Ultrastructural immunogold study on the various cell types of cultured pancreatic islets of adult rats

• Autorzy: El-Naggar M M
• Języki publikacji: EN

Abstrakty

EN

Whereas several reports describing the ultrastructure of the intact pancreatic islets have been recorded, published experience with the ultrastructural integrity of the cultured pancreatic islets is limited. The present study was, therefore, undertaken to provide an ultrastructure identification of the different cells in the cultured islets of the adult rat pancreas, after marking their secretory granules with gold particles. Pancreatic islets were isolated from adult male Wistar rats by the intraductal perfusion of collagenase technique. The islets were cultured in RPMI-1640 medium for 3 days and processed for preparation of ultrathin sections. The sections were stained with the indirect immunogold technique for insulin, glucagon, somatostatin, and pancreatic polypeptide. Ultrastructural examination of the cultured islets clearly identified the presence of B, A, D and PP-cells, as indicated by the numerous gold particles concentrated predominantly over the secretory granules. The secretory granules of the various cell types of the cultured islets demonstrated several similarities as well as differences from the recorded results of the corresponding secretory granules of the intact islets. The differences probably reflect a deviation in the underlying mechanisms of synthesis, maturation and secretion of the different secretory products of the cells in the cultured islets as they adapt to the in vitro environment.

Słowa kluczowe

EN

electron microscope; pancreas; anatomy; pancreatic islet; rat; adult; ultrastructure; cell type

• Wydawca: -
• Czasopismo: Folia Morphologica
• Rocznik: 2000
• Tom: 59
• Numer: 4
• Opis fizyczny: p.253-262,fig.

Twórcy

autor

El-Naggar M M

• King Abdulaziz University, P.O.Box 9029, Jeddah 21413, Saudi Arabia

Bibliografia

• 1. Buchanan KD, Mawhinney WAA (1973) Glucagon release from isolated pancreas in streptozotocin-treated rats. Diabetes, 22: 797–800.
• 2. Caramia F (1963) Electron microscopic description of a third cell type in the islets of rat pancreas. Am J Anat, 112: 53–64.
• 3. DeMay J, Moremans M, Geuens G, Nuydens R, De Brabander M (1981) High resolution light and electron microscopic localization of tubulin with the IGS (immunogold staining) method. Cell Biol Int Rep, 5: 889–899.
• 4. Dutrillaux MC, Portha B, Rore C, Hollande E (1982) Ultrastructural study of pancreatic B-cell regeneration in newborn rats after destruction by streptozotocin. Virchows Arch Abt B Zellpath, 39: 173–185.
• 5. El-Naggar MM, Elayat AA, Ardawi MSM, Tahir M (1993) Isolated pancreatic islets of the rat: An immunohistochemical and morphometric study. Anat Rec, 237: 489–497.
• 6. Fukuma M (1974) Electron microscopic studies on the granule release from the rat pancreatic B-cells in organ culture. J Electron Microsc, 23: 167–183.
• 7. Fuller A (1969) Elektronenmikroskopische Differenzierung verschiedener Inselzelltypen in Pankreas normaler Albinoratten. Z Zellforsch Mikrosk Anat, 97: 226–248.
• 8. Goldsmith PC, Rose JC, Arimura A, Ganong WF (1975) Ultrastructural localization of somatostatin in pancreatic islets of the rat. Endocrinology, 97: 1061–1064.
• 9. Halban PA, Wolheim CB, Blondel B, Meda P, Neisor EN, Mintz D (1982) The possible importance of contact between pancreatic islet cells for the control of insulin release. Endocrinology, 111: 86–94.
• 10. Hamaguchi K, Nakamura M, Yamaguchi K, Takaki R (1986) Combined effects of glucose and oxygen concentrations upon the ultrastructure of cultured islet cells and insulin accumulation in culture media. Acta Anat, 127: 265–270.
• 11. Howell SL, Fink CJ, Lacy PE (1969a) Isolation and properties of secretory granules from rat islets of Langerhans. J Cell Biol, 41: 154–161.
• 12. Howell SL, Kostianovsky M, Lacy PE (1969b) Beta granule formation in isolated islets of Langerhans. A study by electron microscopic radioautography. J Cell Biol, 42: 692–702.
• 13. Kruse-Jarre D (1982) Secretion in the B cells of islets of Langerhans as demonstrated by zinc staining. Acta diabetol Latino Am, 17: 123–133.
• 14. Lacy PE (1961) Electron microscopy of the B-cell of the pancreas. Amer J Med, 31: 851–859.
• 15. Lacy PE, Kostianovsky M (1967) Method for the isolation of intact islets of Langerhans from the rat pancreas. Diabetes, 16: 35–39.
• 16. Lacy PE, Young DA, Fink CJ (1968) Studies on insulin secretion in vitro from isolated islets of the rat pancreas. Endocrinology, 83: 1155–1161.
• 17. Larsson LI, Sundler F, Hakanson P (1976) Pancreatic polypeptide. A postulate new hormone: identification of its cellular storage site by light and electron microscopic immunocytochemistry. Diabetologia, 12: 211–226.
• 18. Logothotopulos J (1966) Electron microscopy of the pancreatic islets of the rat. Effects of prolonged insulin injections. Diabetes, 15: 823–829.
• 19. Meda P, Michaels RL, Halban PA, Orci L, Sheridan JD (1983). In vivo modulation of gap junctions and dye coupling between B cells of the intact pancreatic islet. Diabetes, 32: 858–868.
• 20. Meyer J, Bencosme SA (1965) The fine structure of normal rabbit pancreatic islet cells. Rev Canad Biol, 24:179–205.
• 21. Moskalewski S (1965) Isolation and culture of the islets of Langerhans of the guinea pig. Gen Comp Endocrinol, 5: 342–353.
• 22. Orci L, lunod A, Pictet R, Renold AE, Rouiller C (1968) Granulolysis in A cells of endocrine pancreas in spontaneous and experimental diabetes in animals. J Cell Biol, 38: 462–466.
• 23. Orci L, Ravazolla MM, Amherdt M, Yanaihara C, Yanaihara N, Alban P, Renold AE, Perrelet A (1984) Insulin, not C peptide (proinsulin), is present in crinophagic bodies of the pancreatic B-cell. J Cell Biol, 98: 222–228.
• 24. Orci L, Unger RH, Renold AE (1973) Structural coupling between pancreatic islet cells. Experientia, 29: 1015–1018.
• 25. Pelletier G (1977) Identification of four cell types in the human endocrine pancreas by immunoelectron microscopy. Diabetes, 26: 749–756.
• 26. Perrier-Barta H (1983) Morphometric study of the secretory process in the endocrine pancreas of the fetal rat. Cell Tissue Res, 229: 651–671.
• 27. Pipeleers D, in’t Veld P, Maes E, Van De Winkel M (1982) Glucose-induced insulin release depends on functional cooperation between islet cells. Proc Natl Acad Sci USA, 79: 7322–7325.
• 28. Sacchi TB, Bani D (1985) New views of the identification of the various cell types in the pancreatic islets of the rat (an ultrastructural and morphometrical study). Acta Anat, 122: 1–17.
• 29. Sacchi TB, Cortesini C, Lombardo DL (1982) The endocrine pancreas of the rat following portacaval shunt. J Submicrosc Cytol, 14: 655–671.
• 30. Slavin BG, Beigelman PM, Bessman SP (1977) Cytophysiological studies on isolated pancreatic islets in vitro. Anat Rec, 188: 445–452.
• 31. Sutton R, Peter M, McShange P, Gray DWR, Morris PJ (1986) An improved method for the isolation of islets of Langerhans from the adult rat pancreas. Transplant Proc, 18: 1819–1820.
• 32. Tahir M, Elayat AA, Jalalah S, El-Naggar MM (1992) Isolated pancreatic islets of the rat: an ultrastructural study. Acta anat, 145: 93–100.
• 33. Vance JE, Buchanan KD, Challoner DR, Williams RH (1968) Effect of glucose concentration on insulin and glucagon release from isolated islets of Langerhans of the rat. Diabetes, 17: 187–193.
• 34. Watson ML (1958) Staining of tissue sections for electron microscopy with heavy metals. J Biophys Biochem Cytol, 4: 475–478.
• 35. Williams MA (1977) Quantitative methods in Biology. In: Glauert AM (ed.) Practical methods in electron microscopy, Vol. 6. North-Holland Publishing Company, Amsterdam, pp: 48–62.
• 36. Williamson JR, Lacy PE, Grisham JW (1961) Ultrastructural changes in the islets of the rat produced by tolbutamide. Diabetes, 10: 460–469.