Skip to main content
SpringerLink
Log in

Visualisation of specific binding sites for atrial natriuretic peptide on non-neuronal cells of cultured rat sympathetic ganglia

  • Published:
Cell and Tissue Research Aims and scope Submit manuscript

Summary

The distribution of atrial natriuretic peptide binding sites on cells in dissociated culture preparations of neonatal rat superior cervical ganglia and in explant cultures of rat thoracic sympathetic chain ganglia has been studied. The autoradiographic visualisation of atrial natriuretic peptide binding sites has been combined with the use of specific immunocytochemical markers for glial cells (antiserum to S-100 protein), fibroblasts (antiserum to fibronectin) and neurones (antiserum to protein gene product 9.5) in order to achieve unambiguous identification of the cell types in culture. Specific binding sites for rat125I-atrial natriuretic peptide(1–28) were observed over subpopulations of fibronectin-like-immunoreactive fibroblasts and S-100-like-immunoreactive glia in the dissociated superior cervical ganglion cultures. However, only a subpopulation of fibronectin-like-immunoreactive fibroblasts possessed atrial natriuretic peptide binding sites in the explant culture preparations. No atrial natriuretic peptide-like-immunoreactive cells were present in either culture. The distribution of autoradiographic grains over individual cell surfaces in culture was uniform, but there were distinct differences in the density of labelling of single cells of the same type. This apparent variation in the number of binding sites on glial cells and fibroblasts in culture did not seem to be related to the morphology of the cells or the surrounding cell types. No sympathetic neurones were labelled with autoradiographic grains in either the dissociated or explant culture preparations. However, the presence of atrial natriuretic peptide binding sites on non-neuronal cells of sympathetic ganglia in culture may be linked to the relationship between atrial natriuretic peptide and the sympathetic nervous system.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Atlas SA, Kleinert HD, Camargo MJ, Januszewicz A, Sealey JE, Laragh JH, Schilling JW, Lewicki JA, Johnson LK, Maack T (1984) Purification, sequencing and synthesis of natriuretic and vasoactive rat atrial peptide. Nature 309:717–719

    Google Scholar 

  • Bloch KD, Seidman JG, Naftilan JD, Fallon JT, Seidman CE (1986) Neonatal atria and ventricles secrete atrial natriuretic factor via tissue-specific secretory pathways. Cell 47:695–702

    Google Scholar 

  • Bornstein MB (1958) Reconstituted rat-tail collagen used as a substrate for tissue cultures on coverslips in Maximow slides and roller tubes. Lab Invest 7:134–140

    Google Scholar 

  • Brown J, Polak JM, Power RF, Salas SP, Singleton A, Wharton J (1988) Differential distribution of subtypes of specific binding sites for alpha-atrial natriuretic peptide (αANP) in the rat kidney. J Physiol (Lond) 403:21P

    Google Scholar 

  • Buckley NJ, Saffrey MJ, Hassall CJS, Burnstock G (1988) Localization of muscarinic receptors on peptide-containing neurones of the guinea pig myenteric plexus in culture. Brain Res 445:152–156

    Google Scholar 

  • Burgess SK, McCarthy KD (1985) Autoradiographic quantitation of β-adrenergic receptors on neural cells in primary cultures. I. Pharmacological studies of [125I] pindolol binding on individual astroglial cells. Brain Res 335:1–9

    Google Scholar 

  • Chamley JH, Mark GE, Campbell GR, Burnstock G (1972a) Sympathetic ganglia in culture. I. Neurons. Z Zellforsch 135:287–314

    Google Scholar 

  • Chamley JH, Mark GE, Burnstock G (1972b) Sympathetic ganglia in culture. II. Accessory cells. Z Zellforsch 135:315–327

    Google Scholar 

  • Chinkers M, Garbers DL, Chang M-S, Lowe DG, Chin H, Goeddel DV, Schultz S (1989) A membrane form of guanylate cyclase is an atrial natriuretic peptide receptor. Nature 338:78–83

    Google Scholar 

  • Cramb G, Banks R, Rugg EL, Aiton JF (1987) Actions of atrial natriuretic peptide (ANP) on cyclic nucleotide concentrations and phosphatidylinositol turnover in ventricular myocytes. Biochem Biophys Res Commun 148:962–970

    Google Scholar 

  • Currie MG, Geller DM, Cole BR, Siegel NR, Fok KF, Adams SP, Eubanks SR, Galluppi GR, Needleman P (1984) Purification and sequence analysis of bioactive atrial peptides (atriopeptins). Science 223:67–69

    Google Scholar 

  • Debinski W, Gutkowska J, Kuchel O, Racz K, Buu NT, Cantin M, Genest J (1987a) Presence of an atrial natriuretic factor-like peptide in the rat superior cervical ganglia. Neuroendocrinology 46:236–240

    Google Scholar 

  • Debinski W, Kuchel O, Buu NT, Nemer M, Cantin M, Genest J (1987b) Atrial natriuretic factor in peripheral sympathetic ganglia originates in preganglionic nerve fibres. Clin Res 35:273A

    Google Scholar 

  • Debinski W, Kuchel O, Buu NT, Cantin M, Genest J (1987c) Atrial natriuretic factor partially inhibits the stimulated catecholamine synthesis in superior cervical ganglia of the rat. Neurosci Lett 77:92–96

    Google Scholar 

  • Freschi JE (1982) Effect of serum-free medium on growth and differentiation of sympathetic neurons in culture. Dev Brain Res 4:455–464

    Google Scholar 

  • Gardner DG, Vlasuk GP, Baxter JD, Fiddes JC, Lewicki JA (1987) Identification of atrial natriuretic factor gene transcripts in the central nervous system of the rat. Proc Natl Acad Sci USA 84:2175–2179

    Google Scholar 

  • Grammer RT, Fukumi H, Inagami T, Misono KS (1983) Rat atrial natriuretic factor: purification and vasorelaxant activity. Biochem Biophys Res Commun 116:696–703

    Google Scholar 

  • Gulbenkian S, Wharton J, Polak JM (1987) The visualisation of cardiovascular innervation in the guinea pig using an antiserum to protein gene product 9.5 (PGP 9.5). J Auton Nerv Syst 18:235–247

    Google Scholar 

  • Gutkind JS, Kurihara M, Castren E, Saavedra JM (1987) Atrial natriuretic peptide receptors in sympathetic ganglia: biochemical response and alterations in genetically hypertensive rats. Biochem Biophys Res Commun 149:65–72

    Google Scholar 

  • Hacker GW, Polak JM, Springall DR, Ballesta J, Cadieux A, Gu J, Trojanowski JQ, Dahl D, Marangos PJ (1985) Antibodies to neurofilament protein and other brain proteins reveal the innervation of peripheral organs. Histochemistry 82:581–593

    Google Scholar 

  • Hamid Q, Wharton J, Terenghi G, Hassall CJS, Aimi J, Taylor KM, Nakazato H, Dixon JE, Burnstock G, Polak JM (1987) Localization 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 

  • Haskins JT, Zingaro GJ, Lappe RW (1986) Rat atriopeptin III alters hypothalamic neuronal activity. Neurosci Lett 67:279–284

    Google Scholar 

  • Hassall CJS, Buckley NJ, Burnstock G (1987) Autoradiographic localisation of muscarinic receptors on guinea pig intracardiac neurones and atrial myocytes in culture. Neurosci Lett 74:145–150

    Google Scholar 

  • Hassall CJS, Wharton J, Gulbenkian S, Anderson JV, Frater J, Bailey DJ, Merighi A, Bloom SR, Polak JM, Burnstock G (1988) Ventricular and atrial myocytes on newborn rats synthesise and secrete atrial natriuretic peptide in culture: light-and electron-microscopical localisation and chromatographic examination of stored and secreted molecular forms. Cell Tissue Res 251:161–169

    Google Scholar 

  • Hassall CJS, Allen TGJ, Pittam BS, Burnstock G (1989) The use of cell and tissue culture techniques in the study of regulatory peptides. In: Polak JM (ed) Regulatory peptides. Birkhäuser, Basel, pp 113–136

    Google Scholar 

  • Hösli E, Hösli L (1987) Autoradiographic localization of binding sites for3H-serotonin and3H-ketanserin on neurones and astrocytes of cultured rat brain stem and spinal cord. Exp Brain Res 65:486–490

    Google Scholar 

  • Inagaki S, Kubota Y, Kito S, Kangawa K, Matsuo H (1986) Immunoreactive atrial natriuretic polypeptides in the adrenal medulla and sympathetic ganglia. Regul Pept 15:249–260

    Google Scholar 

  • James S, Burnstock G (1988) Autoradiographic localization of binding sites for125I-substance P on neurones from cultured rat superior cervical ganglion. Brain Res 458:205–211

    Google Scholar 

  • Kawata M, Ueda S, Nakao K, Morii N, Kiso Y, Imura H, Sano Y (1985) Immunohistochemical demonstration of α-atrial natriuretic polypeptide-containing neurons in the rat brain. Histochemistry 83:1–3

    Google Scholar 

  • Kuchel O, Debinski W, Racz K, Buu NT, Garcia R, Cusson JR, Larochelle P, Cantin M, Genest J (1987) An emerging relationship between peripheral sympathetic nervous activity and atrial natriuretic factor. Life Sci 40:1545–1551

    Google Scholar 

  • Leitman DC, Murad F (1986) Comparison of binding and cyclic GMP accumulation by atrial natriuretic peptides in endothelial cells. Biochem Biophys Acta 885:74–79

    Google Scholar 

  • Leitman DC, Agnost VL, Tuan JJ, Andresen JW, Murad F (1987) Atrial natriuretic factor and sodium nitroprusside increase cyclic GMP in cultured rat lung fibroblasts by activating different forms of guanylate cyclase. Biochem J 244:69–74

    Google Scholar 

  • Maack T, Suzuki M, Almeida FA, Nussenzveig D, Scarborough RM, McEnroe GA, Lewicki JA (1987) Physiological role of silent receptors of atrial natriuretic factor. Science 238:675–678

    Google Scholar 

  • Mantyh CR, Kruger L, Brecha NC, Mantyh PW (1986) Localization of specific binding sites for atrial natriuretic factor in peripheral tissues of the guinea pig, rat and human. Hypertension 8:712–721

    Google Scholar 

  • Marrion NV, Smart TG, Brown DA (1987) Membrane currents in adult rat superior cervical ganglia in dissociated tissue culture. Neurosci Lett 77:55–60

    Google Scholar 

  • Mirsky R, Jessen KR (1986) Glial cells of the brain and peripheral nervous system: some new perspectives. Rec Adv Neuropathol 3:1–24

    Google Scholar 

  • Morii M, Nakao K, Sugawara A, Sakamoto M, Suda M, Shimokura M, Kiso Y, Kihara M, Yamori Y, Imura H (1985) Occurrence of atrial natriuretic polypeptide in brain. Biochem Biophys Res Commun 127:413–419

    Google Scholar 

  • Napier MA, Vandlen RL, Albers-Schonberg G, Nutt RF, Brady S, Lyle T, Winquist R, Faison EP, Heinel LA, Blaine EH (1984) Specific membrane receptors for atrial natriuretic factor in renal and vascular tissues. Proc Natl Acad Sci USA 81:5946–5950

    Google Scholar 

  • Niwa M, Shigematsu K, Kurihara M, Kataoka Y, Maeda T, Nakao K, Imura H, Matsuo H, Tsuchiyama H, Ozaki M (1988) Receptor autoradiographic evidence of specific brain natriuretic peptide binding sites in the porcine subfornical organ. Neurosci Lett 95:113–118

    Google Scholar 

  • Nussenzveig DR, Owada A, Scarborough R, Arfsten A, Maack T (1988) Biological (B) and clearance (C) receptors of ANF in cultured mesangial cells and fibroblasts. Am J Hypertens 1:Abs 5003

  • O'Lague PH, Potter DD, Furshpan EJ (1978) Studies on rat sympathetic neurons developing in culture. I. Growth characteristics and electrophysiological properties. Dev Biol 67:384–403

    Google Scholar 

  • Papka RE, Traurig HH, Wekstein M (1985) Localization of peptides in nerve terminals in the paracervical ganglion of the rat by light and electron microscopic immunohistochemistry: enkephalin and atrial natriuretic factor. Neurosci Lett 61:285–290

    Google Scholar 

  • Quirion R, Dalpe M, De Léan A, Gutkowska J, Cantin M, Genest J (1984) Atrial natriuretic factor (ANF) binding sites in brain and related structures. Peptides 5:1167–1172

    Google Scholar 

  • Raff MC, Fields KL, Hakomori S, Mirsky R, Pruss RM, Winter J (1979) Cell-type-specific markers for distinguishing and studying neurons and the major classes of glial cells in culture. Brain Res 174:283–308

    Google Scholar 

  • Reiser G, Hopp H-P, Hamprecht B (1987) Atrial natriuretic hormones induce membrane potential responses in cultured rat glioma. Brain Res 402:164–167

    Google Scholar 

  • Saper CB, Hurley KM, Moga MM, Holmes HR, Adams SA, Leahy KM, Needleman P (1989) Brain natriuretic peptides: differential localization of a new family of neuropeptides. Neurosci Lett 96:29–34

    Google Scholar 

  • Sasaki A, Kida O, Kangawa K, Matsuo H, Tanaka K (1986) Involvement of sympathetic nerves in cardiosuppressive effects of α-human atrial natriuretic polypeptide (α-ANP) in anesthetized rats. Eur J Pharmacol 120:345–349

    Google Scholar 

  • Scarborough RM, Schenk DB, McEnroe GA, Arfsten A, Kang L-L, Schwartz K, Lewicki JA (1986) Truncated atrial natriuretic peptide analogs. Comparison between receptor binding and stimulation of cyclic GMP accumulation in cultured vascular smooth muscle cells. J Biol Chem 261:12960–12964

    Google Scholar 

  • Simonnet G, Legendre P, Laribi C, Allard M, Vincent JD (1988) Location of angiotensin II binding sites on neuronal and glial cells of cultured mouse spinal cord: an autoradiographic study. Brain Res 443:403–406

    Google Scholar 

  • Song D-L, Kohse KP, Murad F (1988) Brain natriuretic factor. Augmentation of cellular cyclic GMP, activation of particulate guanylate cyclase and receptor binding. FEBS Lett 232:125–129

    Google Scholar 

  • Sudoh T, Kangawa K, Minamino N, Matsuo H (1988) A new natriuretic peptide in porcine brain. Nature 332:78–81

    Google Scholar 

  • Van Calker D, Hamprecht B (1980) Effects of neurohormones on glial cells. Adv Cell Neurobiol 1:31–67

    Google Scholar 

  • Varon SS, Bunge RP (1978) Trophic mechanisms in the peripheral nervous system. Annu Rev Neurosci 1:327–361

    Google Scholar 

  • Winquist RJ, Faison EP, Waldman SA, Schwartz K, Murad F, Rapoport RM (1984) Atrial natriuretic factor elicits an endothelium-independent relaxation and activates particulate guanylate cyclase. Proc Natl Acad Sci USA 81:7661–7664

    Google Scholar 

  • Yamada SS, Yamada K, Willingham MC (1980) Intracellular localization of fibronectin by immunoelectron microscopy. J Histochem Cytochem 28:953–960

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Anatomy and Developmental Biology, University College London, London, United Kingdom

    S. James, C. J. S. Hassall & G. Burnstock

  2. Centre for Neuroscience, University College London, London, United Kingdom

    S. James, C. J. S. Hassall & G. Burnstock

  3. Department of Histochemistry, Royal Postgraduate Medical School, London, United Kingdom

    J. M. Polak

Authors
  1. S. James
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. C. J. S. Hassall
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J. M. Polak
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. G. Burnstock
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

James, S., Hassall, C.J.S., Polak, J.M. et al. Visualisation of specific binding sites for atrial natriuretic peptide on non-neuronal cells of cultured rat sympathetic ganglia. Cell Tissue Res. 259, 129–137 (1990). https://doi.org/10.1007/BF00571437

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00571437

Key words

Search

Navigation