific domains of transcriptional activity during mouse embryogenesis.Genes Dev. 5, 1333–1344.

    Article  PubMed  CAS  Google Scholar 

  • Rugh R. (1990)The Mouse: Its Reproduction and Development. Oxford University Press, NY.

    Google Scholar 

  • Sanes J. R., Rubenstein L. R., and Nicolas J. F. (1986) Use of a recombinant retrovirus to study postimplantation cell lineage in mouse embryos.EMBO J. 5, 3133–3142.

    PubMed  CAS  Google Scholar 

  • Schambra U. B., Lauder J. M., and Silver J. (1992)Atlas of the Prenatal Mouse Brain. Academic, San Diego, CA.

    Google Scholar 

  • Shneidman P. S., Bruce J., Schwartz M. L., and Schlaepfer W. W. (1992) Negative regulatory regions are present upstream in the three mouse neurofilament genes.Mol. Brain Res. 13, 127–138.

    Article  PubMed  CAS  Google Scholar 

  • Tapscott S. J., Bennett G. S., and Holtzer H. (1981) Neuronal precursor cells in the chick neural tube express neurofilament proteins.Nature 292, 836–838.

    Article  PubMed  CAS  Google Scholar 

  • Tuggle C. K., Zakany J., Cianetti L., Peschle C., and Nguyen-Huu C. (1990) Region specific enhancers near two mammalian homeobox genes define adjacent rostrocaudal domains in the central nervous system.Genes Dev. 4, 180–189.

    Article  PubMed  CAS  Google Scholar 

  • Vale R. D., Banker G., and Hall Z. W. (1992) The neuronal cytoskeleton, inMolecular Neurobiology (Hall Z., ed.), Sinauer Associates, Sunderland MA, pp. 247–280.

    Google Scholar 

  • Vandaele S., Nordquist D. T., Feddersen R. M., Tretjakoff I., Peterson A. C., and Orr H. T. (1991)Purkinje cell protein-2 regulatory regions and transgene expression in cerebellar compartments.Genes Dev. 5, 1136–1148.

    Article  PubMed  CAS  Google Scholar 

  • Vogels R., Charite J., De Graaff W., and Deschamps J. (1993) Proximal cis-acting elements cooperate to setHoxb-7 (Hox-2.3) expression boundaries in transgenic mice.Development 118, 71–82.

    PubMed  CAS  Google Scholar 

  • Whiting J., Marshall H., Cook M., Krumlauf R., Rigby P. W. J., Stott D., and Allemann R. K. (1991) Multiple spatially specific enhancers are required to reconstruct the pattern ofHox-2.6 gene expression.Genes Dev. 5, 2048–2059.

    Article  PubMed  CAS  Google Scholar 

  • Wilson C., Bellen H. J., and Gehring W. J. (1990) Position effects on eukaryotic gene expression.Annu. Rev. Cell Biol. 6, 679–714.

    Article  PubMed  CAS  Google Scholar 

  • Yazdanbakhsh K., Fraser P., Kioussis D., Vidal M., Grosveld F., and Lindenbaum M. (1993) Functional analysis of the human neurofilament light chain gene promoter.Nucleic Acids Res. 21, 455–461.

    Article  PubMed  CAS  Google Scholar 

  • Zakany J., Tuggle C. K., Patel M. D., and Nguyen-Huu C. (1988) Spatial regulation of homeobox gene fusions in the embryonic central nervous system of transgenic mice.Neuron 1, 679–691.

    Article  PubMed  CAS  Google Scholar 

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  1. Laurence Leconte

    Present address: Department of Ophthalmology and Visual Science, Yale University School of Medicine, 330 Cedar Street, 06510, New Haven, CT

Authors and Affiliations

  1. Laboratoire de Neurobiologie Cellulaire et Moléculaire, CNRS, Gif-sur-Yvette, France

    Laurence Leconte, Olivier Semonin, Agnès Zvara, Sylvie Boisseau, Chantal Poujeol & Michel Simonneau

  2. Institute of Genetics, Biological Research Center, Hungarian Academy of Sciences, H-6701, Szeged, Hungary

    Agnès Zvara

  3. The Montreal General Hospital Research Institute, H3G 1A4, Montreal, Quebec, Canada

    Jean-Pierre Julien

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  1. Laurence Leconte
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Leconte, L., Semonin, O., Zvara, A. et al. Both upstream and intragenic sequences of the human neurofilament light gene direct expression oflacZ in neurons of transgenic mouse embryos. J Mol Neurosci 5, 273–295 (1994). https://doi.org/10.1007/BF02736727

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