Abstract
In gastrointestinal muscles special cells, referred to as interstitial cells, may be involved in pacemaking and transduction of inputs from the enteric nervous system. We have used a modification of the NADH diaphorase method to characterize the distribution of interstitial cells in the muscularis externa of the canine colon. The staining product of the NADH diaphorase reaction is useful because it allows light and electron microscopic studies to be performed with the same marker. Therefore rigorous identification of the cells observed at the light microscopic level could be made by electron microscopy. We were able to lable at least three classes of interstitial cells: (1) at the submucosal surface of the circular muscle layer; (2) within the thickness of the circular and longitudinal muscle layers; and (3) in the region of the myenteric plexus. This technique also labeled cell bodies and initial segments of processes of Dogiel type II neurones in enteric ganglia. Nerve fibres within the muscle layers did not exhibit NADH diaphorase activity. This study has identified the interstitial cells within the circular and longitudinal muscle layers and shows the arrangement of these cells in a three-dimensional network.
Similar content being viewed by others
References
Berezin I, Huizinga JD, Daniel EE (1988) Interstitial cells of Cajal in the canine colon: a special communication network at the inner border of the circular muscle. J Comp Neurol 273:42–51
Berezin I, Huizinga JD, Daniel EE (1990) Structural characteristics of interstitial cells of Cajal in myenteric plexus and muscle layers of canine colon. Can J Physiol Pharmacol 68:1419–1431
Bredt DS, Snyder SH (1990) Isolation of nitric oxide synthase, a calmodulin-requiring enzyme. Proc Natl Acad Sci USA 87:682–685
Christensen JA (1992) Commentary on the morphological identification of interstitial cells of Cajal in the gut. J Auton Nerv Syst 37:75–88
Christensen J, Rick GA (1987) Intrinsic nerves in the mammalian colon: confirmation of a plexus at the circular muscle-submucosal interface. J Auton Nerv Syst 21:223–231
Costa M, Furness JB (1973) The simultaneous demonstration of adrenergic fibres and enteric ganglion cells. Histochem J 5:343–349
Costa M, Furness JB, Llewellyn-Smith IJ (1987) Histochemistry of the enteric nervous system. In: Johnson LR (ed) Physiology of the gastrointestinal tract, 2nd edn. Raven Press, New York, pp 1–40
Csoknya M, Fekete E, Gabriel R, Halasy K, Benedeczky I (1990) Histochemical characterization of myenteric plexus in domestic fowl small intestine. Z Mikrosk Anat Fosch 104:625–638
Daniel EE, Berezin I (1992) Interstitial cells of Cajal: are they major players in control of gastrointestinal motility? J Gastrointest Motil 4:1–24
Dawson TM, Bredt DS, Fotuhi M, Hwang PM, Snyder SH (1991) Nitric oxide synthase and neuronal NADPH diaphorase are identical in brain and peripheral tissues. Proc Natl Acad Sci USA 88:7797–7801
Durdle NG, Kingman YJ, Bowes KK, Chambers MM (1983) Origin of slow waves in the canine colon. Gastroenterology 84:375–382
Edwards YH, Potter J, Hopkinson DA (1980) Human FAD-dependent NAD(P)H diaphorase. Biochem J 187:429–436
Faussone-Pellegrini MS (1987) Cytodifferentiation of the interstitial cells of Cajal of mouse colonic circular muscle layer. Acta Anat 128:98–109
Faussone-Pellegrini MS, Cortesini C, Pantalone D (1990a) Neuromuscular structures specific to the submucosal border of the human colonic circular muscle layer. Can J Physiol Pharmacol 68:1437–1446
Faussone-Pellegrini MS, Pantalone D, Cortesini C (1990b) Smooth muscle cells, interstitial cells of Cajal and myenteric plexus interrelationships in the human colon. Acta Anat 139:31–44
Furness JB, Bornstein JC, Trussell DC (1988) Shapes of nerve cells in the myenteric plexus of the guinea-pig small intestine revealed by the intracellular injection of dye. Cell Tissue Res 254:561–571
Gabella G (1969) Detection of nerve cells by a histochemical technique. Experientia 25:218–219
Gabella G (1990) On the plasticity of form and structure of enteric ganglia. J Auton Nerv Syst 30:S59-S66
Gabriel R, Benedeczky I, Csoknya M (1989) Myenteric plexus of frog large intestine: light and electron microscopy of fiber system and neurons. Acta Morphol Hung 37:71–84
Gabriel R, Halasy K, Csoknya M (1988) Visualization of neurons by NADH-diaphorase staining in the myenteric plexus of some invertebrate and vertebrate species. Z Mikrosk Anat Forsch 102:769–784
Hope BT, Michael GJ, Knigge KM, Vincent SR (1991) Neuronal NADPH diaphorase is a nitric oxide synthase. Proc Natl Acad Sci USA 88:2811–2814
Hoyle CH, Burnstock G (1989) Neuronal populations in the submucous plexus of the human colon. J Anat 166:7–22
Komuro T (1982) The interstitial cells in the colon of the rabbit. Cell Tissue Res 222:41–51
Langton P, Ward SM, Carl A, Norell MA, Sanders KM (1989) Spontaneous electrical activity of interstitial cells of Cajal isolated from canine proximal colon. Proc Natl Acad Sci USA 86:7280–7284
Lee HK, Sanders KM (1993) Comparison of ionic currents from interstitial cells and smooth muscle cells of canine colon. J Physiol 460:135–152
Liu LWC, Daniel EE, Huizinga JD (1992) Excitability of canine colon circular muscle disconnected from the network of interstitial cells of Cajal. Can J Physiol Pharmacol 70:289–295
Liu LWC, Thuneberg L, Daniel EE, Huizinga JD (1993) Selective accumulation of methylene blue by interstitial cells of Cajal in canine colon. Am J Physiol 264:G64-G73
Mitchell JA, Kohlhaas KL, Matsumoto T, Pollock JS, Forstermann U, Warner TD, Schmidt HHW, Murad F (1992) Induction of NADPH-dependent diaphorase and nitric oxide synthase activity in aortic smooth muscle and cultured macrophages. Mol Pharmacol 41:1163–1168
Pompolo S, Furness JB (1988) Ultrastructure and synaptic relationships of calbindin-reactive, Dogiel type II neurons, in myenteric ganglia of guinea-pig small intestine. J Neurocytol 17:771–782
Schultzberg M, Segura-Aguilar J, Lind C (1988) Distribution of DT diaphorase in the rat brain: biochemical and immunohistochemical studies. Neuroscience 27:763–776
Serio R, Barajas-Lopez C, Daniel EE, Berezin I, Huizinga JD (1991) Slow wave activity in colon: role of network of submucosal interstitial cells of Cajal. Am J Physiol 260:G636-G645
Smith TK, Reed JB, Sanders KM (1987) Origin and propagation of electrical slow waves in circular muscle of canine proximal colon. Am J Physiol 252:215–224
Thuneberg L (1982) Interstitial cells of Cajal: intestinal pacemaker cells? Adv Anat Embryol Cell Biol 71:1–130
Thuneberg L (1982) Interstitial cells of Cajal. In: Wood JD (ed) Handbook of physiology: the gastrointestinal system, vol 1, pt 1. American Physiological Society, Bethesda, Maryland, pp 349–386. Distributed by Oxford University Press, New York
Vincent SR, Johansson O (1983) Striatal neurons containing both somatostatin-and avian pancreatic polypeptide (APP)-like immunoreactivities and NADPH-diaphorase activity: a light and electron microscopic study. J Comp Neurol 217:264–270
Ward SM, Sanders KM (1990) Pacemaker activity in septal structures of canine colonic circular muscle. Am J Physiol 259:G264-G273
Ward SM, Keller R, Sanders KM (1991) Structure and organization of the electrical activity of the canine distal colon. Am J Physiol 260:G724-G735
Ward SM, Xue C, Shuttleworth CW, Bredt DS, Snyder SH, Sanders KM (1992) NADPH diaphorase and nitric oxide synthase co-localization in enteric neurons of the canine proximal colon. Am J Physiol 263:G277-G284
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Xue, C., Ward, S.M., Shuttleworth, C.W. et al. Identification of interstitial cells in canine proximal colon using NADH diaphorase histochemistry. Histochemistry 99, 373–384 (1993). https://doi.org/10.1007/BF00717050
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00717050