Anatomy and Embryology

, Volume 188, Issue 3, pp 219–237 | Cite as

Innervation of the dura mater encephali of cat and rat: ultrastructure and calcitonin gene-related peptide-like and substance P-like immunoreactivity

  • Karl Meßlinger
  • Ulrike Hanesch
  • Matthias Baumgärtel
  • Brigitte Trost
  • Robert F. Schmidt
Original Articles


Ultrastructural, immunocytochemical, and immunoelectron microscopical examinations are reported that describe the morphology of putative sensory nerve endings in the dura mater encephali of the rat and the cat. Morphometrical measurements and reconstructions showed that in the cat the mean diameter of axons, the bare area of axolemma, and the content of mitochondria and vesicles are highly variable in dural nerve endings. Nerve fibers with a high volume density of mitochondria are thought to be sensory, while nerve fibers containing many small vesicles are considered autonomic. There is, however, a broad overlap of mitochondria-rich and vesicle-rich nerve fibers in the dura, so that discrimination between sensory and autonomic endings by these characteristics frequently fails. Whole-mount preparations treated cytochemically for detection of substance P- and calcitonin gene-related peptide-like immunoreactivity in the rat and the cat showed a network of immunopositive nerve fibers in the vicinity of dural blood vessels. Most of these peptidergic and probably sensory nerve fibers were found terminating in the dural connective tissue far from vessels. Calcitonin gene-related peptide-positive nerve fibers were much more abundant than substance P-positive fibers. Immunoelectron microscopic preparations revealed that calcitonin gene-related peptide- and substance P-like immunoreactivity is found in a small proportion of generally thin unmyelinated nerve fibers. These proportions were very similar in the rat and the cat. Summarizing the recent literature, the morphological characteristics of putative sensory nerve fibers in the dura mater are discussed in relation to their possible functional significance for neurogenic inflammation and nociception.

Key words

Sensory innervation Meninges Neuropeptides Neurogenic inflammation Headache 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Amenta F, Sancesario G, Ferrante F, Cavallotti C (1980) Acetylcholinesterase-containing nerve fibers in the dura mater of guinea pig, mouse, and rat. J Neural Transm 47:237–242Google Scholar
  2. Andres KH, von Düring M (1973) Morphology of cutaneous receptors. In: Iggo A (ed) Handbook of sensory physiology, vol 2. Springer, Berlin Heidelberg New York, pp 3–28Google Scholar
  3. Andres KH, von Düring M, Schmidt RF (1985) Sensory innervation of the Achilles tendon by group III and IV afferent fibres. Anat Embryol 172:145–156Google Scholar
  4. Andres KH, von Düring M, Muszynski K, Schmidt RF (1987) Nerve fibres and their terminals of the dura mater encephali of the rat. Anat Embryol 175:289–301Google Scholar
  5. Burnstock G (1981) Ultrastructural identification of neurotransmitters. Scand J Gastroenterol 16 [Suppl 70]: 1–9Google Scholar
  6. Buzzi MG, Carter WB, Shimizu T, Heath III H, Moskowitz MA (1991) Dihydroergotamine and sumatriptan attenuate levels of CGRP in plasma in rat superior sagittal sinus during electrical stimulation of the trigeminal ganglion. Neuropharmacology 30:1193–1200Google Scholar
  7. Byers MR (1985) Sensory innervation of periodontal ligament of rat molars consists of unencapsulated Ruffini-like mechanoreceptors and free nerve endings. J Comp Neurol 231:500–518Google Scholar
  8. Cameron AA, Leah JH, Snow PJ (1988) The coexistence of peptides in feline sensory neurons. Neuroscience 27:969–979Google Scholar
  9. Davis KD, Dostrovsky JO (1986) Activation of trigeminal brainstem nociceptive neurons by dural artery stimulation. Pain 25:395–401Google Scholar
  10. Dimitriadou V, Buzzi MG, Moskowitz MA, Theoharides TC (1991) Trigeminal sensory fiber stimulation induces morphological changes reflecting secretion in rat dura mater mast cells. Neuroscience 44:97–112Google Scholar
  11. Dimlich RVW, Keller JT, Strauss TA, Fritts MJ (1991) Linear arrays of homogenous mast cells in the dura mater of the rat. J Neurocytol 20:485–503Google Scholar
  12. Dixon JS, Gilpin CJ (1987) Presumptive sensory axons of the human urinary bladder: a fine structural study. J Anat 151:199–207Google Scholar
  13. Dostrovsky JO, Davis KD, Kawakita K (1991) Central mechanisms of vascular headache. Can J Physiol Pharmacol 69:652–658Google Scholar
  14. Dowgjallo N (1929) Über die Nerven der harten Hirnhaut des Menschen und einiger Säuger. Z Ges Anat 89:453–466Google Scholar
  15. Düring M von, Bauersachs M, Böhmer B, Veh RW, Andres KH (1990) Neuropeptide Y- and substance P-like immunoreactive nerve fibers in the rat dura mater encephali. Anat Embryol 182:363–373Google Scholar
  16. Edvinsson L, Uddman R (1981) Adrenergic, cholinergic and peptidergic nerve fibres in dura mater — involvement in headache? Cephalalgia 1:175–179Google Scholar
  17. Edvinsson L, Rosendahl-Helgesen S, Uddman R (1983) Substance P: localization, concentration and release in cerebral arteries, choroid plexus and dura mater. Cell Tissue Res 234:1–7Google Scholar
  18. Edvinsson L, Fredholm BB, Hamel E, Jansen I, Verrecchia C (1985) Perivascular peptides relax cerebral arteries concomitant with stimulation of cyclic adenosine monophosphate accumulation or release of an endothelium-derived relaxing factor in the cat. Neurosci Lett 58:213–217Google Scholar
  19. Edvinsson L, Brodin E, Jansen I, Uddman R (1988) Neurokinin A in cerebral vessels: characterization, localization and effects in vitro. Regul Peptides 20:181–197Google Scholar
  20. Edwards RM, Stack EJ, Trizna W (1991) Calcitonin gene-related peptide stimulates adenylate cyclase and relaxes intracerebral arterioles. J Pharmacol Exp Ther 257:1020–1024Google Scholar
  21. Fried G, Terenius L, Hökfelt T, Goldstein M (1985) Evidence for differential localization of noradrenaline and neuropeptide Y in neuronal storage vesicles isolated from rat vas deferens. J Neurosci 5:450–458Google Scholar
  22. Fried G, Franck J, Brodin E, Born W, Fischer JA, Hiort W, Hökfelt T (1989) Evidence for differential storage of calcitonin generelated peptide, substance P and serotonin in synaptosomal vesicles of rat spinal cord. Brain Res 499:315–324Google Scholar
  23. Geppetti P, del Bianco E, Santicolli P, Maggi CA, Tramontana M, Sicuteri F (1989) Neuropeptide release from sensory fibres of guinea pig cerebral venous sinuses and dorsal spinal cord: relevance for headache study. Cephalalgia 9 [Suppl 10]: 15–16Google Scholar
  24. Gibbins JL, 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 neurones of guinea pigs. Neurosci Lett 57:125–130Google Scholar
  25. Goadsby PJ, Edvinsson L, Ekman R (1988) Release of vasoactive peptides in the extracerebral circulation of humans and the cat during activation of the trigeminovascular system. Ann Neurol 123:193–196Google Scholar
  26. Grigg P, Schaible H-G, Schmidt RF (1986) Mechanical sensitivity of group III and IV afferents from posterior articular nerve in normal and inflamed cat knee. J Neurophysiol 55:635–643Google Scholar
  27. Gulbenkian S, Merighi A, Wharton J, Varndell IM, Polak JM (1986) Ultrastructural evidence for the coexistence of calcitonin gene-related peptide and substance P in secretory vesicles of peripheral nerves in the guinea pig. J Neurocytol 15:535–542Google Scholar
  28. Häbler H-J, Jänig W, Koltzenburg M (1990) Activation of unmyelinated afferents by mechanical stimuli and inflammation of the urinary bladder. J Physiol (Lond) 425:545–563Google Scholar
  29. Handwerker HO, Kilo S, Reeh PW (1991) Unresponsive afferent nerve fibres in the sural nerve of the rat. J Physiol 435:229–242Google Scholar
  30. Hanesch U, Heppelmann B, Schmidt RF (1991a) Substance P- and calcitonin gene-related peptide immunoreactivity in primary afferent neurons of the cat's knee joint. Neuroscience 45:185–193Google Scholar
  31. Hanesch U, Meßlinger K, Schmidt RF (1991b) CGRP-immunoreactive nerve fibers in the dura mater encephali of cat and rat. Pflügers Arch 418:R34Google Scholar
  32. 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
  33. Heppelmann B, Meßlinger K, Schmidt RF (1989) Serial sectioning, electron microscopy, and three-dimensional reconstruction of fine nerve fibers and other extended objects. J Microscopy 156:163–172Google Scholar
  34. Hoyes AD, Barber P (1976) Ultrastructure of the corneal nerves in the rat. Cell Tissue Res 172:133–144Google Scholar
  35. Iwayama T, Furness JB, Burnstock G (1970) Dual adrenergic and cholinergic innervation of the cerebral arteries of the rat. Circulation Res 26:635–646Google Scholar
  36. Jansen I, Alafaci C, McCulloch J, Uddman R, Edvinsson L (1991) Tachykinins (substance P, neurokinin A, neuropeptide K, and neurokinin B) in the cerebral circulation: vasomotor responses in vitro and in situ. J Cereb Blood Flow Metab 11:567–575Google Scholar
  37. Kawatani M, Nagel J, de Groat WC (1986) Identification of neuropeptides in pelvic and pudendal afferent pathways to the sacral spinal cord of the cat. J Comp Neurol 249:117–132Google Scholar
  38. Keller JT, Marfurt CF (1991) Peptidergic and serotoninergic innervation of the rat dura mater. J Comp Neurol 309:515–534Google Scholar
  39. Keller JT, Saunders MC, Beduk A, Jollis JG (1985) Innervation of the posterior fossa dura of the cat. Brain Res Bull 14:97–102Google Scholar
  40. Keller JT, Marfurt CF, Dimlich RVW, Tierney BE (1989) Sympathetic innervation of the supratentorial dura mater of the rat. J Comp Neurol 290:310–321Google Scholar
  41. Komuro T, Baluk P, Burnstock G (1982) An ultrastructural study of nerve profiles in the myenteric plexus of the rabbit colon. Neuroscience 7:295–305Google Scholar
  42. Langford LA, Coggeshall RE (1980) The use of potassium ferricyanide in neural fixation. Anat Rec 197:297–303Google Scholar
  43. Lee Y, Kawai Y, Shiosaka S, Takami K, Kiyama H, Hillyard CJ, Girgis S, Macintyre J, Emson PC, Tohyama M (1985) Co-existence 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
  44. Liu-Chen L-Y, Mayberg MR, Moskowitz MA (1983a) Immunohistochemical evidence for a substance P-containing trigemi-novascular pathway to pial arteries in cats. Brain Res 268:162–166Google Scholar
  45. Liu-Chen L-Y, Han DH, Moskowitz MA (1983b) Pia arachnoid contains substance P originating from trigeminal neurons. Neuroscience 9:803–808Google Scholar
  46. Liu-Chen L-Y, Liszczak TM, King JC, Moskowitz MA (1986) Immunoelectron microscopic study of substance P-containing fibers in feline cerebral arteries. Brain Res 369:12–20Google Scholar
  47. Markowitz S, Saito K, Moskowitz MA (1987) Neurogenically mediated leakage of plasma protein occurs from blood vessels in dura mater but not brain. J Neurosci 7:4129–4136Google Scholar
  48. Matsuyama T, Shiosaka S, Matsumoto M, Yoneda S, Kimura K, Abe H, Hayakawa T, Inoue H, Tohyama M (1983) Overall distribution of vasoactive intestinal polypeptide-containing nerves on the wall of cerebral arteries: an immunohistochemical study using whole-mounts. Neuroscience 10:89–96Google Scholar
  49. Matsuyama T, Matsumoto M, Shiosaka S, Hayakawa T, Yoneda S, Kimura K, Abe H, Tohyama M (1984) Dual innervation of substance P-containing neuron systems in the wall of the cerebral arteries. Brain Res 322:144–147Google Scholar
  50. Matsuyama T, Shiosaka S, Wanaka A, Yoneda S, Kimura K, Hayakawa T, Emson PC, Tohyama M (1985) Fine structure of peptidergic and catecholaminergic nerve fibers in the anterior cerebral artery and their interrelationship: an immunoelectron microscopic study. J Comp Neurol 235:268–276Google Scholar
  51. Mayberg MR, Nicholas TZ, Moskowitz MA (1984) Trigeminal projections to supratentorial pial and dural blood vessels in cats demonstrated by horseradish peroxidase histochemisty. J Comp Neurol 223:46–56Google Scholar
  52. Meßlinger K, Pawlak M, Trost B, Schmidt RF (1991) Quantitative Ultrastruktur feiner Nervenfaserendigungen in den Hirnhäuten der Katze. Anat Anz 172:55Google Scholar
  53. Meyer RA, Davis KD, Cohen RH, Treede R-D, Campbell JN (1991) Mechanically insensitive afferents (MIAs) in cutaneous nerves of monkey. Brain Res 561:252–262Google Scholar
  54. Molander C, Ygge J, Dalsgaard C-J (1987) Substance P-, somatostatin- and calcitonin gene-related peptide-like immunoreactivity and fluoride resistant acid phosphatase-activity in relation to retrogradely labelled cutaneous, muscular and visceral primary sensory neurones in the rat. Neurosci Lett 74:37–42Google Scholar
  55. Moskowitz MA (1984) The neurobiology of vascular head pain. Ann Neurol 16:157–168Google Scholar
  56. Moskowitz MA, Buzzi MG (1991) Neuroeffector functions of sensory fibres: implications for headache mechanisms and drug actions. J Neurol 238:S18-S22Google Scholar
  57. Moskowitz MA, Brody M, Liu-Chen L-Y (1983) In vitro release of immunoreactive substance P from putative afferent nerve endings in bovine pia arachnoid. Neuroscience 9:809–814Google Scholar
  58. Moskowitz MA, Buzzi MG, Sakas DE, Linnik MD (1989) Pain mechanisms underlying vascular headaches. Rev Neurol 145:181–193Google Scholar
  59. Nielsen KC, Owman C, Sporrong B (1971) Ultrastructure of the autonomic apparatus in the main pial arteries of rats and cats. Brain Res 27:25–32Google Scholar
  60. Nozaki K, Uemura Y, Okamoto S, Kikuchi H, Mizuno N (1990) Origins and distribution of cerebrovascular nerve fibers showing calcitonin gene-related peptide-like immunoreactivity in the major cerebral artery of the dog. J Comp Neurol 297:219–226Google Scholar
  61. O'Connor T, van der Kooy D (1986) Patterns of intracranial and extracranial projections of trigeminal ganglion cells. J Neurosci 6:2200–2207Google Scholar
  62. O'Connor T, van der Kooy D (1988) Enrichment of a vasoactive neuropeptide (calcitonin gene-related peptide) in the trigeminal sensory projection to the intracranial arteries. J Neurosci 8:2468–2476Google Scholar
  63. Oleson J, Edvinsson L (1991) Migraine: a research field matured for the basic neurosciences. Trends Neurol Sci 14:3–5Google Scholar
  64. Olgart L, Gazelius B, Brodin E, Nilsson G (1977) Release of substance P-like immunoreactivity from the dental pulp. Acta Physiol Scand 101:510–512Google Scholar
  65. Penfield W, McNaughton FL (1940) Dural headache and innervation of the dura mater. Arch Neurol Psychiatr 44:43–75Google Scholar
  66. Ray BS, Wolff HG (1940) Experimental studies on headache: pain sensitive structures of the head and their significance in headache. Arch Surg 1:813–856Google Scholar
  67. Schaible H-G, Schmidt RF (1983) Responses of fine medial articular nerve afferents to passive movements of knee joint. J Neurophysiol 49:1118–1126Google Scholar
  68. Schaible H-G, Schmidt RF (1988) Direct observation of the sensitization of articular afferents during an experimental arthritis. In: Dubner R, Gebhardt GF, Bond MR (eds) Proceedings of the Vth World Congress on Pain. Elsevier, Amsterdam New York Oxford, pp 44–50Google Scholar
  69. Steiger HJ, Meakin CJ (1984) The meningeal representation in the trigeminal ganglion — an experimental study in the cat. Headache 24:305–309Google Scholar
  70. Steiger HJ, Tew JM, Keller JT (1982) The sensory representation of the dura mater in the trigeminal ganglion of the cat. Neurosci Lett 31:231–236Google Scholar
  71. Stöhr P (1932) Nerves of the blood vessels, heart, meninges, digestive tract and urinary bladder. In: Penfield W (ed) Cytology and cellular pathology of the nervous system, vol 1. Hoeber, New York, pp 383–420Google Scholar
  72. Suzuki N, Hardebo JE, Owman C (1989) Origins and pathways of cerebrovascular nerves storing substance P and calcitonin gene-related peptide in rat. Neuroscience 31:427–438Google Scholar
  73. Tervo T, Joó F, Huikuri KT, Toth I, Palkama A (1979) Fine structure of sensory nerves in the rat cornea: an experimental nerve degeneration study. Pain 6:57–70Google Scholar
  74. Tsai S-H, Tew JM, McLean JH, Shipley MT (1988) Cerebral arterial innervation by nerve fibers containing calcitonin gene-related peptide (CGRP): I. Distribution and origin of CGRP perivascular innervation in the rat. J Comp Neurol 271:435–444Google Scholar
  75. Uddman R, Edvinsson L, Owman C, Sundler F (1981) Perivascular substance P: occurrence and distribution in mammalian pial vessels. J Cereb Blood Flow Metab 1:227–232Google Scholar
  76. Uddman R, Edvinsson L, Ekman R, Kingman T, McCulloch J (1985) Innervation of feline cerebral vasculature by nerve fibers containing calcitonin gene-related peptide: Trigeminal origin and co-existence with substance P. Neurosci Lett 62:131–136Google Scholar
  77. Uddman R, Hara H, Edvinsson L (1989) Neuronal pathways to the rat middle meningeal artery revealed by retrograde tracing and immunocytochemistry. J Auton Nerv Syst 26:69–75Google Scholar
  78. Wiesenfeld-Hallin Z, Hökfelt T, Lundley JM, Forssman 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
  79. Yeh Y, Kruger L (1984) Fine-structural characterization of the somatic innervation of the tympanic membrane in normal, sympathectomized, and neurotoxin-denervated rats. Somatosens Res 1:359–378Google Scholar
  80. Zagami AS, Goadsby PJ, Edvinsson L (1989) Extracranial peptide release following stimulation of the superior sagittal sinus in the cat. Cephalalgia 9 [Suppl 10]: 294–295Google Scholar

Copyright information

© Springer-Verlag 1993

Authors and Affiliations

  • Karl Meßlinger
    • 1
  • Ulrike Hanesch
    • 1
  • Matthias Baumgärtel
    • 1
  • Brigitte Trost
    • 1
  • Robert F. Schmidt
    • 1
  1. 1.Physiologisches Institut, Lehrstuhl II, Julius-Maximilians-Universität WürzburgWürzburgGermany

Personalised recommendations