Abstract
The pathway and kinetics of the secretory protein transport in rat lacrimal exorbital gland have been established by an in vitro time- course radioautographic study of pulse-labeled protein secretion. The colchicine-sensitive steps have been localized by using the drug at various times with respect to the pulse labeling of proteins. Colchicine (10 microM) does not block any step of the secretory protein transport, but when introduced before the pulse it decreases the transfer of labeled proteins from the rough endoplasmic reticulum to the Golgi area, suppressing their temporary accumulation in the Golgi area before any alteration of this organelle is detectable. Moreover, colchicine inhibits protein release only from the secretory granules formed in its presence because the peroxidase discharge is diminished 1 h after colchicine addition, and the secretion of newly synthesized proteins is strongly inhibited only when colchicine is introduced before secretory granule formation. Morphometric studies show that there is a great increase of secondary lysosomes, related to crinophagy, as early as 40-50 min after colchicine is added. However, changes in lysosomal enzymatic activities remained biochemically undetectable. We conclude that: (a) the labile microtubular system does not seem indispensable for protein transport in the rough endoplasmic reticulum-Golgi area but may facilitate this step, perhaps by maintaining the spatial organization of this area; and (b) in the lacrimal gland, colchicine inhibits protein release not by acting on the steps of secretion following the secretory granule formation, but by acting chiefly on the steps preceding secretory granule formation, perhaps by making the secretory granules formed in its presence incapable of discharging their content.
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- Antakly T., Pelletier G., Zeytinoglu F., Labrie F. Effects of colchicine on the morphology and prolactin secretion of rat anterior pituitary cells in monolayer culture. Am J Anat. 1979 Nov;156(3):353–371. doi: 10.1002/aja.1001560306. [DOI] [PubMed] [Google Scholar]
- Ashley C. A., Peters T., Jr Electron microscopic radioautographic detection of sites of protein synthesis and migration in liver. J Cell Biol. 1969 Nov;43(2):237–249. doi: 10.1083/jcb.43.2.237. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Banerjee D., Manning C. P., Redman C. M. The in vivo effect of colchicine on the addition of galactose and sialic acid to rat hepatic serum glycoproteins. J Biol Chem. 1976 Jul 10;251(13):3887–3892. [PubMed] [Google Scholar]
- Blok J., Ginsel L. A., Mulder-Stapel A. A., Onderwater J. J., Daems W. T. The effect of colchicine on the intracellular transport of 3H-fucose-labelled glycoproteins in the absorptive cells of cultured human small-intestinal tissue. An autoradiographical and biochemical study. Cell Tissue Res. 1981;215(1):1–12. doi: 10.1007/BF00236244. [DOI] [PubMed] [Google Scholar]
- Castle J. D., Jamieson J. D., Palade G. E. Radioautographic analysis of the secretory process in the parotid acinar cell of the rabbit. J Cell Biol. 1972 May;53(2):290–311. doi: 10.1083/jcb.53.2.290. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chambaut-Guérin A. M., Muller P., Rossignol B. Microtubules and protein secretion in rat lacrimal glands. Inhibitory effect of the tubulin . colchicine complex isolated from lacrimal glands upon brain tubulin polymerization. Identification of the complex by gel electrophoresis. J Biol Chem. 1979 Nov 10;254(21):10734–10739. [PubMed] [Google Scholar]
- Chambaut-Guérin A. M., Muller P., Rossignol B. Microtubules and protein secretion in rat lacrimal glands. Relationship between colchicine binding and its inhibitory effect on the intracellular transport of proteins. J Biol Chem. 1978 Jun 10;253(11):3870–3876. [PubMed] [Google Scholar]
- Chu L. L., MacGregor R. R., Cohn D. V. Energy-dependent intracellular translocation of proparathormone. J Cell Biol. 1977 Jan;72(1):1–10. doi: 10.1083/jcb.72.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Diegelmann R. F., Peterkofsky B. Inhibition of collagen secretion from bone and cultured fibroblasts by microtubular disruptive drugs. Proc Natl Acad Sci U S A. 1972 Apr;69(4):892–896. doi: 10.1073/pnas.69.4.892. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ehrlich H. P., Ross R., Bornstein P. Effects of antimicrotubular agents on the secretion of collagen. A biochemical and morphological study. J Cell Biol. 1974 Aug;62(2):390–405. doi: 10.1083/jcb.62.2.390. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Essner E. Localization of endogenous peroxidase in rat exorbital lacrimal gland. J Histochem Cytochem. 1971 Apr;19(4):216–225. doi: 10.1177/19.4.216. [DOI] [PubMed] [Google Scholar]
- Feldmann G., Maurice M., Sapin C., Benhamou J. P. Inhibition by colchicine of fibrinogen translocation in hepatocytes. J Cell Biol. 1975 Oct;67(1):237–243. doi: 10.1083/jcb.67.1.237. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Flickinger C. J. Synthesis, intracellular transport, and release of secretory protein in the seminal vesicle of the rat, as studied by electron microscope radioautography. Anat Rec. 1974 Nov;180(3):407–425. doi: 10.1002/ar.1091800302. [DOI] [PubMed] [Google Scholar]
- Glaumann H., Ericsson J. L. Evidence for the participation of the Golgi apparatus in the intracellular transport of nascent albumin in the liver cell. J Cell Biol. 1970 Dec;47(3):555–567. doi: 10.1083/jcb.47.3.555. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Habener J. F., Amherdt M., Ravazzola M., Orci L. Parathyroid hormone biosynthesis. Correlation of conversion of biosynthetic precursors with intracellular protein migration as determined by electron microscope autoradiography. J Cell Biol. 1979 Mar;80(3):715–731. doi: 10.1083/jcb.80.3.715. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Habener J. F., Kemper B., Potts J. T., Jr Calcium-dependent intracellular degradation of parathyroid hormone: a possible mechanism for the regulation of hormone stores. Endocrinology. 1975 Aug;97(2):431–441. doi: 10.1210/endo-97-2-431. [DOI] [PubMed] [Google Scholar]
- Herzog V., Sies H., Miller F. Exocytosis in secretory cells of rat lacrimal gland. Peroxidase release from lobules and isolated cells upon cholinergic stimulation. J Cell Biol. 1976 Sep;70(3):692–706. doi: 10.1083/jcb.70.3.692. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hoffstein S., Goldstein I. M., Weissmann G. Role of microtubule assembly in lysosomal enzyme secretion from human polymorphonuclear leukocytes. A reevaluation. J Cell Biol. 1977 Apr;73(1):242–256. doi: 10.1083/jcb.73.1.242. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Howell S. L., Tyhurst M. Role of microtubules in the intracellular transport of growth hormone. Cell Tissue Res. 1978 Jun 26;190(1):163–171. doi: 10.1007/BF00210045. [DOI] [PubMed] [Google Scholar]
- Jamieson J. D., Palade G. E. Intracellular transport of secretory proteins in the pancreatic exocrine cell. II. Transport to condensing vacuoles and zymogen granules. J Cell Biol. 1967 Aug;34(2):597–615. doi: 10.1083/jcb.34.2.597. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kanagasuntheram P., Lim S. C. parallel secretion of secretory proteins and calcium by the rat parotid gland. J Physiol. 1981 Mar;312:445–454. doi: 10.1113/jphysiol.1981.sp013637. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kemper B., Habener J. F., Rich A., Potts J. T., Jr Microtubules and the intracellular conversion of proparathyroid hormone to parathyroid hormone. Endocrinology. 1975 Apr;96(4):903–912. doi: 10.1210/endo-96-4-903. [DOI] [PubMed] [Google Scholar]
- Knudson C. M., Stemberger B. H., Patton S. Effects of colchicine on ultrastructure of the lactating mammary cell: membrane involvement and stress on the Golgi apparatus. Cell Tissue Res. 1978 Dec 14;195(1):169–181. doi: 10.1007/BF00233684. [DOI] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Le Marchand Y., Singh A., Assimacopoulos-Jeannet F., Orci L., Rouiller C., Jeanrenaud B. A role for the microtubular system in the release of very low density lipoproteins by perfused mouse livers. J Biol Chem. 1973 Oct 10;248(19):6862–6870. [PubMed] [Google Scholar]
- Lohmander S., Moskalewski S., Madsen K., Thyberg J., Friberg U. Influence of colchicine on the synthesis and secretion of proteoglycans and collagen by fetal guinea pig chondrocytes. Exp Cell Res. 1976 May;99(2):333–345. doi: 10.1016/0014-4827(76)90591-7. [DOI] [PubMed] [Google Scholar]
- Malaisse-Lagae F., Amherdt M., Ravazzola M., Sener A., Hutton J. C., Orci L., Malaisse W. J. Role of microtubules in the synthesis, conversion, and release of (pro)insulin. A biochemical and radioautographic study in rat islets. J Clin Invest. 1979 Jun;63(6):1284–1296. doi: 10.1172/JCI109423. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Malaisse W. J., Malaisse-Lagae F., Van Obberghen E., Somers G., Devis G., Ravazzola M., Orci L. Role of microtubules in the phasic pattern of insulin release. Ann N Y Acad Sci. 1975 Jun 30;253:630–652. doi: 10.1111/j.1749-6632.1975.tb19234.x. [DOI] [PubMed] [Google Scholar]
- Malaisse W. J., Orci L. The role of the cytoskeleton in pancreatic B-cell function. Methods Achiev Exp Pathol. 1979;9:112–136. [PubMed] [Google Scholar]
- Moskalewski S., Thyberg J., Friberg U. In vitro influence of colchicine on the Golgi complex in A- and B-cells of guinea pig pancreatic islets. J Ultrastruct Res. 1976 Feb;54(2):304–317. doi: 10.1016/s0022-5320(76)80159-1. [DOI] [PubMed] [Google Scholar]
- Moskalewski S., Thyberg J., Lohmander S., Friberg U. Influence of colchicine and vinblastine on the golgi complex and matrix deposition in chondrocyte aggregates. An ultrastructural study. Exp Cell Res. 1975 Oct 15;95(2):440–454. doi: 10.1016/0014-4827(75)90569-8. [DOI] [PubMed] [Google Scholar]
- Nadler N. J. Quantitation and resolution in electron microscope radioautography. J Histochem Cytochem. 1979 Nov;27(11):1531–1533. doi: 10.1177/27.11.512340. [DOI] [PubMed] [Google Scholar]
- Nickerson S. C., Smith J. J., Keenan T. W. Role of microtubules in milk secretion--action of colchicine on microtubules and exocytosis of secretory vesicles in rat mammary epithelial cells. Cell Tissue Res. 1980;207(3):361–376. doi: 10.1007/BF00224613. [DOI] [PubMed] [Google Scholar]
- Ollivier-Bousquet M., Denamur R. Inhibition par la colchicine de la sécrétion des protéines du lait. C R Acad Sci Hebd Seances Acad Sci D. 1973 Apr 2;276(14):2183–2186. [PubMed] [Google Scholar]
- Orci L., Le Marchand Y., Singh A., Assimacopoulos-Jeannet F., Rouiller C., Jeanrenaud B. Letter: Role of microtubules in lipoprotein secretion by the liver. Nature. 1973 Jul 6;244(5410):30–32. doi: 10.1038/244030a0. [DOI] [PubMed] [Google Scholar]
- Orci L., Like A. A., Amherdt M., Blondel B., Kanazawa Y., Marliss E. B., Lambert A. E., Wollheim C. B., Renold A. E. Monolayer cell culture of neonatal rat pancreas: an ultrastructural and biochemical study of functioning endocrine cells. J Ultrastruct Res. 1973 May;43(3):270–297. doi: 10.1016/s0022-5320(73)80039-5. [DOI] [PubMed] [Google Scholar]
- Palade G. Intracellular aspects of the process of protein synthesis. Science. 1975 Aug 1;189(4200):347–358. doi: 10.1126/science.1096303. [DOI] [PubMed] [Google Scholar]
- Patzelt C., Brown D., Jeanrenaud B. Inhibitory effect of colchicine on amylase secretion by rat parotid glands. Possible localization in the Golgi area. J Cell Biol. 1977 Jun;73(3):578–593. doi: 10.1083/jcb.73.3.578. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pelletier G., Bornstein M. B. Effect of colchicine on rat anterior pituitary gland in tissue culture. Exp Cell Res. 1972 Jan;70(1):221–223. doi: 10.1016/0014-4827(72)90200-5. [DOI] [PubMed] [Google Scholar]
- Piasek A., Thyberg J. Effects of colchicine on endocytosis and cellular inactivation of horseradish peroxidase in cultured chondrocytes. J Cell Biol. 1979 May;81(2):426–437. doi: 10.1083/jcb.81.2.426. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Piasek A., Thyberg J. Effects of colchicine on endocytosis of horseradish peroxidase by rat peritoneal macrophages. J Cell Sci. 1980 Oct;45:59–71. doi: 10.1242/jcs.45.1.59. [DOI] [PubMed] [Google Scholar]
- Putney J. W., Jr, VandeWalle C. M., Leslie B. A. Stimulus-secretion coupling in the rat lacrimal gland. Am J Physiol. 1978 Nov;235(5):C188–C198. doi: 10.1152/ajpcell.1978.235.5.C188. [DOI] [PubMed] [Google Scholar]
- ROBBINS E., GONATAS N. K. HISTOCHEMICAL AND ULTRASTRUCTURAL STUDIES ON HELA CELL CULTURES EXPOSED TO SPINDLE INHIBITORS WITH SPECIAL REFERENCE TO THE INTERPHASE CELL. J Histochem Cytochem. 1964 Sep;12:704–711. doi: 10.1177/12.9.704. [DOI] [PubMed] [Google Scholar]
- Reaven E. P., Reaven G. M. Dissociation between rate of hepatic lipoprotein secretion and hepatocyte microtubule content. J Cell Biol. 1978 Jun;77(3):735–742. doi: 10.1083/jcb.77.3.735. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Redman C. M., Banerjee D., Howell K., Palade G. E. Colchicine inhibition of plasma protein release from rat hepatocytes. J Cell Biol. 1975 Jul;66(1):42–59. doi: 10.1083/jcb.66.1.42. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Redman C. M., Banerjee D., Manning C., Huang C. Y., Green K. In vivo effect of colchicine on hepatic protein synthesis and on the conversion of proalbumin to serum albumin. J Cell Biol. 1978 May;77(2):400–416. doi: 10.1083/jcb.77.2.400. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rohr H. P., Schmalbeck J., Feldhege A. Elektronenmikroskopisch-autoradiographische Untersuchungen über Eiweiss-Synthese in der Brunnerschen Drüse der Maus. Z Zellforsch Mikrosk Anat. 1967;80(2):183–204. [PubMed] [Google Scholar]
- Rohr H., Seitter U., Schmalbeck J. Voraussetzungen und derzeitige Grenzen der quanitiativen elektronenmikroskopischen Autoradiographie bei Kinetikstudien an Drüsenzellen. Elektronenmikroskopish-autoradiographische Untersuchungen mit 3H-Leucin an der Milchdrüse. Z Zellforsch Mikrosk Anat. 1968;85(3):376–397. [PubMed] [Google Scholar]
- SELLINGER O. Z., BEAUFAY H., JACQUES P., DOYEN A., DE DUVE C. Tissue fractionation studies. 15. Intracellular distribution and properties of beta-N-acetylglucosaminidase and beta-galactosidase in rat liver. Biochem J. 1960 Mar;74:450–456. doi: 10.1042/bj0740450. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Scheele G. A. Biosynthesis, segregation, and secretion of exportable proteins by the exocrine pancreas. Am J Physiol. 1980 Jun;238(6):G467–G477. doi: 10.1152/ajpgi.1980.238.6.G467. [DOI] [PubMed] [Google Scholar]
- Seybold J., Bieger W., Kern H. F. Studies on intracellular transport of secretory proteins in the rat exocrine pancreas. II. Inhibition of antimicrotubular agents. Virchows Arch A Pathol Anat Histol. 1975 Nov 28;368(4):309–327. doi: 10.1007/BF00432309. [DOI] [PubMed] [Google Scholar]
- Stein O., Sanger L., Stein Y. Colchicine-induced inhibition of lipoprotein and protein secretion into the serum and lack of interference with secretion of biliary phospholipids and cholesterol by rat liver in vivo. J Cell Biol. 1974 Jul;62(1):90–103. doi: 10.1083/jcb.62.1.90. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stein O., Stein Y. Colchicine-induced inhibition of very low density lipoprotein release by rat liver in vivo. Biochim Biophys Acta. 1973 Apr 13;306(1):142–147. doi: 10.1016/0005-2760(73)90219-1. [DOI] [PubMed] [Google Scholar]
- Tartakoff A. M., Jamieson J. D., Scheele G. A., Palade G. E. Studies on the pancreas of the guinea pig. Parallel processing and discharge of exocrine proteins. J Biol Chem. 1975 Apr 10;250(7):2671–2677. [PubMed] [Google Scholar]
- Thyberg J., Piasek A., Moskalewski S. Effects of colchicine on the Golgi complex and GERL of cultured rat peritoneal macrophages and epiphyseal chondrocytes. J Cell Sci. 1980 Oct;45:41–58. doi: 10.1242/jcs.45.1.41. [DOI] [PubMed] [Google Scholar]
- Weinstock M., Leblond C. P. Synthesis, migration, and release of precursor collagen by odontoblasts as visualized by radioautography after (3H)proline administration. J Cell Biol. 1974 Jan;60(1):92–127. doi: 10.1083/jcb.60.1.92. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Williams J. A., Lee M. Microtubules and pancreatic amylase release by mouse pancreas in vitro. J Cell Biol. 1976 Dec;71(3):795–806. doi: 10.1083/jcb.71.3.795. [DOI] [PMC free article] [PubMed] [Google Scholar]