Cell and Tissue Research

, Volume 261, Issue 2, pp 339–345 | Cite as

Immunohistochemical identification of calcitonin gene-related peptide and substance P in nerves of the bovine parathyroid gland

  • Shane T. Mortimer
  • David A. Hanley
  • William K. Stell


Although peptide neurotransmitters have been shown to modulate hormone secretion in many glands, there are very few studies of neurotransmitters in the parathyroid gland. Bovine parathyroid glands were collected at a local abattoir, fixed with paraformaldehyde, sectioned using a cryostat, and stained by indirect immunohistochemistry for calcitonin gene-related peptide and substance P. We were able to positively identify both neuropeptides. Nerve fibres containing calcitonin gene-related peptide and substance P were identified in contact with the tunica media of arteries and arterioles and dispersed throughout the stroma of the gland. While many of the fibres encircled parenchymal lobules, no intimate contact with the peripheral chief cells was observed. All immunoreactive fibres were found to contain both neuropeptides. Since calcitonin gene-related peptide and substance P are vasodilators, they may increase blood flow within the gland. In addition, the neuropeptides may diffuse from perilobular nerve fibres into the parenchyma, thereby modulating secretion of parathyroid hormone.

Key words

Parathyroid glands Neuropeptide colocalization Calcitonin gene-related peptide (CGRP) Substance P Immunohistochemistry Cow 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Ahrén B, Alumets J, Ericsson M, Fahrenkrug J, Fahrenkrug L, Håkanson R, Hedner P, Lorén I, Melander A, Rerup C, Sundler F (1980) VIP occurs in intrathyroidal nerves and stimulates thyroid hormone secretion. Nature 287:343–345Google Scholar
  2. Ahrén B, Grunditz T, Ekman R, Håkanson R, Sundler F, Uddman R (1983) Neuropeptides in the thyroid gland: distribution of substance P and gastrin/cholecystokinin and their effects on the secretion of iodothyronine and calcitonin. Endocrinology 113:379–384Google Scholar
  3. Altenähr E (1971) Electron microscopical evidence for innervation of chief cells in human parathyroid gland. Experientia 27:1077Google Scholar
  4. Atwal OS (1981) Myelinated nerve fibres in the parathyroid gland of the dog: a light and electron-microscopic study. Acta Anat 109:3–12Google Scholar
  5. Brain SD, Williams TJ, Tippins JR, Morris HR, MacIntyre I (1985) Calcitonin gene-related peptide is a potent vasodilator. Nature 313:54–56Google Scholar
  6. Brown EM, Carroll RJ, Aurbach GD (1977a) Dopaminergic stimulation of cyclic AMP accumulation and parathyroid hormone release from dispersed bovine parathyroid cells. Proc Natl Acad Sci USA 74:4210–4213Google Scholar
  7. Brown EM, Hurwitz S, Aurbach GD (1977b) Beta-adrenergic stimulation of cyclic AMP content and parathyroid hormone release from isolated bovine parathyroid cells. Endocrinology 100:1696–1702Google Scholar
  8. Brown EM, Hurwitz S, Woodard CJ, Aurbach GD (1977c) Direct identification of beta-adrenergic receptors on isolated bovine parathyroid cells. Endocrinology 100:1703–1709Google Scholar
  9. Brown EM, Watson EJ, Leombruno R, Underwood RH (1983) Extracellular calcium is not necessary for acute, low calciumor dopamine-stimulated PTH secretion in dispersed bovine parathyroid cells. Metabolism 32:1038–1044Google Scholar
  10. Brown EM, Redgrave J, Thatcher J (1984) Effect of the phorbol ester TPA on PTH secretion. FEBS Lett 175:72–75Google Scholar
  11. Capen CC, Koestner A, Cole CR (1965) The ultrastructure and histochemistry of normal parathyroid glands of pregnant and nonpregnant cows. Lab Invest 14:1673–1690Google Scholar
  12. Carlton SM, McNeill DL, Chung K, Coggeshall RE (1987) A light and electron microscopic level analysis of calcitonin gene-related peptide (CGRP) in the spinal cord of the primate: an immunohistochemical study. Neurosci Lett 82:145–150Google Scholar
  13. Cuello AC, Galfre G, Milstein C (1979) Detection of substance P in the central nervous system by a monoclonal antibody. Proc Natl Acad Sci USA 76:3532–3536Google Scholar
  14. Deftos LJ, Parthemore JG (1974) Secretion of parathyroid hormone in patients with medullary thyroid carcinoma. J Clin Invest 54:416–420Google Scholar
  15. Edbrooke MR, Parker D, McVey JH, Riley JH, Sorenson GD, Pettengill OS, Craig RK (1985) Expression of the human calcitonin/CGRP gene in lung and thyroid carcinoma. EMBO J 4:715–724Google Scholar
  16. Epstein PA, Prentki M, Attie MF (1985) Modulation of intracellular Ca2+ in the parathyroid cell. FEBS Lett 188:141–144Google Scholar
  17. Fisher LA, Kikkawa DO, Rivier JE, Amara SG, Evans RM, Rosenfeld MG, Vale WW, Brown MR (1983) Stimulation of noradrenergic sympathetic outflow by calcitonin gene-related peptide. Nature 305:534–536Google Scholar
  18. Gibbins IL, Furness JB, Costa M, MacIntyre I, Hillyard CJ, Girgis S (1985) Co-localization of calcitonin gene-related peptide-like immunoreactivity with substance P in cutaneous, vascular and visceral sensory neurons of guinea pigs. Neurosci Lett 57:125–130Google Scholar
  19. Gibson SJ, Polak JM, Bloom SR, Sabate IM, Mulderry PM, Ghatel MA, McGregor GP, Morrison JFB, Kelly JS, Evans RM, Rosenfeld MG (1984) Calcitonin gene-related peptide immunore-activity in the spinal cord of man and of eight other species. J Neurosci 4:3101–3111Google Scholar
  20. Grunditz T, Ekman R, Håkanson R, Rerup C, Sundler F, Uddman R (1986) Calcitonin gene-related peptide in thyroid nerve fibres and C cells: effects on thyroid hormone secretion and response to hypercalcemia. Endocrinology 119:2313–2324Google Scholar
  21. Hanley DA, Wellings PG (1985) Dopamine-stimulated parathyroid hormone release in vitro: further evidence for a two-pool model of parathyroid hormone secretion. Can J Physiol Pharmacol 63:1139–1144Google Scholar
  22. Hanley DA, Takatsuki K, Birnbaumer ME, Schneider AB, Sherwood LM (1980a) In vitro perifusion for the study of parathyroid hormone secretion: effects of extracellular calcium concentration and beta-adrenergic regulation on bovine parathyroid hormone secretion in vitro. Calcif Tissue Int 32:19–27Google Scholar
  23. Hanley MR, Lee CM, Jones LM, Michell RH (1980b) Similar effects of substance P and related peptides on salivation and on phosphatidylinositol turnover in rat salivary glands. Mol Pharmacol 18:78–83Google Scholar
  24. Hardebo JE, Suzuki N, Owman C (1989) Origins of substance P- and calcitonin gene-related peptide-containing nerves in the internal carotid artery of rat. Neurosci Lett 101:39–45Google Scholar
  25. Kruger L, Silverman JD, Mantyh PW, Sternini C, Brecha NC (1989) Peripheral patterns of calcitonin-gene-related peptide general somatic sensory innervation: cutaneous and deep terminations. J Comp Neurol 280:291–302Google Scholar
  26. Lee Y, Kawai Y, Shiosaka S, Takami K, Kiyama H, Hillyard CJ, Girgis S, MacIntyre I, Emson PC, Tohyama M (1985a) Coexistence of calcitonin gene-related peptide and substance P-like peptide in single cells of the trigeminal ganglion of the rat: immunohistochemical analysis. Brain Res 330:194–196Google Scholar
  27. Lee Y, Takami K, Kawai Y, Girgis S, Hillyard CJ, MacIntyre I, Emson PC, Tohyama M (1985b) Distribution of calcitonin gene-related peptide in the rat peripheral nervous system with reference to its coexistence with substance P. Neuropeptides 15:1227–1237Google Scholar
  28. Lorén I, Emson PC, Fahrenkrug J, Björklund A, Alumets J, Håkanson R, Sundler F (1979) Distribution of vasoactive intestinal polypeptide in the rat and mouse brain. Neuroscience 4:1953–1976Google Scholar
  29. Lundberg JM, Höfelt T, Hemsén A, Theodorsson-Norheim E, Pernow J, Hamberger B, Goldstein M (1986) Neuropeptide Y-like immunoreactivity in adrenaline cells of adrenal medulla and in tumors and plasma of pheochromocytoma patients. Regul Rep 13:169–182Google Scholar
  30. MacGregor RR, Sarras Jr MP, Houle A, Cohn DV (1983) Primary monolayer cell culture of bovine parathyroids: effects of calcium, isoproterenol and growth factors. Mol Cell Endocrinol 30:313–328Google Scholar
  31. Mantyh PW, Pinnock RD, Downes CP, Goedert M, Hunt SP (1984) Correlation between inositol phospholipid hydrolysis and substance P receptors in rat CNS. Nature 309:795–797Google Scholar
  32. Membreño L, Chen T, Woddley S, Gagucas R (1989) The effects of protein kinase-C agonists on parathyroid hormone release and intracellular free Ca2+ in bovine parathyroid cells. Endocrinology 124:789–797Google Scholar
  33. Mikhail Y (1971) Intrinsic nerve supply of the thyroid and parathyroid glands. Acta Anat 80:152–159Google Scholar
  34. Morel G, Besson J, Rosselin G, Dubois PM (1982) Ultrastructural evidence for endogenous vasoactive intestinal peptide-like immunoreactivity in the pituitary gland. Neuroendocrinology 34:85–89Google Scholar
  35. Morrissey JJ (1988) Effect of phorbol myristate acetate on secretion of parathyroid hormone. Am J Physiol 254:E63-E70Google Scholar
  36. Nemeth EF, Wallace J, Scarpa A (1986) Stimulus-secretion coupling in bovine parathyroid cells. J Biol Chem 261:2668–2674Google Scholar
  37. Norberg KA, Persson B, Granberg PO (1975) Adrenergic innervation of the human parathyroid glands. Acta Chir Scand 141:319–322Google Scholar
  38. Pettersson M, Ahrén B, Böttcher G, Sundler F (1986) Calcitonin gene-related peptide: occurrence in pancreatic islets in the mouse and the rat and inhibition of insulin secretion in the mouse. Endocrinology 119:865–869Google Scholar
  39. Raybuck HE (1952) The innervation of the parathyroid glands. Anat Rec 112:117–123Google Scholar
  40. Rhinchart DA (1912) The nerves of the thyroid and parathyroid bodies. Am J Anat 13:91–102Google Scholar
  41. Rosenfeld MG, Mermod JJ, Amara SG, Swanson LW, Sawchenko PE, Rivier J, Vale WW, Evans RM (1983) Production of a novel neuropeptide encoded by the calcitonin gene via tissue-specific RNA processing. Nature 304:129–135Google Scholar
  42. Sabate MI, Stolarsky LS, Polak JM, Bloom SR, Varndell IM, Ghatei MA, Evans RM, Rosenfeld MG (1985) Regulation of neuroendocrine gene expression by alternate RNA processing. J Biol Chem 260:2589–2592Google Scholar
  43. Skofitsch G, Jacobowitz DM (1985a) Calcitonin gene-related peptide: detailed immunohistochemical distribution in the central nervous system. Peptides 6:721–745Google Scholar
  44. Skofitsch G, Jacobowitz DM (1985b) Calcitonin gene-related peptide coexists with substance P in capsaicin sensitive neurons and sensory ganglia of the rat. Peptides 6:747–754Google Scholar
  45. Sternini C, Brecha N (1986) Immunocytochemical identification of islet cells and nerve fibres containing calcitonin gene-related peptide-like immunoreactivity in the rat pancreas. Gastroenterology 90:1155–1163Google Scholar
  46. Stone RA, McGlinn AM (1988) Calcitonin gene-related peptide immunoreactive nerves in human and rhesus monkey eyes. Invest Ophthalmol Vis Sci 29:305–310Google Scholar
  47. Tippins JR, Morris HR, Panico M, Etienne T, Bevis P, Girgis S, MacIntrye I, Azria M, Attinger M (1984) The myotropic and plasma-calcium modulating effects of calcitonin gene-related peptide (CGRP). Neuropeptides 4:425–434Google Scholar
  48. Unsicker K (1971) On the innervation of mammalian endocrine glands (anterior pituitary and parathyroids). Z Mikrosk Anat Forsch 121:283–291Google Scholar
  49. Varndell IM, Polak JM, Allen JM, Terenghi G, Bloom SR (1984) Neuropeptide tyrosine (NPY) immunoreactivity in norepinephrine-containing cells and nerves of the mammalian adrenal gland. Endocrinology 114:1460–1462Google Scholar
  50. Watson SP, Downes CP (1983) Substance P induced hydrolysis of inositol phospholipids in guinea-pig ileum and rat hypothalamus. Eur J Pharmacol 93:245–253Google Scholar
  51. Wideman Jr RF (1980) Innervation of the parathryoid in the European starling (Sturnus vulgaris). J Morphol 166:65–80Google Scholar
  52. Wiesenfeld-Hallin Z, Hökfelt T, Lundberg JM, Forssmann WG, Reinecke M, Tschopp FA, Fischer JA (1984) Immunoreactive calcitonin gene-related peptide and substance P coexist in sensory neurons to the spinal cord and interact in spinal behavioral responses of the rat. Neurosci Lett 52:199–204Google Scholar
  53. Yeghiayan E, Rojo-Ortega JM, Genest J (1972) Parathyroid vessel innervation: an ultrastructural study. J Anat 112:137–142Google Scholar
  54. Zabel M, Biela-Jacek I, Surdyk J, Dietel M (1987) Studies on localization of calcitonin gene-related peptide (CGRP) in the thyroid-parathyroid complex. Virchows Arch 411A:569–573Google Scholar
  55. Zamboni L, De Martino C (1967) Buffered picric acid-formaldehyde: a new, rapid fixative for electronmicroscopy. J Cell Biol 35:148AGoogle Scholar
  56. Zawistowski S (1966) Histochemical studies on the adrenergic innervation of the parathyroid gland of albino rat. Z Mikrosk Anat Forsch 74:39–45Google Scholar

Copyright information

© Springer-Verlag 1990

Authors and Affiliations

  • Shane T. Mortimer
    • 1
  • David A. Hanley
    • 1
  • William K. Stell
    • 2
  1. 1.Department of Medicine, Faculty of MedicineThe University of CalgaryCalgaryCanada
  2. 2.Department of Anatomy, Faculty of MedicineThe University of CalgaryCalgaryCanada
  3. 3.Endocrine Research Group, Department of MedicineThe University of Calgary, Health Science CentreCalgaryCanada

Personalised recommendations