Abstract
The incorporation of fucose-3H in rat thyroid follicles was studied by radioautography in the light and electron microscopes to determine the site of fucose incorporation into the carbohydrate side chains of thyroglobulin, and to follow the migration of thyroglobulin once it had been labeled with fucose-3H. Radioautographs were examined quantitatively in vivo at several times after injection of fucose-3H into rats, and in vitro following pulse-labeling of thyroid lobes in medium containing fucose-3H. At 3–5 min following fucose-3H administration in vivo, 85% of the silver grains were localized over the Golgi apparatus of thyroid follicular cells. By 20 min, silver grains appeared over apical vesicles, and by 1 hr over the colloid. At 4 hr, nearly all of the silver grains had migrated out of the cells into the colloid. Analysis of the changes in concentration of label with time showed that radioactivity over the Golgi apparatus increased for about 20 min and then decreased, while that over apical vesicles increased to reach a maximum at 35 min. Later, the concentration of label over the apical vesicles decreased, while that over the colloid increased. Similar results were obtained in vitro. It is concluded that fucose, which is located at the end of some of the carbohydrate side chains, is incorporated into thyroglobulin within the Golgi apparatus of thyroid follicular cells, thereby indicating that some of these side chains are completed there. Furthermore, the kinetic analysis demonstrates that apical vesicles are the secretion granules which transport thyroglobulin from the Golgi apparatus to the apex of the cell and release it into the colloid.
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- 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. 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., 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]
- Bouchilloux S., Chabaud O., Michel-Béchet M., Ferrand M., Athouël-Haon A. M. Differential localization in thyroid microsomal subfractions of a mannosyltransferase, two N-acetylglucosaminyltransferases and a galactosyltransferase. Biochem Biophys Res Commun. 1970 Jul 27;40(2):314–320. doi: 10.1016/0006-291x(70)91011-9. [DOI] [PubMed] [Google Scholar]
- Bouchilloux S., Cheftel C. Biosynthesis of thyroglobulin: absence of 14-C-glucosamine incorporation on thyroid polysomes. Biochem Biophys Res Commun. 1966 May 3;23(3):305–310. doi: 10.1016/0006-291x(66)90546-8. [DOI] [PubMed] [Google Scholar]
- Bouchilloux S., Ferrand M., Grégoire J., Chabaud O. Localization in smooth microsomes from sheep thyroid of both a galactosyltransferase and an N-acetylhexosaminyltransferase. Biochem Biophys Res Commun. 1969 Oct 22;37(3):538–544. doi: 10.1016/0006-291x(69)90949-8. [DOI] [PubMed] [Google Scholar]
- CARO L. G. High-resolution autoradiogaphy. II. The problem of resolution. J Cell Biol. 1962 Nov;15:189–199. doi: 10.1083/jcb.15.2.189. [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]
- Coimbra A., Leblond C. P. Sites of glycogen synthesis in rat liver cells as shown by electron microscope radioautography after administration of glucose-H3. J Cell Biol. 1966 Jul;30(1):151–175. doi: 10.1083/jcb.30.1.151. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Droz B. Elaboration de glycoproteines dans l'appareil de Golgi des cellules hépatiques chez le rat; étude radioautographique en microscopie électronique après injection de glactose-3H. C R Acad Sci Hebd Seances Acad Sci D. 1966 Apr 18;262(16):1766–1768. [PubMed] [Google Scholar]
- Gottschalk A. Biosynthesis of glycoproteins and its relationship to heterogeneity. Nature. 1969 May 3;222(5192):452–454. doi: 10.1038/222452a0. [DOI] [PubMed] [Google Scholar]
- Herscovics A. Biosynthesis of thyroglobulin. Incorporation of [1-14C] galactose, [1-14C] manose and [4,5-3H2] leucine into soluble proteins by rat thyroids in vitro. Biochem J. 1969 May;112(5):709–719. doi: 10.1042/bj1120709. [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]
- JENSEN F. C., GWATKIN R. B., BIGGERS J. D. A SIMPLE ORGAN CULTURE METHOD WHICH ALLOWS SIMULTANEOUS ISOLATION OF SPECIFIC TYPES OF CELLS. Exp Cell Res. 1964 May;34:440–447. doi: 10.1016/0014-4827(64)90231-9. [DOI] [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]
- NADLER N. J., SARKAR S. K., LEBLOND C. P. Origin of intracellular colloid droplets in the rat thyroid. Endocrinology. 1962 Jul;71:120–129. doi: 10.1210/endo-71-1-120. [DOI] [PubMed] [Google Scholar]
- NADLER N. J., YOUNG B. A., LEBLOND C. P., MITMAKER B. ELABORATION OF THYROGLOBULIN IN THE THYROID FOLLICLE. Endocrinology. 1964 Mar;74:333–354. doi: 10.1210/endo-74-3-333. [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]
- Neutra M., Leblond C. P. Synthesis of the carbohydrate of mucus in the golgi complex as shown by electron microscope radioautography of goblet cells from rats injected with glucose-H3. J Cell Biol. 1966 Jul;30(1):119–136. doi: 10.1083/jcb.30.1.119. [DOI] [PMC free article] [PubMed] [Google Scholar]
- PETERSON M., LEBLOND C. P. SYNTHESIS OF COMPLEX CARBOHYDRATES IN THE GOLGI REGION, AS SHOWN BY RADIOAUTOGRAPHY AFTER INJECTION OF LABELED GLUCOSE. J Cell Biol. 1964 Apr;21:143–148. doi: 10.1083/jcb.21.1.143. [DOI] [PMC free article] [PubMed] [Google Scholar]
- SPIRO R. G., SPIRO M. J. THE CARBOHYDRATE COMPOSITION OF THE THYROGLOBULINS FROM SEVERAL SPECIES. J Biol Chem. 1965 Mar;240:997–1001. [PubMed] [Google Scholar]
- SPIRO R. G. THE CARBOHYDRATE UNITS OF THYROGLOBULIN. J Biol Chem. 1965 Apr;240:1603–1610. [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: their biochemistry, biology and role in human disease (first of two parts). N Engl J Med. 1969 Oct 30;281(18):991–contd. doi: 10.1056/NEJM196910302811806. [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]
- WISSIG S. L. The anatomy of secretion in the follicular cells of the thyroid gland. I. The fine structure of the gland in the normal rat. J Biophys Biochem Cytol. 1960 Jun;7:419–432. doi: 10.1083/jcb.7.3.419. [DOI] [PMC free article] [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]