Summary
Stimulation of secretion by pilocarpine results in a 70% loss of zymogen granules from pancreatic acinar cell during the first hr after injection of the drug. In previous work (Geuze and Poort, 1973), we found that the amount of membrane stored in the surface of the microvilli and of the numerous infoldings present in highly stimulated cells, increases during the first 2 hr and then decreases again during the 3rd hr after stimulation, concurrently with maximal endocytosis of sorbitol-[su14C].
Further observations on the fine structure of stimulated cells at various time intervals after injection of pilocarpine showed that during the first hr numerous smooth vesicles and multivesicular bodies (mvb's) appear in the apical cytoplasm, while the number of coated vesicles and their relative total volume increase significantly 3 hr after stimulation.
By infusion of ferritin in the pancreatic duct system in vivo and application of cytochemical techniques (osmium impregnation, electron microscope autoradiography and acid phosphatase cytochemistry) it could be established that after stimulated exocytotic secretion, redundant apical cell membrane is withdrawn by at least two routes: 1) During the initial rapid increase of the amount of apical cell membrane, withdrawal is accomplished by interiorization of luminal invaginations into smooth endocytotic vesicles, which in turn give rise to mvb's by infolding and subsequent fission of their limiting membrane. 2) Once the bulk of stored secretion granules has been discharged, endocytotic coated vesicles become gradually more prominent as carriers for redundant cell membrane. The contents of endocytotic structures ultimately become incorporated in residual bodies, suggesting lysosomal degradation of cell membrane prior to eventual reutilization.
Coated vesicles also originate by pinching off from mature Golgi cisternae and condensing vacuoles. A possible function of the coated membranes in the concentration of exportable protein within forming secretory granules is discussed.
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References
Abrahams, S. J., Joltzman, E.: Secretion and endocytosis in insulin-stimulated rat adrenal medulla cells. J. Cell Biol. 56, 540–558 (1973)
Amsterdam, A., Ohad, I., Schramm, M.: Dynamic changes in the ultrastructure of the acinar cell of the rat parotid gland during the secretory cycle. J. Cell Biol. 41, 753–773 (1969)
Arstila, A. U., Jauregui, H. O., Chang, J., Trump, B. F.: Studies on cellular autophagocytosis. Lab. Invest. 24, 162–177 (1971)
Cardell, R. R., Jr., Badenhausen, S., Porter, K. R.: Intestinal triglyceride absorption in the rat. J. Cell Biol. 34, 123–155 (1967)
Douglas, W. W., Nagasawa, J.: Membrane vesiculation at sites of exocytosis in the neurohypophysis, adenohypophysis and adrenal medulla: a device for membrane conservation. J. Physiol. (Lond.) 218, 948 (1971)
Farquhar, M. G., Bainton, D. F., Baggiolini, M., de Duve, C.: Cytochemical localization of acid phosphatase activity in granule fractions from rabbit polymorphonuclear leukocytes. J. Cell Biol. 54, 141–156 (1972)
Farquhar, M. G., Palade, G. E.: Functional evidence for the existence of a third cell type in the renal glomerulus. J. Cell Biol. 13, 55–87 (1962)
Friend, D. S.: Cytochemical staining of multivesicular body and Golgi vesicles. J. Cell Biol. 41, 269–279 (1969)
Friend, D. S., Farquhar, M. G.: Functions of coated vesicles during protein absorption in the rat vas deferens. J. Cell Biol. 35, 357–376 (1967)
Geuze, J. J., Poort, C.: Cell membrane resorption in the rat exocrine pancreas cell after in vivo stimulation of the secretion, as studied by in vitro incubation with extracellular space markers. J. Cell Biol. 57, 159–174 (1973)
Gomori, G.: In: Microscopic Histochemistry, Principles and Practice. University of Chicago Press, p. 189 (1952)
Grynszpan-Winograd, O.: Morphological aspects of exocytosis in the adrenal medulla. Phil. Trans. Soc. Roy. Lond. Ser. B, Biol. Sci. 261, 291–292 (1971)
Hand, A. R..: The fine structure of von Ebner's gland of the rat. J. Cell Biol. 44, 340–353 (1970)
Hand, A. R..: Morphology and cytochemistry of the Golgi apparatus of rat salivary gland acinar cells. Am. J. Anat. 130, 141–157 (1971)
Hermodson, L. H.: Changes in the ultrastructure of the exocrine pancreas of the cat after stimulation of secretin and pancreozymin. In: Electron Microscopy, New York, Academic Press, Inc., yy-11 (1962)
Heuser, J., Reese, T. S..: Evidence for recycling of synaptic vesicle membrane during transmitter release at the frog neuromuscular junction. J. Cell Biol. 57, 315–344 (1973)
Hirsch, J. G., Fedorko, M. E., Cohn, Z. A.: Vesicle fusion and formation at the surface of pinocytotic vacuoles in macrophages. J. Cell Biol. 38, 629–632 (1968)
Holtzman, E.: Stimulation-dependent alterations in peroxidase uptake at lobster neuromuscular junctions. Science 173, 733–736 (1971)
Ichikawa, A.: Fine structural changes in response to hormonal stimulation of the perfused canine pancreas. J. Cell Biol. 24, 369–385 (1965)
Jamieson, J. D., Palade, G. E.: Intracellular transport of secretory proteins in the pancreatic exocrine cell, I. J. Cell Biol. 34, 577–596 (1967)
Jamieson, J. D., Palade, G. E.: Condensing vacuole conversion and zymogen granule discharge in pancreatic exocrine cells: metabolic studies. J. Cell Biol. 48, 503–522 (1971)
Jamieson, J. D., Palade, G. E..: Synthesis, intracellular transport, and discharge of secretory proteins in stimulated pancreatic exocrine cells. J. Cell Biol. 50, 135–158 (1971)
Kanaseki, T., Radota, K.: The “vesicle in the basket”. J. Cell Biol. 42, 202–220 (1969)
Kilarski, W., Jasinski, A.: The formation of multivesicular bodies from the nuclear envelope. J. Cell Biol. 45, 205–211 (1970)
Kopriwa, B. M.: A semiautomatic instrument for the radioautographic coating technique. J. Histochem. Cytochem. 14, 923 (1966)
Kramer, M. F., Geuze, J. J.: Redundant cell membrane regulation in the exocrine pancreas cells after pilocarpine stimulation of the secretion in vivo. In: Advances in Cytopharmacology, vol. 2, p. 87–97. New York: Raven Press (1974)
Lettré, H., Paweletz, N.: Probleme der elektronenmikroskopischen Autoradiographie. Naturwissenschaften 53, 268 (1966)
Locke, M., Collins, J. V.: Protein uptake in multivesicular bodies in the molt-intermolt cycle of an insect. Science 155, 467–469 (1967)
Locke, M., Collins, J. V.: Protein uptake into multivesicular bodies and storage granules in the fat body of an insect. J. Cell Biol. 36, 453–483 (1968)
Masur, S. K., Holtzman, E., Schwartz, I. L., Walter, R.: Correlation between pinocytosis and hydroosmosis induced by neurohypophyseal hormones and mediated by adenosine 3′, 5′-cyclic monophosphate. J. Cell Biol. 49, 582–594 (1971)
Masur, S. K., Holtzman, E., Walter, R.: Hormone-stimulated exocytosis in the toad urinary bladder. J. Cell Biol. 52, 211–219 (1972)
Mata, L. R., Ferreira, J.F.D.: Transport of exogenous peroxidase to Golgi cisternae in the hamster seminal vesicle. J. Microscopie 17, 103–106 (1973)
Matter, A., Orci, L., Forssmann, W. G., Rouiller, Ch.: The stereological analysis of the fine structure of the “micropinocytosis vermiformis” in Kupffer cells of the rat. J. Ultrastruct. Res. 23, 272–279 (1968)
Maul, G. C., Brumbaugh, J. A.: On the possible function of coated vesicles in melanogenesis of the regenerating fowl feather. J. Cell Biol. 48, 41–48 (1971)
Merker, H. J.: Über das Vorkommen multivesiculärer Einschlußkörper im Vaginalepithel der Ratte. Z. Zellforsch. 68, 618–630 (1965)
Nevalainen, T. J.: Effects of pilocarpine stimulation on rat pancreatic acinar cells. Acta path. microbiol. scand. A, Suppl. 210 (1970)
Nicander, L.: A study of multivesicular bodies in the epididymis. J. Ultrastruct. Res. 14, 424–425 (1969)
Novikoff, A. B.: Lysosomes in the physiology and pathology of cells: contributions of staining methods. Ciba Found. Symp. Lysosomes 36 (1963)
Okano, Y., Nakamura, T.: Electron microscopic observations on exocrine pancreas cells in mice following the injection of pilocarpine. J. Electron Micr. (Tokyo) 14, 242–243 (1965)
Palade, G. E.: In: Subcellular particles, T. Hayashi, ed., 64. New York: The Ronald Press Company 1959
Palade, G. E., Bruns, R. R.: Structural modulations of plasmalemmal vesicles. J. Cell Biol. 37, 633–649 (1968)
Rosenbluth, J., Wissig, S. L.: The distribution of exogenous ferritin in toad spinal ganglia and the mechanism of its uptake by neurons. J. Cell Biol. 23, 307–325 (1964)
Roth, T. F., Porter, K. R.: Specialized sites on the cell surface for protein uptake. In: Electron microscopy: Fifth International Congress for Electron Microscopy, Philadelphia, 1962, S. S. Breese, Jr. ed., vol. 2, p. 144. New York: Academic Press Inc. 1962
Shirama, T., Cohen, A. S.: The association of hemidesmosome-like plaque and dense coating with the pinocytotic uptake of a heterologous fibrillar protein (amyloid) by macrophages. J. Ultrastruct. Res. 33, 587–597 (1970)
Simionescu, N., Simionescu, M., Palade, G. E.: Permeability of muscle capillaries to exogenous myoglobin. J. Cell Biol. 57, 424–452 (1973)
Vrensen, G.F.J.M.: Some new aspects of efficiency of electron-microscopic autoradiographs with tritium. J. Histochem. Cytochem. 18, 278–290 (1970)
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Geuze, J.J., Kramer, M.F. Function of coated membranes and multivesicular bodies during membrane regulation in stimulated exocrine pancreas cells. Cell Tissue Res. 156, 1–20 (1974). https://doi.org/10.1007/BF00220098
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DOI: https://doi.org/10.1007/BF00220098