Summary
Different techniques for the extraction and initial purification of porcine gastrointestinal glucagon-like immunoreactivity (GLI) were compared with reference to yield, and preservation of number and pattern of GLI components. The conventional acid-ethanol technique combined with ethanol-ether purification gave high yields and a reproducible pattern of components. Large amounts of tissue were more easily extracted using another technique, based on extraction by boiling, extraction and precipitation with acetone, and — if necessary — salting out. — By means of the latter two techniques mucosal tissue from all of the porcine gastrointestinal tract was extracted and subjected to gel filtration. Glucagon-like peptides were searched for using: — 1. a radioimmunoassay which quantifies gut type glucagon (GTG), as well as pancreatic type glucagon (PTG), 2. a radioimmunoassay highly specific for pancreatic type glucagon (PTG), and 3. a radioreceptor assay based on binding of glucagon to porcine liver cell membranes. — The oesophageal, the fundic, and the antro-pyloric parts of the gastric mucosa contained very small amounts of GLI. The cardiac gland region contained small amounts of a peptide indistinguishable from “true” glucagon. The duodenal mucosa contained small amounts of “true” glucagon and may be a smaller, glucagon-like peptide. The mucosa of the small intestine contained large amounts of both high and low molecular weight GTG and, in addition, PTG of high molecular weight and “true” glucagon. The colon also contained these components with “true” glucagon in high concentrations. Only small GTG and “true” glucagon were receptor-active, the former with less than its immunometric potency.
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Moody, A.J.: Gastrointestinal glucagon-like immunoreactivity. In: P.J. Lefebvre, R.H. Unger (Eds.): Glucagon, pp. 319–341. Oxford: Pergamon Press 1972
Assan, R.: Gut glucagon. In: S.A. Berson, R.S. Yalow (Eds.): Methods of investigative and diagnostic endocrinology, pp. 888–901, Vol. 2B, part III. Amsterdam, London: North-Holland Publ. Comp. 1973
Sasaki, H., Rubalcava, B., Baetens, D., Blasquez, E., Srikant, C.B., Orci, L., Unger, R.H.: Identification of glucagon in the gastrointestinal tract. J. Clin. Invest.56, 135–145 (1975)
Larsson, L.-I., Holst, J., H»kanson, R., Sundler, F.: Distribution and properties of glucagon immunoreactivity in the digestive tract of various mammals: An immuno-histochemical and immunochemical study. Histochemistry44, 281–290 (1975)
Sundler, F., Alumets, H., Holst, J., Larsson, L.-I., H»kanson, R.: Ultrastructural identification of cells storing pancreatic-type glucagon in dog stomach. Histochemistry50, 33–37 (1976)
Baetens, D., Rufener, C., Srikant, B.C., Dobbs, R., Unger, R., Orci, L.: Identification of glucagon producing cells (A cells) in dog gastric mucosa. J. Cell Biol.69, 455–464 (1976)
Bloom, S.R., Bryant, M.G., Polak, J.M.: Distribution of gut hormones. Gut16, 821 (1975)
Bustad, L.K., McClellan, R.O., Burns, M.P.: Swine in biomedical research. Proceedings of a Symposium at the Pacific Northwest Laboratory, Richland, Washington. Batelle Memorial Institute, 1966
Holst, J.J., Christiansen, J., Kühl, C.: The enteroglucagon response to intrajejunal infusion of glucose, triglycerides, and sodium chloride, and its relation to jejunal inhibition of gastric acid secretion in man. Scand. J. Gastroenterol.11, 297–304 (1976)
Heding, L.G.: Radioimmunological determination of pancreatic and gut glucagon in plasma. Diabetologia7, 10–19 (1971)
Albano, J.D.M., Ekins, R.P., Maritz, G., Turner, R.C.: A sensitive precise radioimmunoassay of serum insulin relying on charcoal separation of bound and free hormone moieties. Acta Endocrinol. (Kbh)70, 487–509 (1972)
Holst, J.J., Aasted, B.: Production and evaluation of glucagon antibodies for radioimmunoassay. Acta Endocrinol. (Kbh)77, 715–726 (1974)
Jørgensen, K.H., Larsen, U.D.: Purification of125I-glucagon by anion exchange chromatography. Horm. Metab. Res.4, 223–224 (1972)
Sundby, F., Jacobsen, H., Moody, A.J.: Purification and characterization of a protein from porcine gut with glucagon-like immunoreactivity. Horm. Metab. Res.8, 366–371 (1976)
Holst, J.J.: A radioreceptor-assay for glucagon: binding of enteroglucagon to liver plasma membranes. Diabetologia11, 211–219 (1975)
Best, C.H., Haist, R.E., Ridout, J.H.: Diet and insulin content of the pancreas. J. Physiol. (Lond)97, 107–115 (1939)
Mutt, V.: Preparation of highly purified secretin. Arkiv för Kemi (Uppsala)15, 69–95 (1959)
Kenny, A.J.: Extractable glucagon of the human pancreas. J. Clin. Endocrinol. Metab.15, 1089–1105 (1955)
Manns, J.G.: Separation of pancreatic and gut glucagonlike immunoreactivity (G.L.I.) with observations on plasma concentrations of these hormones during lactation. Can. J. Physiol. Pharmacol.50, 554–560 (1972)
Moody, A.J., Markussen, J., Schaich Fries, A., Steenstrup, C., Sundby, F., Malaisse, W., Malaisse-Lagae, F.: The insulin releasing activities of extracts of pork intestine. Diabetologia6, 135–140 (1970)
Markussen, J., Frandsen, E., Heding, L.G., Sundby, F.: Turkey glucagon: Crystallization, amino acid composition and immunology. Horm. Metab. Res.4, 360–363 (1972)
Sundby, F., Markussen, J.: Isolation, crystallization and amino acid composition of rat glucagon. Horm. Metab. Res.3, 184–187 (1970)
Sutherland, E.W., Cori, C.F., Haynes, R., Olsen, N.S.: Purification of the hyperglycemic-glycogenolytic factor from insulin and from gastric mucosa. J. Biol. Chem.180, 825–837 (1949)
Taylor, J.F.: The isolation of proteins. In: H. Neurath, K. Bailey (Eds.): The proteins, vol. I, part A, pp. 49–58. New York: Academic Press 1953
Sisson, S., Grossman, J.D.: The anatomy of the domestic animals, pp. 490–493. Philadelphia, London: W.B. Saunders Company 1953
Rigopoulou, D., Valverde, I., Marco, J., Faloona, G., Unger, R.H.: Large glucagon immunoreactivity in extracts of pancreas. J. Biol. Chem.245, 496–501 (1970)
Valverde, I., Rigopoulou, D., Exton, J., Ohneda, A., Eisentraut, A., Unger, R.H.: Demonstration and characterization of a second fraction of glucagon-like immunoreactivity in jejunal extracts. Am. J. Med. Sci.255, 415–419 (1968)
Murphy, R.F., Buchanan, K.D., Elmore, D.T.: Isolation of glucagon-like immunoreactivity of gut by affinity chromatography on anti-glucagon antibodies coupled to Sepharose 4B. Biochim. Biophys. Acta303, 118–127 (1973)
Bataille, D.P., Freychet, P., Kitabgi, P.E., Rosselin, G.E.: Gut glucagon: A common receptor site with pancreatic glucagon in liver cell plasma membranes. FEBS Lett.30, 215–218 (1973)
Valverde, I., Rigopoulou, D., Marco, J., Faloona, G., Unger, R.H.: Characterization of glucagon-like immunoreactivity (GLI). Diabetes19, 614–623 (1970)
Valverde, I., Rigopoulou, D., Marco, J., Faloona, G., Unger, R.H.: Molecular size of extractable glucagon and glucagon-like immunoreactivity (GLI) in plasma. Diabetes19, 624–629 (1970)
Langslow, D.R.: The action of gut glucagon-like immunoreactivity and other intestinal hormones on lipolysis in chicken adipocytes. Horm. Metab. Res.5, 428–432 (1973)
Bloom, S.R., Royston, C.M.S., Thompson, J.P.S.: Enteroglucagon release in the dumping syndrome. Lancet1972 II, 789–791
Dyck, W.P.: Influence of intrajejunal glucose on pancreatic secretion in man. Gastroenterology58, 1040 (1970)
Unger, R.H.: Diabetes and the alpha cell. Diabetes25, 136–151 (1976)
Unger, R.H., Orci, L.: The essential role of glucagon in the pathogenesis of diabetes mellitus. Lancet1975 I, 14–16
Dobbs, R., Sakurai, H., Sasaki, H., Faloona, G., Valverde, I., Baetens, D., Orci, L., Unger, R.H.: Glucagon: role in the hyperglycemia of diabetes mellitus. Science187, 544–547 (1975)
Sakurai, H., Dobbs, R.E., Unger, R.H.: The role of glucagon in the pathogenesis of the endogenous hyperglycemia of diabetes mellitus. Metabolism24, 1287–1297 (1975)
Matsuyama, T., Foa, P.P.: Plasma glucose, insulin, pancreatic, and enteroglucagon levels in normal and depancreatized dogs. Proc. Soc. Exp. Biol. Med.147, 97–102 (1974)
Vranic, M., Pek, S., Kawamori, R.: Increased “glucagon immunoreactivity” in plasma of totally depancreatized dogs. Diabetes23, 905–912 (1974)
Mashiter, K., Harding, P.E., Chou, M., Mashiter, G.D., Stout, J., Diamond, D., Field, J.B.: Persistent pancreatic glucagon but not insulin response to arginine in pancreatectomized dogs. Endocrinology96, 678–693 (1975)
Barnes, A.J., Bloom, S.R.: Pancreatectomized man: a model for diabetes without glucagon. Lancet1976 I, 219–221
Palmer, J.P., Werner, P.L., Benson, J.W., Ensinck, J.W.: Plasma glucagon after pancreatectomy. Lancet1976 I, 1290
Palmer, J., Benson, J., Werner, P., Ensinck, J.: Glucagon secretion in a totally pancreatectomized patient. Clin. Res.24, 119 A (1976)
Botha, J.L., Vinik, A.I.: Plasma glucagon after pancreatectomy. Lancet1976 I, 1290–1291
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Holst, J.J. Extraction, gel filtration pattern, and receptor binding of porcine gastrointestinal glucagon-like immunoreactivity. Diabetologia 13, 159–169 (1977). https://doi.org/10.1007/BF00745145
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DOI: https://doi.org/10.1007/BF00745145