Summary
Changes induced in the ultrastructure of the epithelium of the rat colon descendens by long-term electric field stimulation (EFS) in an Ussing chamber were investigated. The anion secretion, which was induced by EFS and was measured by the short-circuit current, fell continuously during a 5 h stimulation. At the end of the stimulation period, small particles were observed in the epithelium; these did not appear in unstimulated control tissue. They were localized predominantly in the apical part of the cell. By staining with periodic acidthiosemicarbazide-silver proteinate and because of their sensitivity to α-amylase, they were identified as glycogen deposits. This storage of glycogen was time-dependent and was first visible after an EFS of 2 h. It did not appear if glucose was substituted in the bathing solution by sodium butyrate. Glycogen particles were also observed after addition of forskolin, which in contrast to EFS causes a high secretory activity that is stable over several hours. The surface cells contained significantly more glycogen than the crypt cells when secretion was stimulated by EFS or forskolin. The formation of glycogen during EFS was not prevented by tetrodotoxin (TTX). In contrast, TTX itself, which causes maximal absorptive activity by blocking secretomotor neurons, induced the appearance of glycogen in the enterocytes without EFS. However, in the presence of TTX, the amount of glycogen was the same in surface and crypt cells. The results demonstrate that the capacity to synthesize and store glycogen, which has up to now only been observed in embryonic or tumor epithelial cells, is still present in adult colonic mucosa. Procedures carried out to change the functional state of the epithelium seem to induce, at least in vitro, a disinhibition of this capacity.
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References
Andres H, Bock R, Bridges RJ, Rummel W, Schreiner J (1985) Submucosal plexus and electrolyte transport across rat colonic mucosa. J Physiol 364:301–312
Binder HJ, Sandle GI (1987) Electrolyte absorption and secretion in the mammalian colon. In: Johnson LR (ed) Physiology of the gastrointestinal tract. Raven Press, New York, pp 1389–1418
Bridges RJ, Rummel W, Simon B (1983) Forskolin induces chloride secretion across the isolated mucosa of rat colon descendens. Naunyn-Schmiedeberg's Arch Pharmacol 323:355–360
Carey HV, Cooke HJ, Nemeth PR, Zafirov DH, Wood JD (1985a) Nerve-mediated action of forskolin on guinea-pig ileal mucosa. Experientia 41:1156–1158
Carey HV, Cooke HJ, Zafirova M (1985b) Mucosal responses evoked by stimulation of ganglion cell somas in the submucosal plexus of the guinea-pig ileum. J Physiol 364:69–79
Catterall WA (1980) Neurotoxins that act on voltage-sensitive sodium channels in excitable membranes. Ann Rev Pharmacol Toxicol 20:15–43
Coimbra A (1966) Evaluation of the glycogenolytic effect of alpha-amylase using autoradioraphy and electron microscopy. J Histochem 154:898–906
Cooke HJ (1986) Neurobiology of the intestinal mucosa. Gastroenterology 90:1057–1081
DeHaan RL, Ursprung H (1965) Organogenesis. Holt, Rinehart and Winston. New York Chicago San Francisco Toronto London
Diener M, Bridges RJ, Hubel KA, Rummel W (1987) The effect of electric field stimulation on ion transport in rat colon descendens Z Gastroenterologie 25:383
Diener M, Knobloch SF, Rummel W (1988) Action of loperamide on neuronally mediated and Ca2+-or cAMP-mediated secretion in rat colon. Eur J Pharmacol 152:217–225
Diener M, Mestres P, Bridges RJ, Rummel W (1989a) Functional and morphological changes during electric field stimulation of rat colon descendens. In: Singer MV, Goebell H (eds) Nerves and the gastrointestinal tract. MTP Press, Lancaster, pp 705–712
Diener M, Rummel W, Mestres P, Lindemann B (1989b) Single chloride channels in colon mucosa and isolated colonic enterocytes of the rat. J Membrane Biol 108:21–30
Friden J, Seger J, Ekblom B (1985) Implementation of periodic acid-thiosemicarbazide-silver proteinate staining for ultrastructural assessment of muscle glycogen utilization during exercise. Cell Tissue Res 242:229–232
Hoefsloot LH, Hoogeveen-Westerveld M, Kross MA, Van Beeumen J, Reuser AJ, Üostra BA (1988) Primary structure and processing of lysosomal alpha-glucosidase; homology with the intestinal sucrase-isomaltase complex. EMBO J 7:1697–1704
Hubel KA (1978) The effects of electrical field stimulation and tetrodotoxin on ion transport by the isolated rabbit ileum. J Clin Invest 62:1039–1047
Mosher HS, Fuhrmann FA, Buchwald HD, Fischer HG (1964) Tarichotoxin-tetrodotoxin: a potent neurotoxin. Science 144:1100–1110
Nemeth PR, Zafirov D, Wood JD (1985) Forskolin mimics slow synaptic excitation in myenteric neurons. Eur J Pharmacol 101:303–304
Nemeth PR, Palmer JM, Wood JD, Zafirov DH (1986) Effect of forsko in on electrical behaviour of myenteric neurones in guinea-pig small intestine. J Physiol 376:439–450
Parsons DS, Paterson CR (1965) Fluid and solute transport across rat colonic mucosa. Quart J Exp Physiol 50:220–231
Revel JP (1964) Electron microscopy of glycogen. J Histochem Cytochem 12:104–114
Richardson KC, Jarrett L, Finke H (1960) Embedding in epoxy resins for ultrathin sectioning in electron microscopy. Stain Technol 35:313–323
Roediger WEW (1982) Utilization of nutrients by isolated epithelial cells of the rat colon. Gastroenterology 83:424–429
Rousset M, Dussaulx E, Chevalier G, Zweibaum A (1980) Growthrelated glycogen levels of human intestine carcinoma cell lines grown in vitro and in vivo in nude mice. J Natl Cancer Res 65:885–889
Rousset M, Laburthe M, Chevalier G, Boissard C, Rosselin G, Zweibaum A (1981) Vasoactive intestinal peptide (VIP) control of glycogenolysis in the human colon carcinoma cell line HT-29 in culture. FEBS Lett 126:38–40
Seamon KB, Padgett W, Daly JW (1981) Forskolin: unique diterpene activator of adenylate cyclase in membrane and in intact cells. Proc Natl Acad Sci USA 78:3363–3367
Smith RE, Farquhar MG (1966) Lysosome function in the regulation of the secretory process of the anterior pituitary gland. J Cell Biol 31:319–347
Thiéry JP (1967) Mise en évidence des polysaccharides sur coupes fines en microscopie électronique. J Microsc 6:987–1018
Vollrath L (1969) Über die Entwicklung des Dünndarms der Ratte. Morphologische, histochemische und experimentelle Untersuchungen. Erg Anat Entw Gesch 41:3–66
Wattiaux R (1969) Biochemistry and function of lysosomes. In: Lima-De-Faria A (ed) Handbook of molecular cytology. North-Holland, Amsterdam London, pp 1159–1178
Welsh MJ, Smith PL, Fromm M, Frizzell RA (1982) Crypts are the site of intestinal fluid and electrolyte secretion. Science 218:1219–1221
Wille KH (1988) Über das Glykogen im fetalen Darmepithel. Anat Anz 167:129–139
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Mestres, P., Diener, M. & Rummel, W. Storage of glycogen in rat colonic epithelium during induction of secretion and absorption in vitro. Cell Tissue Res 261, 195–203 (1990). https://doi.org/10.1007/BF00329452
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DOI: https://doi.org/10.1007/BF00329452