Abstract
The physiology of protein intracellular transport and secretion by cell types thought to be free from short-term control has been compared with that of the pancreatic acinar cell, using pulse-chase protocols to follow biosynthetically-labeled secretory products. Data previously obtained (Tartakoff, A.M., and P. Vassalli. J. Exp. Med. 146:1332-1345) has shown that plasma-cell immunoglobulin (Ig) secretion is inhibited by respiratory inhibitors, by partial Na/K equilibration effected by the carboxylic ionophore monensin, and by calcium withdrawal effected by the carboxylic ionophore A 23187 in the presence of ethylene glycol bis (beta-aminoethylether)-N,N,N',N'-tetraacetic acid (EGTA) and absence of calcium. We report here that both inhibition of respiration and treatment with monensin slow secretion by fibroblasts, and also macrophages and slow intracellular transport (though not discharge per se) by the exocrine pancreatic cells. Attempted calcium withdrawal is inhibitory for fibroblasts but not for macrophages. The elimination of extracellular calcium or addition of 50 mM KCl has no major effect on secretory rate of either fibroblasts or macrophages. Electron microscopic examination of all cell types shows that monensin causes a rapid and impressive dilation of Golgi elements. Combined cell fractionation and autoradiographic studies of the pancreas show that the effect of monensin is exerted at the point of the exit of secretory protein from the Golgi apparatus. Other steps in intracellular transport proceed at normal rates. These observations suggest a common effect of the cytoplasmic Na/K balance at the Golgi level and lead to a model of intracellular transport in which secretory product obligatorily passes through Golgi elements (cisternae?) that are sensitive to monensin. Thus, intracellular transport follows a similar course in both regulated and nonregulated secretory cells up to the level of distal Golgi elements.
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- Batzri S., Selinger Z., Schramm M., Robinovitch M. R. Potassium release mediated by the epinephrine -receptor in rat parotid slices. Properties and relation to enzyme secretion. J Biol Chem. 1973 Jan 10;248(1):361–368. [PubMed] [Google Scholar]
- Bergeron J. J., Ehrenreich J. H., Siekevitz P., Palade G. E. Golgi fractions prepared from rat liver homogenates. II. Biochemical characterization. J Cell Biol. 1973 Oct;59(1):73–88. doi: 10.1083/jcb.59.1.73. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bieger W., Seybold J., Kern H. F. Studies on intracellular transport of secretory proteins in the rat exocrine pancreas. III. Effect of cobalt, lanthanum and antimycin A. Virchows Arch A Pathol Anat Histol. 1975 Nov 28;368(4):329–345. doi: 10.1007/BF00432310. [DOI] [PubMed] [Google Scholar]
- COHN Z. A., BENSON B. THE DIFFERENTIATION OF MONONUCLEAR PHAGOCYTES. MORPHOLOGY, CYTOCHEMISTRY, AND BIOCHEMISTRY. J Exp Med. 1965 Jan 1;121:153–170. doi: 10.1084/jem.121.1.153. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ehrenreich J. H., Bergeron J. J., Siekevitz P., Palade G. E. Golgi fractions prepared from rat liver homogenates. I. Isolation procedure and morphological characterization. J Cell Biol. 1973 Oct;59(1):45–72. doi: 10.1083/jcb.59.1.45. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Eimerl S., Savion N., Heichal O., Selinger Z. Induction of enzyme secretion in rat pancreatic slices using the ionophore A-23187 and calcium. An experimental bypass of the hormone receptor pathway. J Biol Chem. 1974 Jun 25;249(12):3991–3993. [PubMed] [Google Scholar]
- Gordon S., Todd J., Cohn Z. A. In vitro synthesis and secretion of lysozyme by mononuclear phagocytes. J Exp Med. 1974 May 1;139(5):1228–1248. doi: 10.1084/jem.139.5.1228. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hand A. R., Oliver C. Relationship between the Golgi apparatus, GERL, and secretory granules in acinar cells of the rat exorbital lacrimal gland. J Cell Biol. 1977 Aug;74(2):399–413. doi: 10.1083/jcb.74.2.399. [DOI] [PMC free article] [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]
- Jamieson J. D., Palade G. E. Intracellular transport of secretory proteins in the pancreatic exocrine cell. IV. Metabolic requirements. J Cell Biol. 1968 Dec;39(3):589–603. doi: 10.1083/jcb.39.3.589. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kraehenbuhl J. P., Racine L., Jamieson J. D. Immunocytochemical localization of secretory proteins in bovine pancreatic exocrine cells. J Cell Biol. 1977 Feb;72(2):406–423. doi: 10.1083/jcb.72.2.406. [DOI] [PMC free article] [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]
- Leduc E. H., Avrameas S., Bouteille M. Ultrastructural localization of antibody in differentiating plasma cells. J Exp Med. 1968 Jan 1;127(1):109–118. doi: 10.1084/jem.127.1.109. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Meldolesi J., Cova D. Composition of cellular membranes in the pancreas of the guinea pig. IV. Polyacrylamide gel electrophoresis and amino acid composition of membrane proteins. J Cell Biol. 1972 Oct;55(1):1–18. doi: 10.1083/jcb.55.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Meldolesi J., Jamieson J. D., Palade G. E. Composition of cellular membranes in the pancreas of the guinea pig. II. Lipids. J Cell Biol. 1971 Apr;49(1):130–149. doi: 10.1083/jcb.49.1.130. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Novikoff A. B., Mori M., Quintana N., Yam A. Studies of the secretory process in the mammalian exocrine pancreas. I. The condensing vacuoles. J Cell Biol. 1977 Oct;75(1):148–165. doi: 10.1083/jcb.75.1.148. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Olsen B. R., Hoffmann H., Prockop D. J. Interchain disulfide bonds at the COOH-terminal end of procollagen synthesized by matrix-free cells from chick embryonic tendon and cartilage. Arch Biochem Biophys. 1976 Jul;175(1):341–350. doi: 10.1016/0003-9861(76)90516-6. [DOI] [PubMed] [Google Scholar]
- Scheele G. A., Palade G. E. Studies on the guinea pig pancreas. Parallel discharge of exocrine enzyme activities. J Biol Chem. 1975 Apr 10;250(7):2660–2670. [PubMed] [Google Scholar]
- Scheele G. A., Palade G. E., Tartakoff A. M. Cell fractionation studies on the guinea pig pancreas. Redistribution of exocrine proteins during tissue homogenization. J Cell Biol. 1978 Jul;78(1):110–130. doi: 10.1083/jcb.78.1.110. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tartakoff A. M., Jamieson J. D. Subcellular fractionation of the pancreas. Methods Enzymol. 1974;31:41–59. doi: 10.1016/0076-6879(74)31006-3. [DOI] [PubMed] [Google Scholar]
- Tartakoff A. M., Vassalli P. Plasma cell immunoglobulin secretion: arrest is accompanied by alterations of the golgi complex. J Exp Med. 1977 Nov 1;146(5):1332–1345. doi: 10.1084/jem.146.5.1332. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Unkeless J. C., Gordon S., Reich E. Secretion of plasminogen activator by stimulated macrophages. J Exp Med. 1974 Apr 1;139(4):834–850. doi: 10.1084/jem.139.4.834. [DOI] [PMC free article] [PubMed] [Google Scholar]