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Topographical localisation of endothelin mRNA and peptide immunoreactivity in neurones of the human brain

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The distribution of endothelin mRNA and immunoreactivity in the human brain was investigated using the technique of in situ hybridization and immunocytochemistry. Cryostat sections from 22 cases of neurologically normal adult human brain, collected 3–7 h post-mortem were hybridized with35S-labelled complementary (c)RNA probes prepared from the 3′ non-coding region of endothelin-1 cDNA, and the chromosomal genes encoding endothelin-2 and -3. In situ hybridization with all three cRNA probes revealed labelled neuronal cell bodies in laminae III–VI of the parietal, temporal and frontal cortices. Labelled cells were also seen, scattered throughout the para- and periventricular; supraoptic and lateral hypothalamic nuclei, the caudate nucleus, amygdala, hippocampus, basal nucleus of Meynert, substantia nigra, raphe nuclei, Purkinje cell layer of the cerebellum and in the dorsal motor nuclei of the vagus of the medulla oblongata. The distribution of neurones immunoreactive to endothelin was similar to that of endothelin mRNA, although fewer immunoreactive cells throughout the brain, were noted. Immunoreactive fibres were present mainly in the cortex and hypothalamus, and to a lesser extent in the brain stem. Combined in situ hybridization and immunocytochemistry on the same section revealed the presence of endothelin-1 mRNA and immunoreactivity in the same cortical neuronal cell. Colocalisation studies in the cortex revealed endothelin-1 mRNA and immunoreactivity in a number of cells which also expressed neuropeptide Y mRNA and immunoreactivity. In the hypothalamus and basal nucleus of Meynert endothelin immunoreactivity was colocalised to a subset of neurophysin- and galanin-immunoreactive cell bodies respectively. Endothelin mRNA and immunoreactivity was also seen in some blood vessel endothelial cells. The findings of endothelin mRNAs and immunoreactivity in heterogenous neuronal populations further emphasises the potential role of endothelin as a neuropeptide, probably having diverse actions in the nervous system of man.

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References

  • Allen YS, Adrian TE, Allen JM, Tatemoto K, Crow TJ, Bloom SR, Polak JM (1983) Neuropeptide Y distribution in the rat brain. Science 221:877–879

    Google Scholar 

  • Ambar I, Kloog Y, Schavartz I, Hazum E, Sokolovsky N (1989) Competitive interaction between endothelin and sarafotoxin binding and phosphoinositide hydrolysis in rat atria and brain. Biochem Biophys Res Commun 158:195–201

    Google Scholar 

  • Beach TG, Tago H, Nagai T, Kimura H, McGeer PL, McGeer EG (1987) Perfusion-fixation of the human brain for immunohistochemistry: comparison with immersion-fixation. J Neurosci Methods 19:183–192

    Google Scholar 

  • Bredt DS, Hwang PM, Snyder SH (1990) Neural and endothelial localizations of nitric oxide synthase in the rat. Nature. (in press)

  • Brene S, Lindefors N, Kopp J, Sedvall G, Persson H (1989) Regional distribution of neuropeptide Y mRNA in post-mortem human brain. Mol Brain Res 6:241–249

    Google Scholar 

  • Bobik A, Grooms A, Millar JA, Mitchell A, Grinpukel S (1990) Growth factor activity of endothelin on vascular smooth muscle. Am J Physiol 259:408–415

    Google Scholar 

  • Calvo JJ, Gonzalez R, Carvalho LF, Takahashi K, Kanse SM, Hart GR, Ghatei MA, Bloom SR (1990) Release of substance P from rat hypothalamus and pituitary by endothelin. Endocrinology 126:2288–2295

    Google Scholar 

  • Chan-Palay V, Allen YS, Lang W, Haesler U, Polak JM (1985a) 1. Cytology and distribution in normal human cerebral cortex of neurons immunoreactive with antisera against neuropeptide Y. J Comp Neurol 238:382–389

    Google Scholar 

  • Chan-Palay V, Lang W, Allen YS, Haesler U, Polak JM (1985b) Cortical neurons immunoreactive with antisera against neuropeptide Y and its precursor peptide are altered in Alzheimer's dementia. J Comp Neurol 238:390–400

    Google Scholar 

  • Chan-Palay V (1988) Galanin hyperinnervates surviving neurons of the human basal nucleus of Meynert in dementias of Alzheimer's and Parkinson's disease: a hypothesis for the role of galanin in accentuating cholinergic dysfunction in dementia. J Comp Neurol 273:543–557

    Google Scholar 

  • Cintra A, Fuxe K, Anggard E, Tinner B, Staines W, Agnati F (1989) Increased endothelin-like immunoreactivity in ibotenic acid-lesioned hippocampal formation of the rat brain. Acta Physiol Scand 137:557–558

    Google Scholar 

  • Escrig C, Bishop AE, Inagaki H, Moscoso G, Polak JM (1990) Endothelin-1: endothelin-like immunolocalization in adult and developing gut. Pathol Soc Great Britain and Ireland, July 10–13th, p. 159

  • Fuxe K, Cintra A, Andbjer B, Anggard E, Goldstein M, Agnati LF (1989) Centrally administrated endothelin-1 produces lesions in the brain of the male rat. Acta Physiol Scand 137:155–156

    Google Scholar 

  • Gentleman SM, Falkai P, Bogerts B, Herrero MT, Polak JM, Roberts GW (1989) Distribution of galanin-like immunoreactivity in the human brain. Brain Res 505:311–315

    Google Scholar 

  • Giaid A, Gibson SJ, IBrahim NBN, Legon S, Bloom SR, Yanagisawa M, Masaki T, Varndell IM, Polak JM (1989) Endothelin 1, an endothelium-derived peptide, is expressed in neurones of the human spinal cord and dorsal root ganglia. Proc Natl Acad Sci USA 86:7634–7638

    Google Scholar 

  • Giaid A, Polak JM, Gaitonde V, Hamid QA, Moscoso G, Legon S, Uwanogho D, Roncalli M, Shinmi O, Sawamura T, Kimura S, Yanagisawa M, Masaki T, Springall DR (1990) The distribution of endothelin-like immunoreactivity and mRNA in the developing and adult human lung. Am Respir Dis (in press)

  • Gibson SJ, Polak JM, Bloom SR, Wall PD (1981) The distribution of nine peptides in rat spinal cord with special emphasis on the substantia gelatinosa and on the area around the central canal (Lamina X). J Comp Neurol 201:65–79

    Google Scholar 

  • Gibson SJ, Polak JM, Bloom SR, Sabate IM, Mulderry PM, Ghatei MA, Morrison JFB, Kelly JS, Evans R, Rosenfeld MG (1984) Calcitonin gene-related peptide (CGRP) immunoreactivity in the spinal cord of man and of eight other species. J Neurosci 4:3101–3111

    Google Scholar 

  • Gibson SJ, Polak JM (1986) Neurochemistry of the spinal cord. In: Polak JM, Van Noorden S. Immunocytochemistry. Modern methods and applications, 2nd edn. John Wright & Sons, Bristol, UK, pp 360–389

    Google Scholar 

  • Gibson SJ, Polak JM, Giaid A, Hamid QA, Kar S, Jones PM, Denny P, Legon S, Amara SG, Craig RK, Bloom SR, Penketh RJA, Rodek C, Ibrahim NBN, Dawson A (1988) Calcitonin gene-related peptide messenger RNA is expressed in sensory neurones of the dorsal root ganglia and also in spinal motoneurones in man and rat. Neurosci Lett 91:283–288

    Google Scholar 

  • Hamid Q, Wharton J, Terenghi G, Hassall CTS, Aimi J, Taylor KM, Nakazato H, Dixon JE, Burnstock G, Polak JM (1987) Localisation of atrial natriuretic peptide mRNA and immunoreactivity in the rat heart and human atrial appendage. Proc Natl Acad Sci USA 84:6760–6764

    Google Scholar 

  • Hirata Y, Matsungaga T, Ando K, Furukawa T, Tsukagashi H, Marumo F (1990) Presence of endothelin-1-like immunoreactivity in human cerebropsinal fluid. Biochem Biophys Res Commun 166:1274–1278

    Google Scholar 

  • Hoefler H, Childers H, Montminy MR, Lechan RM, Goodman RH, Wolfe HJ (1986) In situ hybridization methods for the detection of somatostatin mRNA in tissue sections using antisense RNA probes. Histochem J 18:596–604

    Google Scholar 

  • Hoyer D, Waeber D, Palacios JM (1989) [125I] Endothelin-1 binding sites: autoradiographic studies in the brain and periphery of various species including humans. J Cardiovasc Pharmacol 13 [Suppl]:S162-S165

    Google Scholar 

  • Hsu SM, Raine L, Fanger H (1981) The use of avidin-biotin peroxidase complex (ABC) in immunoperoxidase technique: a comparison between ABC and unlabelled antibody (PAP) procedures. J Histochem Cytochem 29:577–580

    Google Scholar 

  • Huang WM, Gibson SJ, Facer P, Gu J, Polak JM (1983) Improved section adhesion for immunocytochemistry using high molecular weight polymers ofl-lysine as a slide coating. Histochemistry 77:275–279

    Google Scholar 

  • Inoue A, Yanagisawa M, Kimura S, Kasuya Y, Miyauchi T, Goto K, Masaki T (1989) The human endothelin family: three structurally and pharmacologically distinct isopeptides predicted by three separate genes. Proc Natl Acad Sci USA 86:2863–2867

    Google Scholar 

  • Jones CR, Hiley CR, Pelton JT, Mohr M (1989) Autoradiographic visualization of the binding sites for [125I] endothelin in rat and human brain. Neurosci Lett 97:276–279

    Google Scholar 

  • Komuro I, Kurihara H, Sugiyama T, Takaku T, Yazaki Y (1988) Endothelin stimulates c-fos and c-myc expression and proliferation of vascular smooth muscle cells. FEBS Lett 238:249–252

    Google Scholar 

  • Koseki C, Imai M, Hirata Y, Yanagisawa M, Masaki T (1989) Autoradiographic distribution in rat tissues of binding sites for endothelin: a neuropeptide. Am Physiol Soc 256:858–866

    Google Scholar 

  • Lin WW, Lee CY, Chuang D-M (1989) “Cross-densensitization” of endothelin- and sarafotoxin-induced phosphoinositide turnover in neurons. Eur J Pharmacol 166:581–582

    Google Scholar 

  • MacCumber MW, Ross CA, Snyder SH (1989) Endothelin-visualizatoin of messenger RNA by in situ hybridization provides evidence for local action. Proc Natl Acad Sci USA 86:2785–2789

    Google Scholar 

  • MacCumber MW, Ross CA, Snyder H (1990) Endothelin in brain: receptors mitogenesis, and biosynthesis in glial cells. Proc Natl Acad Sci USA 87:2359–2363

    Google Scholar 

  • Marti E, Gibson SJ, Polak JM, Facer P, Springall DR, Van Aswegen G, Aitchison M, Koltzenburgh M (1987) Ontogeny of peptide- and amine-containing neurones in motor, sensory and autonomic regions of rat and human spinal cord, dorsal root ganglia and rat skin. J Comp Neurol 226:332–359

    Google Scholar 

  • Matsumoto H, Suzuki N, Onda H, Fujino M (1989) Abundance of endothelin-3 in rat intestine, pituitary gland and brain. Biochem Biophys Res Commun 164:74–80

    Google Scholar 

  • Moncada S, Radomski MW, Palmer RMJ (1988) Endotheliumderived relaxing factor. Identification as nitric oxide and role in the control of vascular tone and platelet function. Biochem Pharmacol 37:2495–2501

    Google Scholar 

  • Moser PC, Pelton JT (1990) Behavioural effects of centrally administered endothelin in the rat. Br J Pharmacol (in press)

  • Nieuwenhuys R, Voogd J, Huijzen Chr van (1988) The human nervous system. A synopsis and atlas, 3rd revised edn. Springer, Berlin Heidelberg New York, pp 65–141

    Google Scholar 

  • Palkovits M (1984) Distribution of neuropeptides in the central nervous system: a review of biochemical mapping studies. Prog Neurobiol 23:151–189

    Google Scholar 

  • Power RF, Wharton J, Zhao Y, Bloom SR, Polak JM (1989) Autoradiographic localization of endothelin-1 binding sites in the cardiovascular and respiratory system. J Cardiovasc Pharmacol 13:550–556

    Google Scholar 

  • Rozengurt N, Springall DR, Polak JM (1990) Endothelin-like immunoreactivity is localised in airway epithelium of rat and mouse lung. J Pathol 160:5–8

    Google Scholar 

  • Shichiri M, Hirata Y, Kanno K, Ohta K, Emori T, Marumo F (1989) Effect of endothelin-1 on release of arginine-vasopressin from perfused rat hypothalamus. Biochem Biophys Res Commun 163:1332–1337

    Google Scholar 

  • Shinmi O, Kimura S, Yoshizawa T, Sawamura T, Uchiyama Y, Sugita Y, Kanazawa I, Yanagisawa M, Goto T, Masaki T (1989) Presence of endothelin-1 in porcine spinal cord: isolation and sequence determination. Biochem Biophys Res Commun 162:340–346

    Google Scholar 

  • Shu S, Ju G, Fan L (1988) The glucose oxidase-DAB-nickel method in peroxidase histochemistry of the nervous system. Neurosci Lett 85:169–171

    Google Scholar 

  • Simonson MS, Wann S, Mene P, Dubyak GR, Kester M, Nakazoto Y, Sedor JR, Dunn MJ (1989) Endothelin stimulates pospholipase C, Na+/H+ exchange, c-fos expression and mitogenesis in rat mesangial cells. J Clin Invest 83:708–712

    Google Scholar 

  • Stephenson TJ, Griffiths DWR, Mills PM (1986) Comparison ofUlex europaeus I lectin binding and factor VIII-related antigen as markers of vascular endothelium in follicular carcinoma of the thyroid. Histopathology 10:251–260

    Google Scholar 

  • Snyder S (1980) Brain peptides as neurotransmitters. Science 209:976–983

    Google Scholar 

  • Takahashi K, Ghatei M, Jones PM, Murphy JK, Lam H-C, O'Halloran DJ, Bloom SR (1990) Endothelin in human brain and pituitary gland: presence of immunoreactive endothelin, endothelin mRNA and endothelin receptors. J Clin Endocrinol Metab (in press)

  • Takuwa N, Takuwa Y, Yanagisawa M, Yamashita K, Masaki T (1989) A novel vasoactive peptide endothelin stimulates mitogenesis through inositol lipid turnover in Swiss 3T3 fibroblasts. J Biol Chem 264:7856–7861

    Google Scholar 

  • Terenghi G, Polak JM, Hamid Q, O'Brian E, Denny P, Legon S, Dixon J, Minth CD, Palay SL, Yasargil G, Chan-Palay V (1987) Localization of neuropeptide Y mRNA in neurones of human cerebral cortex by means of in situ hybridisation with a complementary RNA probe. Proc Natl Acad Sci USA 84:7315–7318

    Google Scholar 

  • Watkins WB (1976) Immunohistochemical study of the hypothalamo-neurohypophyseal system. I. Localisation of neurosecretory neurons containing neurophysin-I and neurophysin-II in domestic pig. Cell Tissue Res 175:165–181

    Google Scholar 

  • Yanagisawa M, Kurihara H, Kimura S, Tomobe Y, Kobayashi M, Mitsui Y, Yazaki Y, Goto K, Masaki T (1988a) A novel potent vasoconstrictor peptide produced by vascular endothelial cells. Nature 332:411–415

    Google Scholar 

  • Yanagisawa M, Inoue A, Ishikawa T, Kasuya Y, Kimura S, Kumagaye S-I, Nakajima K, Watanabe TX, Sakakibara S, Goto K, Masaki T (1988b) Primary structure, synthesis, and biological activity of rat endothelin, an endothelium-derived vasoconstrictor peptide. Proc Natl Acad Sci USA 85:6964–6967

    Google Scholar 

  • Yanagisawa M, Masaki T (1989) Molecular biology and biochemistry of the endothelins. Trends Neurosci 101:374–378

    Google Scholar 

  • Yoshimoto S, Ishizaki Y, Kurihara H, Sasaki T, Yoshizumi M, Yanagisawa M, Yazaki Y, Masaki T, Takura M, Murota S-I (1990) Cerebral microvessel endothelium is producing endothelin. Brain Res 508:283–285

    Google Scholar 

  • Yoshizawa I, Kimura S, Kamazawa I, Uchiyama Y, Yanagisawa M, Masaki T (1989) Endothelin localizes in the dorsal horn and acts on the spinal neurones: possible involvement of dihydropyridine-sensitive calcium channels and substance P release. Neurosci Lett 102:179–184

    Google Scholar 

  • Yoshizawa T, Shinmi O, Giaid A, Yanagisawa M, Gibson SJ, Kimura S, Uchiyama Y, Polak JM, Masaki T, Kanazawa I (1990) Endothelin: a novel peptide in the posterior pituitary system. Science 247:462–463

    Google Scholar 

  • Zhang W, Sakai N, Yamada H, Fu T, Nozawa Y (1990) Endothelin-1 induces intracellular calcium rise and mositol 1,4,5-triphosphate formation in cultured rat and human glioma cells. Neurosci Lett 112:199–204

    Google Scholar 

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Giaid, A., Gibson, S.J., Herrero, M.T. et al. Topographical localisation of endothelin mRNA and peptide immunoreactivity in neurones of the human brain. Histochemistry 95, 303–314 (1991). https://doi.org/10.1007/BF00745003

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