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Expression of human tyrosine hydroxylase-chloramphenicol acetyltransferase (CAT) fusion gene in the brains of transgenic mice as examined by CAT immunocytochemistry

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Summary

We have produced transgenic (Tg) mice carrying 5.0-kb fragment from the 5′-flanking region of the human tyrosine hydroxylase (hTH) gene fused to a reporter gene, chloramphenicol acetyltransferase (CAT) [Sasaoka et al. (1992) Mol Brain Res 16: 274–286]. In the brain of the Tg mice, CAT expression has been observed in catecholaminergic (CAnergic) neurons and also in non-CAnergic neurons. The aim of the present study is to examine in detail the cell-type specific expression of the hTH-CAT fusion gene in the brain of the Tg mice, by use of immunohistochemistry for CAT, TH, and aromatic L-amino acid decarboxylase (AADC). CAT-immunoreactive cells were found in CAnergic brain regions which contained TH-positive cells, and also in non-CAnergic brain regions which contained no TH-labeled cells. The non-CAnergic brain regions that represented CAT-stained cells were further divided into two groups: (i) regions containing AADC-labeled cells, for example, bed nucleus of the stria terminalis, nucleus suprachiasmaticus, mammillary body, nucleus raphe dorsalis, inferior colliculus, and nucleus parabrachialis, and (ii) regions containing no AADC-positive cells, for example, main olfactory bulb (except A16), accessory olfactory bulb, nucleus olfactorius anterior, caudoputamen, septum, nucleus accumbens, hippocampus, medial nucleus of the amygdala, entorhinal cortex, nucleus supraopticus, and parasubiculum. The results indicate that the 5.0-kb DNA fragment flanking the 5′ end of the hTH gene may contain the element(s) specific for neuron-specific TH expression but which may be insufficient to attenuate ectopic expression.

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References

  • Austin MC, Schultzberg M, Abbott LC, Montpied P, Evers JR, Paul S, Crawley JN (1992) Expression of tyrosine hydroxylase in cerebellar Purkinje neurons of the mutant tottering and leaner mouse. Mol Brain Res 15: 227–240

    Google Scholar 

  • Banerjee SA, Hoppe P, Brilliant M, Chikaraishi DM (1992) 5′ Flanking sequences of the rat tyrosine hydroxylase gene target accurate tissue-specific, developmental, and transsynaptic expression in transgenic mice. J Neurosci 12: 4460–4467

    Google Scholar 

  • Berger B, Verney C, Caspar P, Febvret A (1985) Transient expression of tyrosine hydroxylase immunoreactivity in some neurons of the rat neocortex during postnatal development. Dev Brain Res 23: 141–144

    Google Scholar 

  • Coker GTIII, Vinnedge L, O'Malley KL (1988) Characterization of rat and human tyrosine hydroxylase genes: functional expression of both promoters in neuronal and non-neuronal cell types. Biochem Biophys Res Commun 157: 1341–1347

    Google Scholar 

  • Combi F, Fung B, Chikaraishi D (1989) 5′ Flanking DNA sequences direct cell-specific expression of rat tyrosine hydroxylase. J Neurochem 53: 1656–1659

    Google Scholar 

  • Dahlström A, Fuxe K (1964) Evidence for the existence of monoamine-containing neurons in the central nervous system. I. Demonstration of monoamines in the cell bodies of brain stem neurons. Acta Physiol Scand 62 [Suppl] 232: 1–55

    Google Scholar 

  • D'Mello SR, Turzan LM, Gioio AE, Kaplan BB (1989) Isolation and structural characterization of the bovine tyrosine hydroxylase gene. J Neurosci Res 23: 31–40

    Google Scholar 

  • Fujii T, Komori K, Sakai M, Yamada K, Karasawa N, Miura K, Nagatsu I (1992) Immunocytochemical study on transient expression of tyrosine hydroxylase-immunoreactive neurons in the mouse telencephalon during postnatal development. Biogenic Amines 9: 115–122

    Google Scholar 

  • Gizang-Ginsberg E, Ziff EB (1990) Nerve growth factor regulates tyrosine hydroxylase gene transcription through a nucleoprotein complex that contains c-Fos. Genes Dev 4: 477–491

    Google Scholar 

  • Grima B, Lamouroux A, Boni C, Julien J-F, Javoy-Azid F, Mallet J (1987) A single human gene encoding multiple tyrosine hydroxylases with different predicted functional characteristics. Nature 326: 707–711

    Google Scholar 

  • Harrington CA, Lewis EJ, Krzemien D, Chikaraishi D (1987) Identification and cell type specificity of the tyrosine hydroxylase gene promoter. Nucl Acids Res 15: 2363–2384

    Google Scholar 

  • Hess EJ, Wilson MC (1991) Tottering and leaner mutations perturb transient developmental expression of tyrosine hydroxylase in embryologically distinct Purkinje cells. Neuron 6: 123–132

    Google Scholar 

  • Hökfelt T, Martensson R, Björklund A, Kleinau S, Goldstein M (1984) Distributional maps of tyrosine hydroxylase-immunoreactive neurons in the rat brain. In: Björklund A, Hökfelt T (eds) Handbook of chemical neuroanatomy. Classical transmitters in the CNS, vol 2, part 1. Elsevier, Amsterdam, pp 277–379

    Google Scholar 

  • Ichikawa S, Sasaoka T, Nagatsu T (1991) Primary structure of mouse tyrosine hydroxylase deduced from its cDNA. Biochem Biophys Res Commun 176: 1610–1616

    Google Scholar 

  • Ichinose H, Kurosawa Y, Titani K, Fujita K, Nagatsu T (1989) Isolation and characterization of a cDNA clone encoding human aromatic L-amino acid decarboxylase. Biochem Biophys Res Commun 164: 1024–1030

    Google Scholar 

  • Ichinose H, Sumi-Ichinose C, Ohye T, Hagino Y, Fujita K, Nagatsu T (1992) Tissue-specific alternative splicing of the first exon generates two types of mRNAs in human aromatic L-amino acid decarboxylase. Biochemistry 31: 11546–11550

    Google Scholar 

  • Iwata N, Kobayashi K, Sasaoka T, Hidaka H, Nagatsu T (1992) Structure of the mouse tyrosine hydroxylase gene. Biochem Biophys Res Commun 182: 348–354

    Google Scholar 

  • Jaeger CB, Joh TH (1983) Transient expression of tyrosine hydroxylase in some neurons of the developing inferior colliculus of the rat. Dev Brain Res 11: 128–132

    Google Scholar 

  • Jaeger CB, Ruggier DA, Albert VR, Joh TH, Reis DJ (1984) Immunocytochemical localization of aromatic-L-aminoacid decarboxylase. In: Björklund A, Hökfelt T (eds) Handbook of chemical neuroanatomy, vol 2, part 1. Classical transmitters in the CNS. Elsevier, Amsterdam, pp 387–408

    Google Scholar 

  • Kaneda N, Ichinose H, Kobayashi K, Oka K, Kishi F, Nakazawa A, Kurosawa Y, Fujita K, Nagatsu T (1988) Molecular cloning of cDNA and chromosomal assignment of the gene for human phenylethanolamine N-methyltransferase, the enzyme for norepinephrine biosynthesis. J Biol Chem 263: 7672–7677

    Google Scholar 

  • Kaneda N, Kobayashi K, Ichinose H, Kishi F, Nakazawa A, Kurosawa Y, Fujita K, Nagatsu T (1987) Isolation of a novel cDNA clone for human tyrosine hydroxylase: alternative RNA splicing produces four kinds of mRNA from a single gene. Biochem Biophys Res Commun 146: 971–975

    Google Scholar 

  • Kaneda N, Sasaoka T, Kobayashi K, Kiuchi K, Nagatsu I, Kurosawa Y, Fujita K, Yokoyama M, Nomura T, Katsuki M, Nagatsu T (1991) Tissue-specific and high-level expression of the human tyrosine hydroxylase gene in transgenic mice. Neuron 6: 583–594

    Google Scholar 

  • Katoh Y, Shimizu N (1982) The light and electron microscopic localization of intracyto- plasmic nucleolus-like bodies in the mouse brain stained by Holmes' silver method. Arch Histol Jap 45: 325–333

    Google Scholar 

  • Kobayashi K, Kaneda N, Ichinose H, Kishi F, Nakazawa A, Kurosawa Y, Fujita K, Nagatsu T (1987) Isolation of a full length cDNA clone encoding human tyrosine hydroxylase type 3. Nucl Acids Res 75: 6733

    Google Scholar 

  • Kobayashi K, Kaneda N, Ichinose H, Kishi F, Nakazawa A, Kurosawa Y, Fujita K, Nagatsu T (1988 a) Structure of the human tyrosine hydroxylase gene: alternative splicing from a single gene accounts for generation of four mRNA types. J Biochem 103: 907–912

    Google Scholar 

  • Kobayashi K, Kiuchi K, Ishii A, Kaneda N, Kurosawa Y, Fujita K, Nagatsu T (1988 b) Expression of four types of human tyrosine hydroxylase in COS cells. FEBS Lett 238: 431–434

    Google Scholar 

  • Kobayashi K, Kurosawa Y, Fujita K, Nagatsu T (1989) Human dopamine β-hydroxylase gene: two mRNA types having different 3′-terminal regions are produced through alternative polyadenylation. Nucl Acids Res 17: 1089–1102

    Google Scholar 

  • Komori K, Sakai M, Karasawa N, Yamada K, Nagatsu I (1991) Evidence for transient expression of tyrosine hydroxylase immunoreactivity in the mouse striatum and the effect of colchicine. Acta Histochem Cytochem 24: 223–231

    Google Scholar 

  • LeBourdelles B, Boularand S, Boni C, Horellou P, Dumas S, Grima B, Mallet J (1988) Analysis of 5′ region of the human tyrosine hydroxylase gene: combinational patterns of exon splicing generate multiple regulated tyrosine hydroxylase isoforms. J Neurochem 50: 988–991

    Google Scholar 

  • Levitt M, Spector S, Sjoerdsma A, Udenfriend S (1965) Elucidation of the rate-limiting step in norepinephrine biosynthesis in the perfused guinea-pig heart. J Pharmacol Exp Ther 148: 1–8

    Google Scholar 

  • Lewis EJ, Chikaraishi DM (1987) Regulated expression of the tyrosine hydroxylase gene by epidermal growth factor. Mol Cell Biol 7: 3332–3336

    Google Scholar 

  • Lewis EJ, Harrington CA, Chikaraishi DM (1987) Transcriptional regulation of the tyrosine hydroxylase gene by glucocorticoid and cyclic AMP. Proc Natl Acad Sci USA 84: 3550–3554

    Google Scholar 

  • Lewis EJ, Tank AW, Weiner N, Chikaraishi D (1983) Regulation of tyrosine hydroxylase mRNA by glucocorticoid and cyclic AMP in a rat pheochromocytoma cell line. J Biol Chem 258: 14632–14637

    Google Scholar 

  • Morimura S, Suzuki T, Hochi S, Yuki A, Nomura K, Kitagawa T, Nagatsu I, Imagawa M, Muramatsu M (1993) Trans-activation of glutathione transferase P gene during chemical hepatocarcinogenesis of the rat. Proc Natl Acad Sci USA 90: 2065–2068

    Google Scholar 

  • Nagatsu I, Inagaki S, Kondo Y, Karasawa N, Nagatsu T (1979) Immunofluorescent studies on the localization of tyrosine hydroxylase and dopamine-β-hydroxylase in the mes-, di-, and telencephalon of the rat using unperfused fresh frozen sections. Acta Histochem Cytochem 12: 20–37

    Google Scholar 

  • Nagatsu I, Komori K, Takeuchi T, Sakai M, Yamada K, Karasawa N (1990) Transient tyrosine hydroxylase-immunoreactive neurons in the region of the anterior olfactory nucleus of pre- and postnatal mice do not contain dopamine. Brain Res 511: 55–62

    Google Scholar 

  • Nagatsu I, Kondo Y, Inagaki S, Karasawa N, Kato T, Nagatsu T (1977) Immunofluorescent studies on tyrosine hydroxylase: application for its axoplasmic transport. Acta Histochem Cytochem 10: 494–499

    Google Scholar 

  • Nagatsu I, Sakai M, Yoshida M, Nagatsu T (1988) Aromatic L-amino acid decarboxylase-immunoreactive neurons in and around the cerebrospinal fluid-contacting neurons of the central canal do not contain dopamine or serotonin in the mouse and rat spinal cord. Brain Res 475: 91–102

    Google Scholar 

  • Nagatsu I, Yamada K, Karasawa N, Kaneda N, Sasaoka T, Kobayashi K, Fujita K, Nagatsu T (1993) Non-catecholaminergic neuronal expression of human tyrosine hydroxylase in the brain of transgenic mice with special reference to aromatic L-amino acid decarboxylase. In: Naoi M, Palvez HS (eds) Tyrosine hydroxylase. VSP Science Press, Zeist, The Netherlands, pp 37–57

    Google Scholar 

  • Nagatsu I, Yamada K, Karasawa N, Sakai M, Takeuchi T, Kaneda N, Sasaoka T, Kobayashi K, Yokoyama M, Nomura T, Katsuki M, Fujita K, Nagatsu T (1991) Expression in brain sensory neurons of the transgene in transgenic mice carrying human tyrosine hydroxylase gene. Neurosci Lett 127: 91–95

    Google Scholar 

  • Nagatsu T, Levitt M, Udenfriend S (1964) Tyrosine hydroxylase: the initial step in nor-epinephrine biosynthesis. J Biol Chem 239: 2910–2917

    Google Scholar 

  • Nakano T, Kobayashi K, Saito S, Fujita K, Nagatsu T (1992) Mouse dopamine β-hydroxylase: primary structure deduced from the cDNA sequence and exon/intron organization of the gene. Biochem Biophys Res Commun 189: 590–599

    Google Scholar 

  • O'Malley KL, Anhalt MJ, Martin BM, Kelsoe JR, Winfield SL, Ginns EI (1987) Isolation and characterization of the human tyrosine hydroxylase gene: identification of 5′ alternative splice sites responsible for multiple mRNAs. Biochemistry 26: 6910–6914

    Google Scholar 

  • Palmitter RD, Brister RL (1986) Germ-line transformation of mice. Ann Rev Genet 20: 465–499

    Google Scholar 

  • Paxinos G, Watson C (1982) The rat brain in stereotaxis coordinates. Academic Press, New York

    Google Scholar 

  • Rosenfeld MG, Grenshaw EBIII, Lira SA (1988) Transgenic mice: application to the study of the nervous system. Ann Rev Neurosci 11: 353–372

    Google Scholar 

  • Russo AF, Crenshaw EB, Lira SA, Simmon DM, Swanson LW, Rosenfeld MG (1988) Neuronal expression of chimeric genes in transgenic mice. Neuron 1: 311–320

    Google Scholar 

  • Sasaoka T, Kaneda N, Kurosawa Y, Fujita K, Nagatsu T (1989) Structure of human phenylethanolamine N-methyltransferase gene: existence of two types of mRNA with different transcription initiation sites. Neurochem Int 15: 555–565

    Google Scholar 

  • Sasaoka T, Kobayashi K, Nagatsu I, Takahashi R, Kimura M, Yokoyama M, Nomura T, Katsuki M, Nagatsu T (1992) Analysis of the human tyrosine hydroxylase promoterchloramphenicol acetyltransferase chimeric gene expression in trangenic mice. Mol Brain Res 16: 274–286

    Google Scholar 

  • Satoh J, Suzuki K (1990) Tyrosine hydroxylase-immunoreactive neurons in the mouse cerebral cortex during the postnatal period. Dev Brain Res 53: 1–5

    Google Scholar 

  • Sidman RL, Angevine JB, Jr, Pierce ET (1971) Atlas of the mouse brain and spinal cord. Harvard University Press, Massachusetts

    Google Scholar 

  • Sternberger LA (1986) The unlabelled antibody peroxidase-antiperoxidase (PAP) methods. In: Immunocytochemistry, 3rd edn. Wiley, New York, pp 90–209

    Google Scholar 

  • Sumi-Ichinose C, Ichinose H, Takahashi E, Hori T, Nagatsu T (1992) Molecular cloning of genomic DNA and chromosomal assignment of the gene for human aromatic L-amino acid decarboxylase, the enzyme for catecholamine and serotonin biosynthesis. Biochemistry 31: 2229–2238

    Google Scholar 

  • Tank AW, Curella P, Ham L (1986) Induction of mRNA for tyrosine hydroxylase by cyclic AMP and glucocorticoids in a rat pheochromocytoma cell line: evidence for the regulation of tyrosine hydroxylase synthesis by multiple mechanism in cells exposed to elevated levels of both inducing agents. Mol Pharmacol 30: 497–503

    Google Scholar 

  • Verney C, Gasper P, Febvret A, Berger B (1988) Transient tyrosine hydroxylase-like immunoreactive neurons contain somatostatin and substance P in the developing amygdala and bed nucleus of the stria terminalis of the rat. Dev Brain Res 42: 45–58

    Google Scholar 

  • Yamada K, Sakai M, Okamura H, Ibata Y, Nagatsu I (1992) Detection of tyrosine hydroxylase and phenylethanolamine-N-methyltransferase messenger RNAs in the mouse adrenal gland and the brain by in situ hybridization. Histochemistry 97: 201–206

    Google Scholar 

  • Yamada K, Sakai M, Takeuchi T, Karasawa N, Kaneda N, Sasaoka T, Kobayashi K, Yokoyama M, Nomura T, Katsuki M, Fujita K, Nagatsu T, Nagatsu I (1991) Enhanced expression of human tyrosine hydroxylase in the lower brainstem of transgenic mice. Neurosci Lett 134: 57–61

    Google Scholar 

  • Zigmond RE, Schwartzschild MA, Rittenhouse AR (1989) Acute regulation of tyrosine hydroxylase by nerve activity and by neurotransmitters via phosphorylation. Ann Rev Neurosci 12: 415–461

    Google Scholar 

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  1. I. Nagatsu

    Present address: Department of Anatomy, School of Medicine, Fujita Health University, Toyoake, Japan

Authors and Affiliations

  1. Department of Anatomy, School of Medicine, Fujita Health University, Toyoake, Japan

    N. Karasawa, K. Yamada, M. Sakai, T. Fujii, T. Takeuchi & R. Arai

  2. Institute for Comprehensive Medical Science, School of Medicine, Fujita Health University, Toyoake, Japan

    K. Kobayashi & T. Nagatsu

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  1. I. Nagatsu
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  2. N. Karasawa
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  3. K. Yamada
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Nagatsu, I., Karasawa, N., Yamada, K. et al. Expression of human tyrosine hydroxylase-chloramphenicol acetyltransferase (CAT) fusion gene in the brains of transgenic mice as examined by CAT immunocytochemistry. J. Neural Transmission 96, 85–104 (1994). https://doi.org/10.1007/BF01277931

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