Abstract
A single intravenous injection of L-[3H]fucose, a specific glycoprotein precursor, was given to young 35–45 g rats which were sacrificed at times varying between 2 min and 30 h later. Radioautography of over 50 cell types, including renewing and nonrenewing cells, was carried out for light and electron microscope study. At early time intervals (2–10 min after injection), light microscope radioautography showed a reaction over nearly all cells investigated in the form of a discrete clump of silver grains over the Golgi region. This reaction varied in intensity and duration from cell type to cell type. Electron microscope radioautographs of duodenal villus columnar cells and kidney proximal and distal tubule cells at early time intervals revealed that the silver grains were restricted to Golgi saccules. These observations are interpreted to mean that glycoproteins undergoing synthesis incorporate fucose in the saccules of the Golgi apparatus. Since fucose occurs as a terminal residue in the carbohydrate side chains of glycoproteins, the Golgi saccules would be the site of completion of synthesis of these side chains. At later time intervals, light and electron microscope radioautography demonstrated a decrease in the reaction intensity of the Golgi region, while reactions appeared over other parts of the cells: lysosomes, secretory material, and plasma membrane. The intensity of the reactions observed over the plasma membrane varied considerably in various cell types; furthermore the reactions were restricted to the apical surface in some types, but extended to the whole surface in others. Since the plasma membrane is covered by a "cell coat" composed of the carbohydrate-rich portions of membrane glycoproteins, it is concluded that newly formed glycoproteins, after acquiring fucose in the Golgi apparatus, migrate to the cell surface to contribute to the cell coat. This contribution implies turnover of cell coat glycoproteins, at least in nonrenewing cell types, such as those of kidney tubules. In the young cells of renewing populations, e.g. those of gastro-intestinal epithelia, the new glycoproteins seem to contribute to the growth as well as the turnover of the cell coat. The differences in reactivity among different cell types and cell surfaces imply considerable differences in the turnover rates of the cell coats.
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- Alpers D. H. The relation of size to the relative rates of degradation of intestinal brush border proteins. J Clin Invest. 1972 Oct;51(10):2621–2630. doi: 10.1172/JCI107080. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bekesi J. G., Winzler R. J. The metabolism of plasma glycoproteins. Studies on the incorporation of L-fucose-1-14-C into tissue and serum in the normal rat. J Biol Chem. 1967 Sep 10;242(17):3873–3879. [PubMed] [Google Scholar]
- Bell D. J., Taluker M. Q. Fucose in urine of fasting human subjects. Biochem J. 1971 Mar;122(1):24P–24P. doi: 10.1042/bj1220024pa. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Benedetti E. L., Emmelot P. Studies on plasma membranes. IV. The ultrastructural localization and content of sialic acid in plasma membranes isolated from rat liver and hepatoma. J Cell Sci. 1967 Dec;2(4):499–512. doi: 10.1242/jcs.2.4.499. [DOI] [PubMed] [Google Scholar]
- Bennett G., Leblond C. P. Formation of cell coat material for the whole surface of columnar cells in the rat small intestine, as visualized by radioautography with L-fucose-3H. J Cell Biol. 1970 Aug;46(2):409–416. doi: 10.1083/jcb.46.2.409. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bennett G., Leblond C. P. Passage of fucose- 3 H label from the Golgi apparatus into dense and multivesicular bodies in the duodenal columnar cells and hepatocytes of the rat. J Cell Biol. 1971 Dec;51(3):875–881. doi: 10.1083/jcb.51.3.875. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bennett G. Migration of glycoprotein from golgi apparatus to cell coat in the columnar cells of the duodenal epithelium. J Cell Biol. 1970 Jun;45(3):668–673. doi: 10.1083/jcb.45.3.668. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bocci V., Winzler R. J. Metabolism of L-fucose-1-14C and of fucose glycoproteins in the rat. Am J Physiol. 1969 Jun;216(6):1337–1342. doi: 10.1152/ajplegacy.1969.216.6.1337. [DOI] [PubMed] [Google Scholar]
- Bosmann H. B. Cell surface glycosyl transferases and acceptors in normal and RNA- and DNA-virus transformed fibroblasts. Biochem Biophys Res Commun. 1972 Aug 7;48(3):523–529. doi: 10.1016/0006-291x(72)90379-8. [DOI] [PubMed] [Google Scholar]
- Bosmann H. B., Hagopian A., Eylar E. H. Cellular membranes: the biosynthesis of glycoprotein and glycolipid in hela cell membranes. Arch Biochem Biophys. 1969 Mar;130(1):573–583. doi: 10.1016/0003-9861(69)90073-3. [DOI] [PubMed] [Google Scholar]
- Bosmann H. B., Hagopian A., Eylar E. H. Glycoprotein biosynthesis: the characterization of two glycoprotein:frucosyl transferases in HeLa cells. Arch Biochem Biophys. 1968 Nov;128(2):470–481. doi: 10.1016/0003-9861(68)90053-2. [DOI] [PubMed] [Google Scholar]
- CHOI J. K. The fine structure of the urinary bladder of the toad, Bufo marinus. J Cell Biol. 1963 Jan;16:53–72. doi: 10.1083/jcb.16.1.53. [DOI] [PMC free article] [PubMed] [Google Scholar]
- COFFEY J. W., MILLER O. N., SELLINGER O. Z. THE METABOLISM OF L-FUCOSE IN THE RAT. J Biol Chem. 1964 Dec;239:4011–4017. [PubMed] [Google Scholar]
- Forstner G. G. Release of intestinal surface-membrane glycoproteins associated with enzyme activity by brief digestion with papain. Biochem J. 1971 Mar;121(5):781–789. doi: 10.1042/bj1210781. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Forstner G. G. Surface sugar in the intestine. Am J Med Sci. 1969 Sep;258(3):172–180. doi: 10.1097/00000441-196909000-00004. [DOI] [PubMed] [Google Scholar]
- GLEGG R. E., EIDINGER D., LEBLOND C. P. Presence of carbohydrates distinct from acid mucopolysaccharides in connective tissue. Science. 1954 Nov 19;120(3125):839–840. doi: 10.1126/science.120.3125.839. [DOI] [PubMed] [Google Scholar]
- Glossmann H., Neville D. M., Jr Glycoproteins of cell surfaces. A comparative study of three different cell surfaces of the rat. J Biol Chem. 1971 Oct 25;246(20):6339–6346. [PubMed] [Google Scholar]
- Glossmann H., Neville D. M. gamma-Glutamyltransferase in kidney brush border membranes. FEBS Lett. 1972 Jan 1;19(4):340–344. doi: 10.1016/0014-5793(72)80075-9. [DOI] [PubMed] [Google Scholar]
- Goldstone A., Koenig H. Lysosomal hydrolases as glycoproteins. Life Sci II. 1970 Dec 8;9(23):1341–1350. doi: 10.1016/0024-3205(70)90115-3. [DOI] [PubMed] [Google Scholar]
- Haddad A., Smith M. D., Herscovics A., Nadler N. J., Leblond C. P. Radioautographic study of in vivo and in vitro incorporation of fucose-3H into thyroglobulin by rat thyroid follicular cells. J Cell Biol. 1971 Jun;49(3):856–877. doi: 10.1083/jcb.49.3.856. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Herscovics A. Biosynthesis of thyroglobulin: incorporation of [3H] fucose into proteins by rat thyroids in vitro. Biochem J. 1970 Apr;117(2):411–413. doi: 10.1042/bj1170411. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hughes R. C., Sanford B., Jeanloz R. W. Regeneration of the surface glycoproteins of a transplantable mouse tumor cell after treatment with neuraminidase. Proc Natl Acad Sci U S A. 1972 Apr;69(4):942–945. doi: 10.1073/pnas.69.4.942. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ito S. Structure and function of the glycocalyx. Fed Proc. 1969 Jan-Feb;28(1):12–25. [PubMed] [Google Scholar]
- Kaufman R. L., Ginsburg V. The metabolism of L-fucose by HeLa cells. Exp Cell Res. 1968 Apr;50(1):127–132. doi: 10.1016/0014-4827(68)90400-x. [DOI] [PubMed] [Google Scholar]
- Kraemer P. M. Regeneration of sialic acid on the surface of Chinese hamster cells in culture. II. Incorporation of radioactivity from glucosamine-1-14C. J Cell Physiol. 1967 Apr;69(2):199–207. doi: 10.1002/jcp.1040690210. [DOI] [PubMed] [Google Scholar]
- Kraemer P. M., Tobey R. A. Cell-cycle dependent desquamation of heparan sulfate from the cell surface. J Cell Biol. 1972 Dec;55(3):713–717. doi: 10.1083/jcb.55.3.713. [DOI] [PMC free article] [PubMed] [Google Scholar]
- LEBLOND C. P. CLASSIFICATION OF CELL POPULATIONS ON THE BASIS OF THEIR PROLIFERATIVE BEHAVIOR. Natl Cancer Inst Monogr. 1964 May;14:119–150. [PubMed] [Google Scholar]
- LEBLOND C. P. Distribution of periodic acid-reactive carbohydrates in the adult rat. Am J Anat. 1950 Jan;86(1):1–49. doi: 10.1002/aja.1000860102. [DOI] [PubMed] [Google Scholar]
- LEBLOND C. P., GLEGG R. E., EIDINGER D. Presence of carbohydrates with free 1,2-glycol groups in sites stained by the periodic acid-Schiff technique. J Histochem Cytochem. 1957 Sep;5(5):445–458. doi: 10.1177/5.5.445. [DOI] [PubMed] [Google Scholar]
- Marchesi V. T., Tillack T. W., Jackson R. L., Segrest J. P., Scott R. E. Chemical characterization and surface orientation of the major glycoprotein of the human erythrocyte membrane. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1445–1449. doi: 10.1073/pnas.69.6.1445. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maury P. Neuraminyl-oligosaccharides of rat urine: isolation and some structural characteristics. Biochim Biophys Acta. 1971 Oct;252(1):48–57. doi: 10.1016/0304-4165(71)90091-2. [DOI] [PubMed] [Google Scholar]
- Molnar J., Teegraden D. W., Winzler R. J. The biosynthesis of glycoproteins. VI. Production of extracellular radioactive macromolecules by Ehrlich ascites carcinoma cells during incubation with glucosamine-14C. Cancer Res. 1965 Dec;25(11):1860–1866. [PubMed] [Google Scholar]
- Morre J., Merlin L. M., Keenan T. W. Localization of glycosyl transferase activities in a Golgi apparatus-rich fraction isolated from rat liver. Biochem Biophys Res Commun. 1969 Nov 20;37(5):813–819. doi: 10.1016/0006-291x(69)90964-4. [DOI] [PubMed] [Google Scholar]
- Neutra M., Leblond C. P. Radioautographic comparison of the uptake of galactose-H and glucose-H3 in the golgi region of various cells secreting glycoproteins or mucopolysaccharides. J Cell Biol. 1966 Jul;30(1):137–150. doi: 10.1083/jcb.30.1.137. [DOI] [PMC free article] [PubMed] [Google Scholar]
- PUCHTLER H., LEBLOND C. P. Histochemical analysis of cell membranes and associated structures as seen in the intestinal epithelium. Am J Anat. 1958 Jan;102(1):1–31. doi: 10.1002/aja.1001020102. [DOI] [PubMed] [Google Scholar]
- Pelletier G., Puviani R. Detection of glycoproteins and autoradiographic localization of ( 3 H)fucose in the thyroidectomy cells of rat anterior pituitary gland. J Cell Biol. 1973 Feb;56(2):600–605. doi: 10.1083/jcb.56.2.600. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rambourg A., Hernandez W., Leblond C. P. Detection of complex carbohydrates in the Golgi apparatus of rat cells. J Cell Biol. 1969 Feb;40(2):395–414. doi: 10.1083/jcb.40.2.395. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rambourg A., Leblond C. P. Electron microscope observations on the carbohydrate-rich cell coat present at the surface of cells in the rat. J Cell Biol. 1967 Jan;32(1):27–53. doi: 10.1083/jcb.32.1.27. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rambourg A. Morphological and histochemical aspects of glycoproteins at the surface of animal cells. Int Rev Cytol. 1971;31:57–114. doi: 10.1016/s0074-7696(08)60057-1. [DOI] [PubMed] [Google Scholar]
- Rambourg A., Neutra M., Leblond C. P. Presence of a "cell coat" rich in carbohydrate at the surface of cells in the rat. Anat Rec. 1966 Jan;154(1):41–71. doi: 10.1002/ar.1091540105. [DOI] [PubMed] [Google Scholar]
- Roseman S. The synthesis of complex carbohydrates by multiglycosyltransferase systems and their potential function in intercellular adhesion. Chem Phys Lipids. 1970 Oct;5(1):270–297. doi: 10.1016/0009-3084(70)90024-1. [DOI] [PubMed] [Google Scholar]
- Roth S., McGuire E. J., Roseman S. Evidence for cell-surface glycosyltransferases. Their potential role in cellular recognition. J Cell Biol. 1971 Nov;51(21):536–547. doi: 10.1083/jcb.51.2.536. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schachter H., Jabbal I., Hudgin R. L., Pinteric L., McGuire E. J., Roseman S. Intracellular localization of liver sugar nucleotide glycoprotein glycosyltransferases in a Golgi-rich fraction. J Biol Chem. 1970 Mar 10;245(5):1090–1100. [PubMed] [Google Scholar]
- Singer S. J., Nicolson G. L. The fluid mosaic model of the structure of cell membranes. Science. 1972 Feb 18;175(4023):720–731. doi: 10.1126/science.175.4023.720. [DOI] [PubMed] [Google Scholar]
- Smeds S. Protein composition of the colloid collected from single rat thyroid follicles. Biochem Biophys Res Commun. 1970 Mar 27;38(6):1168–1173. doi: 10.1016/0006-291x(70)90362-1. [DOI] [PubMed] [Google Scholar]
- Spiro R. G. Glycoproteins. Annu Rev Biochem. 1970;39:599–638. doi: 10.1146/annurev.bi.39.070170.003123. [DOI] [PubMed] [Google Scholar]
- Spiro R. G., Spiro M. J. Glycoprotein biosynthesis: studies on thyroglobulin. Characterization of a particulate precursor and radioisotope incorporation by thyroid slices and particle systems. J Biol Chem. 1966 Mar 25;241(6):1271–1282. [PubMed] [Google Scholar]
- Warnes T. W. Alkaline phosphatase. Gut. 1972 Nov;13(11):926–937. doi: 10.1136/gut.13.11.926. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Warren L., Glick M. C. Membranes of animal cells. II. The metabolism and turnover of the surface membrane. J Cell Biol. 1968 Jun;37(3):729–746. doi: 10.1083/jcb.37.3.729. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Warshawsky H., Moore G. A technique for the fixation and decalcification of rat incisors for electron microscopy. J Histochem Cytochem. 1967 Sep;15(9):542–549. doi: 10.1177/15.9.542. [DOI] [PubMed] [Google Scholar]
- Weinstock A., Weinstock M., Leblond C. P. Autoradiographic detection of 3 H-fucose incorporation into glycoprotein by odontoblasts and its deposition at the site of the calcification front in dentin. Calcif Tissue Res. 1972;8(3):181–189. doi: 10.1007/BF02010136. [DOI] [PubMed] [Google Scholar]
- Whur P., Herscovics A., Leblond C. P. Radioautographic visualization of the incorporation of galactose-3H and mannose-3H by rat thyroids in vitro in relation to the stages of thyroglobulin synthesis. J Cell Biol. 1969 Nov;43(2):289–311. doi: 10.1083/jcb.43.2.289. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Winzler R. J. Carbohydrates in cell surfaces. Int Rev Cytol. 1970;29:77–125. doi: 10.1016/s0074-7696(08)60033-9. [DOI] [PubMed] [Google Scholar]