Abstract
Nitric oxide has been implicated in mechanisms mediating nerve-evoked vasodilatory and secretory responses in salivary glands. In the present study, the occurrence and distribution of nitric oxide synthase (NOS)-immunoreactive nerves in ferret and rat salivary glands were investigated using immunocytochemistry with rabbit and sheep NOS antisera, and using NADPH-diaphorase enzyme histochemistry. In the parotid, submandibular and sublingual glands of the rat and the ferret, NOS-immunoreactive varicose terminals encircled acini and arteries of various sizes. In the ferret, collecting ducts were also supplied with NOS-immunoreactive fibres. In the rat, only the granular ducts of the submandibular gland were supplied with such fibres. The NOS-immunoreactive innervation of acinar cells was more abundant in the rat than in the ferret, whereas the opposite was true for the innervation of blood vessels. No NOS immunoreactivity was observed in the vascular endothelium. In both species, NOS-positive ganglionic cell bodies were found in the hilar regions of the submandibular and sublingual glands, whereas none could be detected in the parotid glands. NADPH-diaphorase reactivity had the same neuronal distribution as NOS immunoreactivity and, in addition, NADPH-diaphorase reactivity was expressed in ductal epithelium. Neither sympathetic denervation (by removal of the superior cervical ganglion) nor treatment with the sensory neurotoxin capsaicin reduced the NOS-immunoreactive innervation of the parotid gland. However, parasympathetic denervation (by cutting the auriculo-temporal nerve) caused an almost total disappearance of the NOS-immunoreactive innervation. The present findings provide a morphological background to the suggested role of nitric oxide in parasympathetic secretory and vascular responses of salivary glands.
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References
Alm, P. & EkstrÖm, J. (1976) Cholinergic nerves of unknown origin in the parotid glands of rats. Arch. Oral. Biol. 21, 417-21.
Alm, P., Larsson, B., Ekblad, E., Sundler, F. & Andersson, K.-E. (1993) Immunohistochemical localization of peripheral nitric oxide synthase-containing nerves using antibodies raised against synthesized C-and N-terminal fragments of a cloned enzyme from rat brain. Acta Physiol. Scand. 148, 421-9.
Alm, P., Uvelius, B., EkstrÖm, J. Holmquist, B., Larsson, B. & Andersson, K.-E. (1995) Nitric oxide synthase-containing neurons in rat parasympathetic, sympathetic and sensory ganglia: a comparative study. Histochem. J. 27, 819-31.
AsztÉly, A., Tobin, G. & EkstrÖm, J. (1994a) Masticatory-salivary reflexes mobilize non-adrenergic, non-cholinergic secretory mechanisms in parotid glands of rats. Acta Physiol. Scand. 151, 373-6.
AsztÉly, A., Tobin, G. & EkstrÖm, J. (1994b) Parasympathetic non-adrenergic, non-cholinergic reflex secretion of parotid acinar granules in rats pretreated with atropine and adrenoceptor antagonists. Reg. Pept. 51, 255-62.
Bloom, S.R. & Edwards, A.V. (1980) Vasoactive intestinal peptide in relation to atropine resistant vasodilatation in the submaxillary gland of the cat. J. Physiol. 300, 41-53.
Blottner, D., Grozdanovic, Z. & Gossrau, R. (1995) Histochemistry of nitric oxide in the nervous system. Histochem. J. 27, 785-811.
Bredt, D.S. (1995) Molecular characterization of nitric oxide synthase. In Nitric Oxide Synthase in the Nervous System (edited by Vincent, S.), pp. 1-19. London: Academic Press, Harcourt Brace & Company.
Buckle, A.D., Parker, S.J., Bloom, S.R. & Edwards, A.V. (1995) The role of nitric oxide in the control of protein secretion in the submandibular gland of the cat. Exp. Physiol. 80, 1019-30.
Ceccatelli, S., Lundberg, J.M., Zhang, X., Åman, K. & HÖkfelt, T. (1994) Immunohistochemical demonstration of nitric oxide synthase in the peripheral autonomic nervous system. Brain Res. 665, 381-95.
Dail, G.W., Barba, V., Leyba, L & Galindo, R. (1995) Neural and endothelial nitric oxide synthase activity in rat penile erectile tissue. Cell Tiss Res. 282, 109-16.
Dawson, T.M., Bredt, D.S., Fotuhi, M., Hwang, P.M. & Snyder, S.S. (1991) Nitric oxide synthase and neuronal NADPH diaphorase are identical in brain and peripheral tissues. Proc. Natl Acad. Sci. USA 88, 7797-801
Edwards, A.V. & Garrett, J.R. (1993) Nitric oxiderelated vasodilator responses to parasympathetic stimulation of the submandibular gland in the cat. J. Physiol. 464, 379-92.
Edwards, A.V., Tobin, G., EkstrÖm, J, & Bloom, S.R. (1996). The role of nitric oxide during release of the peptide VIP from parasympathetic nerve terminals in the submandibular gland of the anesthetized cat. Exp. Physiol. 81, 349-59.
EkstrÖm, J. (1974) Choline acetyltransferase activity in rat salivary glands after cellulose rich diet or treatment with an atropine-like drug. Q. J. Exp. Physiol. 59, 191-9.
EkstrÖm, J. (1987) Neuropeptides and Secretion. J. Dent. Res. 66(2), 524-30.
EkstrÖm, J. (1989) Autonomic control of the salivary secretion. Proc. Finn. Dent. Soc. 85(4/5), 323-31.
EkstrÖm, J. & Olgart, L. (1986) Complementary action of substance P and vasoactive intestinal peptide on the rat parotid secretion. Acta Physiol. Scand. 126, 25-31.
EkstrÖm, J. & Tobin, G. (1989) Secretion of protein from salivary glands in the ferret in response to vasoactive intestinal peptide. J Physiol. 415, 131-41.
EkstrÖm, J., MÅnsson, B., Tobin, G., Garret, J.R. & Thulin, A. (1983) Atropine-resistant secretion of parotid saliva on stimulation of the auriculo-temporal nerve in the rat. Acta Physiol. Scand. 119, 445-9.
EkstrÖm, J., Brodin, E., HÅkanson, R. & Sundler, F. (1984) Vasoactive intestinal polypeptide and substance P in salivary glands of the rat following denervation or duct ligation. Reg. Pept. 10, 1-10.
EkstrÖm, J., Brodin, E., Ekman, R. HÅkanson, R., MÅnsson, B. & Tobin, G. (1985) Depletion of neuropeptides in rat parotid glands and declining atropine-resistant salivary secretion upon continuous parasympathetic nerve stimulation. Reg. Pept. 11, 353-9.
EkstrÖm, J., MÅnsson, B. & Tobin, G. (1987) Non-adrenergic, non-cholinergic parasympathetic secretion in the rat submaxillary and sublingual glands. Pharmacol. Toxicol. 60, 284-7.
EkstrÖm, J., Olgart, L., MÅnsson, B. & Tobin, G. (1988a) Non-adrenergic, non-cholinergic salivary secretion in the ferret. Q. J. Exp. Physiol. 73, 163-73.
EkstrÖm, J., Ekman, R., HÅkanson, R., SjÖgren, S. & Sundler, F. (1988b) Calcitonin gene-related peptide in rat salivary glands: neuronal localization, depletion upon nerve stimulation, and effects on salivation in relation to substance P. Neuroscience 26, 933-49.
EkstrÖm, J., Helander, H.F. & Tobin, G. (1993) Parasympathetic non-adrenergic non-cholinergic mechanisms in reflex secretion of parotid acinar granules in conscious rats. J. Physiol. 472, 233-44.
EkstrÖm, J., Ekman, R. Luts, A., Sundler, F. & Tobin, G. (1996) Neuropeptide Y in salivary glands of the rat: origin, release and secretory effects. Reg. Pept. 61, 125-34.
ErÄnkÖ, O. & HÄrkÖnen, M. (1964) Noradrenaline and acetylcholinesterase in sympathetic ganglion cells of the rat. Acta Physiol. Scand. 61, 299-300.
Grozdanovic, Z., Baumgarten, H.G. & BrÜning, G. (1992) Histochemistry of NADPH-diaphorase, a marker for neural nitric oxide synthase, in the peripheral autonomic nervous system of the mouse. Neuroscience 48, 225-35.
Heidenhain, R. (1972) Über die Wirkung einiger Gifte auf die Nerven der Glandula submaxillaris. Arch. Ges. Physiol. 5, 309-18.
Herbison, A.E., Simonian, S.X., Norris, P.J. & Emson, P.C. (1996) Relationship of neuronal nitric oxide synthase immunoreactivity to GnRH neurons in the ovariectomized and intact female rat. J. Neuroendocrinol. 8, 73-82.
Junqueria, L., Fajer, A., Rabinovitch, M. & Frankenthal, L. (1949) Biochemical and histochemical observations on the sexual dimorphism of mice submaxillary glands. J. Cell Comp. Physiol. 34, 129-58.
Koelle, G.B. (1955) The histochemical identification of acetylcholinesterase in cholinergic, adrenergic and sensory neurons. J. Pharmacol. Exp. Ther. 114, 167-84.
Lohinai, Z.S., SzÉkely, A.D., SoÓs, L. & FehÉr, E. (1995) Distribution of nitric oxide synthase containing elements in the feline submandibular gland. Neurosci. Lett. 192, 9-12.
Lundberg, J.M., Martling, C.-R. & HÖkfelt, T. (1988a) Airways, oral cavity and salivary glands: classical transmitters and peptides in sensory and autonomic neurons. In Handbook of Chemical neuroanatomy. vol. 6: The Peripheral Nervous System. (edited by BjÖrklund, A., HÖkfelt, T. & Owman, C.), pp. 391-444. Amsterdam: Elsevier Science Publishers B.V.
Lundberg, J.M., ÄnggÅrd, A., Fahrenkrug, J., HÖkfelt, T. & Mutt, V. (1980) Vasoactive intestinal polypeptide in cholinergic neurons of exocrine glands: Functional significance of coexisting transmitters for vasodilatation and secretion. Proc. Natl Acad. Sci. USA 77, 1651-5.
Lundberg, L.-M., Alm, P., Wharton, J. & Polak, J.M. (1988b) Protein gene product 9.5 (PGP 9.5) A new neuronal marker visualizing the whole uterine innervation and pregnancy-induced and developmental changes in the guinea pig. Histochemistry 90, 9-17.
Modin, A., Weitzberg, E., HÖkfelt, T. & Lundberg, J.M. (1994a) Nitric oxide synthase in the pig autonomic nervous system in relation to the influence of NG-nitro-L-arginine on sympathetic and parasympathetic control in vivo. Neuroscience 62, 189-203.
Modin, A., Weitzberg, E. & Lundberg, J.M. (1994b) Nitric oxide regulates peptide release from parasympathetic nerves and vascular reactivity to vasoactive intestinal polypeptide in vivo. Eur. J. Pharmacol. 261, 185-197.
Moncada, S. (1992) The 1991 Ulf von Euler Lecture. The L-arginine: nitric oxide pathway. Acta Physiol. Scand. 145, 201-27.
MÅnsson, B. & EkstrÖm, J. (1991) On the non-adrenergic, non-cholinergic contribution to the parasympathetic nerve-evoked secretion of parotid saliva in the rat. Acta Physiol. Scand. 141, 197-205.
Pinkstaff, C.A. (1972) Sexual dimorphism of the miniature pig submandibular glands. Am. J. Anat. 135, 371-79.
Schultz, T., Soinila, J., Tolonen, R., HÄppÖlÄ, O., Uusitalo, H. & Salo, A. (1994) The sympathetic and parasympathetic nature of neuropeptide Y-immunoreactive nerve fibres in the major salivary glands of the rat. Histochem. J. 26, 563-70.
Sharkey, K.A., Mathison, R., Sharif, M.N. & Davison, J.S. (1989) The effects of streptozotocin-induced diabetes on the peptidergic innervation and function of the rat parotid gland. J. Auton. Nerv. Syst. 27, 127-38.
Thulin, A. (1976) Blood flow changes in the submaxillary gland of the rat on parasympathetic and sympathetic nerve stimulation. Acta Physiol. Scand. 97, 104-9.
Tobin, G., Luts, A., Sundler, F. & EkstrÖm, J. (1990) VIP-containing nerve fibres in the submandibular gland of the dog and protein secretion in vitro in response to VIP. Reg. Pept. 29, 1173-77.
Tobin, G., EkstrÖm, J., Bloom, S.R. & Edwards, A.V. (1991) Atropine resistant submandibular responses to stimulation of the parasympathetic innervation in the anaesthetized ferret. J. Physiol. 437, 327-39.
Tobin, G., AsztÉly, A., Edwards, A.V., EkstrÖm, J., HÅkanson, R. & Sundler, F. (1995) Presence and effects of pituitary adenylate cyclase activating peptide (PACAP) in the submandibular gland of the ferret. Neuroscience 66, 227-35.
Tobin, G., Edwards, A.V., Bloom, S.R. & EkstrÖm, J. (1997) Nitric oxide in the control of submandibular function in the anaesthetized ferret. Exp. Physiol., 82, in press.
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Alm, P., Ekstrzm, J., Larsson, B. et al. Nitric oxide synthase immunoreactive nerves in rat and ferret salivary glands, and effects of denervation. J Mol Hist 29, 669–676 (1997). https://doi.org/10.1023/A:1026452715555
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DOI: https://doi.org/10.1023/A:1026452715555