The immune-ultrastructural investigations on adrenomedullin (AM) distribution in the pancreas of amphibian Rana esculenta

The immune-ultrastructural investigations on adrenomedullin (AM) distribution in the pancreas of amphibian Rana esculenta

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Title: The immune-ultrastructural investigations on adrenomedullin (AM) distribution in the pancreas of amphibian Rana esculenta
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Article_Title: The immune-ultrastructural investigations on adrenomedullin (AM) distribution in the pancreas of amphibian Rana esculenta
Authors: Ioana Trandaburu, Wolfgang Kummer, Tiberiu Trandaburu
Affiliation: Center of Cytobiology, Institute of Biology, Splaiul Independenţei 296, 060031 Bucharest, Romania
Institute of Anatomy and Cell Biology, Justus Liebig University, Aulweg 123, 35392 Giessen, Germany
Laboratory of Histology and Embryology, Faculty of Sciences, University of Piteşti, Târgul din Vale 1, 110040 Romania
Abstract: The pancreas of the frog Rana esculenta was investigated in electron microscopy for the immunocytochemical detection of the adrenomedullin (AM). These investigations revealed that AM didn’t label the secretory granules matrix of all endocrine cells, but rather several minority subpopulations of AM, SOM, PP and INS cells. Concretely, the results indicated that 10%-15% from glucagon (GLUC) producing cells, 4%-8% from somatostatin (SOM) and pancreatic polypeptide (PP) cells and only 2%-5% from insulin (INS) cells contained also AM. The sites of antigen-antibody reaction, labelled with colloidal gold particles, have been detected in the cells cytoplasm, randomly distributed among the secretory granules, surrounding them, or overlapped on the hormonal cores. In the exocrine pancreas, reactive nerve fibbers to AM occurred distributed among acini, in the ducts walls, or in those of blood vessels. The above findings and their functional significances are discussed in connection with the results previously reported in mammals and other lower vertebrates.
Keywords: adrenomedullin, AM, frog, pancreas, immunocytochemistry
References: Trandaburu I, Olaru L, Trandaburu T. The occurrence and distribution of adrenomedullin immunoreactivity in the gut of amphibians and reptiles. Proc Inst Biol. 2000; 3: 471-78.
Trandaburu I., Trandaburu T. The occurrence and distribution of adrenomedullin (AM) in the endocrine pancreas of several poikilotherm vertebrates: an immunohistochemical study. Studia Universitatis “Vasile Goldis” Life Sciences. 2010; 20: 27-33.
Kitamura K, Kanagawa K, Kawamoto M, Ichiki Y, Nakamura S, Matsuo H, Eto T. Adrenomedullin: a novel hypotensive peptide isolated from human pheochromocytoma. Biochem Biophys Res Commun. 1993; 192: 533-60.
Wimalawansa SJ. Amylin, calcitonin gene-related peptide, calcitonin, and adrenomedullin: a peptide superfamily. Crit Rev Neurobiol. 1977; 11: 167-239.
Olaru L. Contrbutions regarding the cytoarhitecture and immunohistochemistry of the diffuse neuroendocrine gastro-entero-pancreatic (GEP) system in amphibians. (doctoral thesis), 2005.
Ishiyama Y, Kitamura K, Ichiki Y, Nakamura S, Kida O, Kanagawa K, Eto T. Hemodynamic effects of a novel hypotensive peptide, human adrenomedullin, in rats. Eur J Pharmacol. 1933; 241: 271-73.
Nuki C, Kawasaki H, Kitamura K, Takenaga M, Kanagawa K, Eto T, Wada A. Vasodilator effect of adrenomedullin and calcitonin gene-related peptide receptors in rat mesenteric vascular beds. Biochem Biophys Res Commun. 1993; 196: 245-51.
Allen MA, Ferguson AY. In vitro recordings from area postrema neurons demonstrate responsiveness to adrenomedullin. Am J Physiol. 1996; R 920-R 925.
Kanazawa H, Kurihara N, Hirota K, Kudoh S, Kawaguchi T, Takeda T. Adrenomedullin, a novel discovered hypotensive peptide, is a potent bronchodilator. Biochem Biophys Res Commun. 1994; 205: 251-54.
Miller MJ, Martinez A, Unsworth E, Thiele CJ, Moody TW, Cuttitta F. Adrenomedullin: expression in human tumor cell lines and its potential role as an autocrine growth factor. J Biol Chem. 1996; 271:23345-351.
Walsh TJ, Martinez A, Peter J, Unsworth EJ, Cuttitta F. Antimicrobial activity of adrenomedullin and its gene-related peptide. J Biol Chem. 1998; 273: 3421-428.
Kato F, Kitamura K, Niina H, Yomamoto R, Washimine H, Kanagawa K, Yamamoto Y, Kobayashi H, Eto T, Wada A. Proadrenomedullin N-terminal 20 peptide (PAMP), an endogenous anticholinergic peptide: its exocytotic secretion and inhibition of catecholamine secretion in adrenal medulla. J Neurochem. 1995; 64: 459-61.
Mazzocchi G, Musajo F, Neri G, Gottardo G, Nussdorfer G. Adrenomedullin stimulates steroid secretion by the isolated perfused rat adrenal gland in situ: comparison with calcitonin gene-related peptide effects. Peptides. 1996; 17: 852-57.
Samson WK, Murphy T, Schell DA. A novel vasoactive peptide, adrenomedullin, inhibits pituitary adrenocorticotropin release. Endocrinology. 1995; 136: 2349-352.
Mulder H, Ahrén B, Karlsson S, Sundler F. Adrenomedullin: localization in the gastrointestinal tract and effect on insulin secretion. Regul Pept. 1996; 62: 107-12.
Ishizaka Y, Ishizaka Y, Tanaka M, Kitamura K, Kanagawa K, Minamino N, Matsuo H, Eto T. Adrenomedullin stimulates cyclic AMP formation in rat vascular smooth muscle cells. Biochem Biophys Res Commun. 1994; 200: 642-46.
Washimine H, Asada Y, Kitamura K, Ichiki Y, Hara S, Yamamoto Y, Kanagawa K, Sumiyoshi A, Eto T. Immunohistochemical identification of adrenomedullin in human, rat, and porcine tissues. Histochemistry. 1995; 103: 251-54.
Sakata J, Asada Y, Shimokubo T, Kitani H, Inatsu H, Kitamura K, Kanagawa K, Matsuo H, Sumiyoshi A, Eto T. Adrenomedullin in the gastrointestinal tract. Distribution and gene expression in rat and augmented gastric adrenomedullin after fasting. J Gastroenterol. 1998; 33: 828-34.
Asada Y, Seiichiro H, Marutsuka K, Kitamura K, Fuji T, Sakata J, Sato J, Kisanuki A, Eto T, Sumiyoshi A. A novel distribution of adrenomedullin immunoreactive cells in human tissues. Cell Tiss Res. 1999; 292: 45-58.
Lopez J, Cuesta N. Adrenomedullin as a pancreatic hormone.Microsc Res Techn. 2002; 57: 61-75.
Marutsuka K, Hatekayama K, Sato Y, Yamashita A, Sumiyoshi A, Asada Y. Immunohistological localization and possible functions of adrenomedullin. Hypertens Res. 2003; 26: 33-40.
Satoh F, Takahashi K, Murakami O, Totsune K, Sone M, Ohneda M, Abe K, Miura J, Hayashi Y, Sasano H, Mouri T. Adrenomedullin in human brain, adrenal glands and tumor tissues of pheochromocytoma, ganglioneuroblastoma and neuroblastoma. J Clin Endocrinol Metabol. 1995; 80: 1750-752.
Pio R, Martinez A, Cuttitta F. Cancer and diabetes: two pathological conditions in which adrenomedullin may be involved. Peptides. 2001; 22: 1719-729.
Gonzales A, Marin O, Sanchez-Camacho C, Pena JJ, Zudaire JJ, Martinez A, Cuttitta F, Munoz M. Localization of adrenomedullin-like immunoreactivity in the hypothalamo-hypophysial system of amphibians. Neurosci Lett. 1998; 242:13-16.
Lopez J, Cuesta N, Cuttitta F, Martinez A. Adrenomedullin in nonmammalian vertebrate pancreas: an immunocytochemical study. Gen Comp Endocrinol. 1999; 3: 309-22.
Zamboni L, De Martino C. Buffered picric acidformaldehyde: a new rapid fixative for electron microscopy. J Cell Biol. 1967; 35: 148A.
Carlemalm E, Garavito RM, Willinger W. Resin development for electron microscopy and an analysis of embedding at low temperature. J Microsc. 1982; 126: 123-43.
Roth J. The protein A-gold (pAg) technique – a quantitative approach for antigen localization on thin sections. In: Bullock GR, Petrusz P, eds. Techniques in immunocytochemistry. Academic Press, London, New York, 1982; 107-32.
Bendayan M. Protein A-gold electron microscopic immunocytochemistry: methods, applications and limitations. J Electron Microsc Techn. 1984; 1: 243-70.
Reynolds ES. The use of lead citrate at high pH as an electron opaque stain in electron microscopy. J Cell Biol. 1963; 17: 208-12.
Ogoshi M, Inoue K, Takei Y. Identification of a novel adrenomedullin gene family in teleost fish. Biochem Biophys Res Commun. 2003; 311: 1072-74.
Collantes M, Bodegas ME, Sesma MP, Villaro AC. Distribution of proadrenomedullin N-terminal 20 peptide immunoreactivity in the pituitary gland of the frog Rana perezi. Gen Comp Endocrinol. 2003; 1: 50-60.
Martinez A, Unsworth EJ, Cuttitta F. Adrenomedullinlike immunoreactivity in the nervous system of the starfish Marthasterias glacialis. Cell Tissue Res. 1996; 283: 169-72.
Montuenga LM, Martinez A, Miller MJ, Unsworth EJ, Cuttitta F. Expression of adrenomedullin and its receptor during embryogenesis suggests autocrine or paracrine models of action. Endocrinology. 1997; 138: 440-51.
The same distributions of colloidal Au particles were found also in the nerve fibbers penetrating the gland.
Kitani M, Sakata J, Asada Y, Kitamura K, Eto T. Distribution and expression of adrenomedullin in human gastrointestinal tissue. Ann Clin Biochem. 1998; 35: 643-48.
Read_full_article: pdf/22-2012/22-1-2012/SU22-1-2012-Trandaburu.pdf
Correspondence: Ioana Trandaburu, Institute of Biology, Spl. Independenţei 296, 060031, Bucharest, e-mail : itrandaburu@yahoo.com; ioana.trandaburu@ibiol.ro.

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Article Title: The immune-ultrastructural investigations on adrenomedullin (AM) distribution in the pancreas of amphibian Rana esculenta
Authors: Ioana Trandaburu, Wolfgang Kummer, Tiberiu Trandaburu
Affiliation: Center of Cytobiology, Institute of Biology, Splaiul Independenţei 296, 060031 Bucharest, Romania
Institute of Anatomy and Cell Biology, Justus Liebig University, Aulweg 123, 35392 Giessen, Germany
Laboratory of Histology and Embryology, Faculty of Sciences, University of Piteşti, Târgul din Vale 1, 110040 Romania
Abstract: The pancreas of the frog Rana esculenta was investigated in electron microscopy for the immunocytochemical detection of the adrenomedullin (AM). These investigations revealed that AM didn’t label the secretory granules matrix of all endocrine cells, but rather several minority subpopulations of AM, SOM, PP and INS cells. Concretely, the results indicated that 10%-15% from glucagon (GLUC) producing cells, 4%-8% from somatostatin (SOM) and pancreatic polypeptide (PP) cells and only 2%-5% from insulin (INS) cells contained also AM. The sites of antigen-antibody reaction, labelled with colloidal gold particles, have been detected in the cells cytoplasm, randomly distributed among the secretory granules, surrounding them, or overlapped on the hormonal cores. In the exocrine pancreas, reactive nerve fibbers to AM occurred distributed among acini, in the ducts walls, or in those of blood vessels. The above findings and their functional significances are discussed in connection with the results previously reported in mammals and other lower vertebrates.
Keywords: adrenomedullin, AM, frog, pancreas, immunocytochemistry
References: Trandaburu I, Olaru L, Trandaburu T. The occurrence and distribution of adrenomedullin immunoreactivity in the gut of amphibians and reptiles. Proc Inst Biol. 2000; 3: 471-78.
Trandaburu I., Trandaburu T. The occurrence and distribution of adrenomedullin (AM) in the endocrine pancreas of several poikilotherm vertebrates: an immunohistochemical study. Studia Universitatis “Vasile Goldis” Life Sciences. 2010; 20: 27-33.
Kitamura K, Kanagawa K, Kawamoto M, Ichiki Y, Nakamura S, Matsuo H, Eto T. Adrenomedullin: a novel hypotensive peptide isolated from human pheochromocytoma. Biochem Biophys Res Commun. 1993; 192: 533-60.
Wimalawansa SJ. Amylin, calcitonin gene-related peptide, calcitonin, and adrenomedullin: a peptide superfamily. Crit Rev Neurobiol. 1977; 11: 167-239.
Olaru L. Contrbutions regarding the cytoarhitecture and immunohistochemistry of the diffuse neuroendocrine gastro-entero-pancreatic (GEP) system in amphibians. (doctoral thesis), 2005.
Ishiyama Y, Kitamura K, Ichiki Y, Nakamura S, Kida O, Kanagawa K, Eto T. Hemodynamic effects of a novel hypotensive peptide, human adrenomedullin, in rats. Eur J Pharmacol. 1933; 241: 271-73.
Nuki C, Kawasaki H, Kitamura K, Takenaga M, Kanagawa K, Eto T, Wada A. Vasodilator effect of adrenomedullin and calcitonin gene-related peptide receptors in rat mesenteric vascular beds. Biochem Biophys Res Commun. 1993; 196: 245-51.
Allen MA, Ferguson AY. In vitro recordings from area postrema neurons demonstrate responsiveness to adrenomedullin. Am J Physiol. 1996; R 920-R 925.
Kanazawa H, Kurihara N, Hirota K, Kudoh S, Kawaguchi T, Takeda T. Adrenomedullin, a novel discovered hypotensive peptide, is a potent bronchodilator. Biochem Biophys Res Commun. 1994; 205: 251-54.
Miller MJ, Martinez A, Unsworth E, Thiele CJ, Moody TW, Cuttitta F. Adrenomedullin: expression in human tumor cell lines and its potential role as an autocrine growth factor. J Biol Chem. 1996; 271:23345-351.
Walsh TJ, Martinez A, Peter J, Unsworth EJ, Cuttitta F. Antimicrobial activity of adrenomedullin and its gene-related peptide. J Biol Chem. 1998; 273: 3421-428.
Kato F, Kitamura K, Niina H, Yomamoto R, Washimine H, Kanagawa K, Yamamoto Y, Kobayashi H, Eto T, Wada A. Proadrenomedullin N-terminal 20 peptide (PAMP), an endogenous anticholinergic peptide: its exocytotic secretion and inhibition of catecholamine secretion in adrenal medulla. J Neurochem. 1995; 64: 459-61.
Mazzocchi G, Musajo F, Neri G, Gottardo G, Nussdorfer G. Adrenomedullin stimulates steroid secretion by the isolated perfused rat adrenal gland in situ: comparison with calcitonin gene-related peptide effects. Peptides. 1996; 17: 852-57.
Samson WK, Murphy T, Schell DA. A novel vasoactive peptide, adrenomedullin, inhibits pituitary adrenocorticotropin release. Endocrinology. 1995; 136: 2349-352.
Mulder H, Ahrén B, Karlsson S, Sundler F. Adrenomedullin: localization in the gastrointestinal tract and effect on insulin secretion. Regul Pept. 1996; 62: 107-12.
Ishizaka Y, Ishizaka Y, Tanaka M, Kitamura K, Kanagawa K, Minamino N, Matsuo H, Eto T. Adrenomedullin stimulates cyclic AMP formation in rat vascular smooth muscle cells. Biochem Biophys Res Commun. 1994; 200: 642-46.
Washimine H, Asada Y, Kitamura K, Ichiki Y, Hara S, Yamamoto Y, Kanagawa K, Sumiyoshi A, Eto T. Immunohistochemical identification of adrenomedullin in human, rat, and porcine tissues. Histochemistry. 1995; 103: 251-54.
Sakata J, Asada Y, Shimokubo T, Kitani H, Inatsu H, Kitamura K, Kanagawa K, Matsuo H, Sumiyoshi A, Eto T. Adrenomedullin in the gastrointestinal tract. Distribution and gene expression in rat and
augmented gastric adrenomedullin after fasting. J Gastroenterol. 1998; 33: 828-34.
Asada Y, Seiichiro H, Marutsuka K, Kitamura K, Fuji T, Sakata J, Sato J, Kisanuki A, Eto T, Sumiyoshi A. A novel distribution of adrenomedullin immunoreactive cells in human tissues. Cell Tiss Res. 1999; 292: 45-58.
Lopez J, Cuesta N. Adrenomedullin as a pancreatic hormone.Microsc Res Techn. 2002; 57: 61-75.
Marutsuka K, Hatekayama K, Sato Y, Yamashita A, Sumiyoshi A, Asada Y. Immunohistological localization and possible functions of adrenomedullin. Hypertens Res. 2003; 26: 33-40.
Satoh F, Takahashi K, Murakami O, Totsune K, Sone M, Ohneda M, Abe K, Miura J, Hayashi Y, Sasano H, Mouri T. Adrenomedullin in human brain, adrenal glands and tumor tissues of pheochromocytoma, ganglioneuroblastoma and neuroblastoma. J Clin Endocrinol Metabol. 1995; 80: 1750-752.
Pio R, Martinez A, Cuttitta F. Cancer and diabetes: two pathological conditions in which adrenomedullin may be involved. Peptides. 2001; 22: 1719-729.
Gonzales A, Marin O, Sanchez-Camacho C, Pena JJ, Zudaire JJ, Martinez A, Cuttitta F, Munoz M. Localization of adrenomedullin-like immunoreactivity in the hypothalamo-hypophysial system of amphibians. Neurosci Lett. 1998; 242:13-16.
Lopez J, Cuesta N, Cuttitta F, Martinez A. Adrenomedullin in nonmammalian vertebrate pancreas: an immunocytochemical study. Gen Comp Endocrinol. 1999; 3: 309-22.
Zamboni L, De Martino C. Buffered picric acidformaldehyde: a new rapid fixative for electron microscopy. J Cell Biol. 1967; 35: 148A.
Carlemalm E, Garavito RM, Willinger W. Resin development for electron microscopy and an analysis of embedding at low temperature. J Microsc. 1982; 126: 123-43.
Roth J. The protein A-gold (pAg) technique – a quantitative approach for antigen localization on thin sections. In: Bullock GR, Petrusz P, eds. Techniques in immunocytochemistry. Academic Press, London, New York, 1982; 107-32.
Bendayan M. Protein A-gold electron microscopic immunocytochemistry: methods, applications and limitations. J Electron Microsc Techn. 1984; 1: 243-70.
Reynolds ES. The use of lead citrate at high pH as an electron opaque stain in electron microscopy. J Cell Biol. 1963; 17: 208-12.
Ogoshi M, Inoue K, Takei Y. Identification of a novel adrenomedullin gene family in teleost fish. Biochem Biophys Res Commun. 2003; 311: 1072-74.
Collantes M, Bodegas ME, Sesma MP, Villaro AC. Distribution of proadrenomedullin N-terminal 20 peptide immunoreactivity in the pituitary gland of the frog Rana perezi. Gen Comp Endocrinol. 2003; 1: 50-60.
Martinez A, Unsworth EJ, Cuttitta F. Adrenomedullinlike immunoreactivity in the nervous system of the starfish Marthasterias glacialis. Cell Tissue Res. 1996; 283: 169-72.
Montuenga LM, Martinez A, Miller MJ, Unsworth EJ, Cuttitta F. Expression of adrenomedullin and its receptor during embryogenesis suggests autocrine or paracrine models of action. Endocrinology. 1997; 138: 440-51.
The same distributions of colloidal Au particles were found also in the nerve fibbers penetrating the gland.
Kitani M, Sakata J, Asada Y, Kitamura K, Eto T. Distribution and expression of adrenomedullin in human gastrointestinal tissue. Ann Clin Biochem. 1998; 35: 643-48.
*Correspondence: Ioana Trandaburu, Institute of Biology, Spl. Independenţei 296, 060031, Bucharest, e-mail : itrandaburu@yahoo.com; ioana.trandaburu@ibiol.ro.