Summary
In order to examine the intracellular distribution of precursors of type I and type III collagen and fibronectin in the palatal mesenchymal (MEPM) cells of the mouse embryo cultured under ascorbate-deficient conditions, immuno-electron-microscopic studies were carried out by use of affinity purified antibodies for these proteins. MEPM cells were obtained from the palatal shelves of 14-day-old mouse fetuses and cultured for 3–7 days in medium, either with or without 50 ng/dish/day ascorbic acid. Results obtained were as follows: (1) Although the rough endoplasmic reticulum (rER) of MEPM cells cultured for 5 days in ascorbate-supplemented medium was flattened, that in cells cultured in ascorbate-deficient medium had a distended or vesicular appearance. (2) Vesicular or distended rER showed heterogeneous staining for both type I and type III collagen, namely, some parts of rER showed positive staining for both types of collagen, while others showed negative staining. (3) Both type I and type III collagen showed codistribution in the same vesicular rER. (4) Vesicular rER showed negative or very faint labelling for fibronectin. These results may suggest regional differences in the function of rER.
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Blanck TJJ, Peterkofsky B (1975) The stimulation of collagen secretion by ascorbate as a result of increased proline hydroxylation in chick embryo fibroblasts. Arch Biochem Biophys 171:259–267
Chung E, Miller EJ (1974) Collagen polymorphism: characterization of molecules with the chain composition [α1(III)]3 in human tissues. Science 183:1200–1201
Copeland CS, Doms RW, Bolzau EM, Webster RG, Helenius A (1986) Assembly of influenza hemagglutinin trimers and its role in intra-cellular transport. J Cell Biol 103:1179–1191
Danieli JC, Pauli BU, Kuettner KE (1984) Synthesis of cartilage matrix by mammalian chondrocytes in vitro. III. Effects of ascorbate. J Cell Biol 99:1960–1969
Ehrlich HP, Ross R, Bornstein P (1974) Effects of antimicrotubular agents on the secretion of collagen. J Cell Biol 62:390–405
Gething M-J, McCammon K, Sambrook J (1986) Expression of wild-type and mutant forms of influenza hemagglutinin: the role of folding in intracellular transport. Cell 46:939–950
Harwood R, Grant ME, Jackson DS (1976) The route of secretion of procollagen: the influence of α,α′-dipyridyl, colchicine and antimycin A on the secretory process in embryonic-chick tendon and cartilage cells. Biochem J 156:81–90
Hausman E (1967) Cofactor requirements for the enzymatic hydroxylation of lysine in a polypeptide precursor of collagen. Biochem Biophys Acta 133:591–593
Kurisu K, Ohsaki Y, Nagata K, Kukita T, Yoshikawa H, Inai T (1987) Immunoelectron microscopic localization of fibronectin in the smooth muscle layer of mouse small intestine. J Histochem Cytochem 35:411–417
Lodish HF (1988) Transport of secretory and membrane glycoproteins from the rough endoplasmic reticulum to the Golgi. A rate-limiting step in protein maturation and secretion. J Biol Chem 263:2107–2110
Lodish HF, Kong N, Snider M, Strous GJAM (1983) Hepatoma secretory proteins migrate from rough endoplasmic to Golgi at characteristic rates. Nature 304:80–83
Meier S, Solursh M (1978) Ultrastructural analysis of the effect of ascorbic acid on secretion and assembly of extracellular matrix by cultured chick embryo chondrocytes. J Ultrastruct Res 65:48–59
Munro S, Pelham HRB (1987) A C-terminal signal prevents secretion of luminal ER proteins. Cell 48:899–907
Ross R, Benditt EP (1962) Wound healing and collagen formation. II. Fine structure in experimental scurvy. J Cell Biol 12:533–551
Rothman JE (1987) Protein sorting by selective retention in the endoplasmic reticulum and Golgi stack. Cell 50:521–522
Sasaki S, Kurisu K (1983) Effect of triamcinolone acetonide on proliferation and collagen and glycosaminoglycan syntheses in palatal mesenchymal cells from the mouse fetus. J Craniofac Genet Dev Biol 3:351–369
Schafer IA, Silverman L, Sullivan JC, Robertson WVB (1967) Ascorbic acid deficiency in cultured human fibroblast. J Cell Biol 34:83–95
Scheele G, Tartakoff A (1985) Exit of nonglycosylated secretory proteins from the rough endoplasmic reticulum in asynchronous in the exocrine pancreas. J Biol Chem 260:926–931
Slot JW, Geuze HJ (1984) Gold markers for single and double immunolabelling of ultrathin cryosections. In: Polak JM, Varndell IM (eds) Immunolabelling for electron microscopy. Elsevier, Amsterdam, pp 129–142
Slot JW, Geuze HJ (1985) A new method of preparing gold probes for multilabeling cytochemistry. Eur J Cell Biol 38:87–93
Tartakoff AM (1983) Mutations that influence the secretory pathway in animal cells. Biochem J 216:1–9
Thesleff I, Partanen A-M, Kuusela P, Lehtonen E (1987) Dental papilla cells synthesize but do not deposit fibronectin in culture. J Dent Res 66:1107–1115
Timple R, Wick G, Gay S (1977) Antibodies to distinct types of collagens and procollagens and their application in immunohistology. J Immunol Methods 18:165–182
Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354
Tyler MS, Pratt RM (1980) Effect of epidermal growth factor on secondary palatal epithelium in vitro: tissue isolation and recombination studies. J Embryol Exp Morphol 58:93–106
Uitto J, Prockop DJ (1974) Incorporation of proline analogues into collagen polypeptides: effects on the production of extracellular procollagen and on the stability of the triple-helical structure of the molecule. Biochim Biophys Acta 336:234–251
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Kurisu, K., Ohsaki, Y., Nagata, K. et al. Heterogeneous distribution of the precursor of type I and type III collagen and fibronectin in the rough endoplasmic reticulum of palatal mesenchymal cells of the mouse embryo cultured in ascorbate-depleted medium. Cell Tissue Res 267, 429–435 (1992). https://doi.org/10.1007/BF00319365
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DOI: https://doi.org/10.1007/BF00319365