Summary
Using an in situ tannic acid perfusion technique, this study presents evidence that the removal of membrane components from the rat atrial secretory granule membrane after granule exocytosis is mediated by coated vesicles. When tannic acid is used to arrest the post-fusion stages of granule release, coated pit formation occurs on granule membrane, which, although continuous with the sarcolemma, is easily recognised by the membrane omega profile and the continued presence of the granule core. Tannic acid perfusion, before aldehyde fixation, allows a degree of continued cell function, and granule fusions can persist after tannic acid has reached the cell. This results in an increase in the numbers of fusion profiles and the appearance of coated pits on granule membrane at these sites. The proportion of granules with coats increases with perfusion time, suggesting that endocytotic, as well exocytotic events, may be arrested by the action of tannic acid. Coated vesicles are also involved at earlier stages of the release pathway. In other types of secretory system this is considered to represent recycling of membrane proteins as part of the maturation process of the granule. Although arrested granules exhibiting this clathrin coat could have had the coat prior to fusion, as part of the maturation process, our results show that it is more likely to represent a second stage of membrane protein recycling; the postfusion reclamation of proteins from the sarcolemma. This facet of the tannic acid perfusion procedure suggests a general method for quantifying coated pit formation during secretory granule release.
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References
Brooks JC, Carmichael SW (1987) Ultrastructural demonstration of exocytosis in intact and saponin-permeabilized cultured bovine chromaffin cells. Am J Anat 178:85–89
Buma P, Roubos EW, Buijs RM (1984) Ultrastructural demonstration of exocytosis of neural, neuroendocrinal, and endocrinal secretions with an in vitro tannic acid (TARI) method. Histochemistry 80:247–256
Cantin M, Gutkowska J, Thibault G, Milne RW, Ledoux S, Min Li S, Chapeau C, Garcia R, Hamet P, Genest J (1984) Immunocytochemical localization of atrial natriuretic factor in the heart and salivary glands. Histochemistry 80:113–127
Farquhar MG (1983) Multiple pathways of exocytosis, endocytosis and membrane recycling: validation of a Golgi route. Fed Proc 42:2407–2413
Goldstein JL, Brown MS, Anderson RB, Russel DW, Scheider WJ (1985) Receptor-mediated endocytosis: concepts emerging from the LDL receptor system. Annu Rev Cell Biol 1:1–39
Herzog V (1981) Pathways of endocytosis in secretory cells. Trends Biochem Sci 6:319–322
Herzog V, Farquhar MG (1977) Luminal membrane retrieved after exocytosis reaches most Golgi cisternae in secretory cells. Proc Natl Acad Sci USA 74:5073–5077
Heuser J (1989) The role of coated vesicles in recycling of synaptic vesicle membrane. Cell Biol Int Reps 13:1963–1976
Kurihara H, Kitajima K, Senda T, Fujita H, Nakajima (1986) Multigranular exocytosis induced by phospholipase A2-activators, melittin and mastoparan, in rat anterior pituitary cells. Cell Tissue Res 243:311–316
Miller TM, Heuser JE (1984) Endocytosis of synaptic vesicle membrane at the frog neuromuscular junction. J Cell Biol 98:685–698
Morris JF, Pow DV (1988) Capturing and quantifying the exocytotic event. J Exp Biol 139:81–103
Morris JF, Pow DV, Shaw F (1988) Release of neuropeptides from manocellular neurones: Does anatomical compartmentation have a functional significance? In: Pickering BT, Wakerley JB, Summerlee AJS (eds) Neurosecretion. Cellular aspects of the production and release of neuropeptides. Plenum, New York, pp 113–122
Newman TM, Severs NJ (1990a) Arrested exocytosis of atrial secretory granules. J Mol Cell Cardiol 22:771–786
Newman TM, Severs NJ (1990b) Arrested exocytosis of synaptic vesicles at a glutamatergic synapse, the insect fast excitatory neuromuscular junction. Cell Biol Int Rept 14:849–863
Orci L, Ravazzola M, Amherdt M, Louvard D, Perrelet (1985) Clathrin-immunoreactive sites in the Golgi apparatus are concentrated at the trans pole in polypeptide hormone-secreting cells. Proc Natl Acad Sci USA 82:5385–5389
Ottosen PD, Courtoy PJ, Farquhar MG (1980) Pathways followed by membrane recovered from the surface of plasma cells and myeloma cells. J Exp Med 152:1–19
Patzak A, Winkler H (1986) Exocytotic exposure and recycling of membrane antigens of chromaffin granules. J Cell Biol 102:510–515
Patzak A, Aunis D, Lanley K (1987) Membrane recycling after exocytosis: an ultrastructural study of cultured chromaffin cells. Exp Cell Res 171:346–356
Pearse BMF, Robinson MS (1990) Clathrin, adapters, and sorting. Annu Rev Cell Biol 6:151–171
Pow DV, Morris JF (1990) Exocytotic release of oxytocin and vasopressin from dendrites in the rat supraoptic nucleus. J Neuroendocrinol [Suppl] 1:50–52
Pow DV, Morris JF (1991) Membrane routing during exocytosis and endocytosis in neuroendocrine neurones and endocrine cells: use of colloidal gold particles and immunocytochemical discrimination of membrane compartments. Cell Tissue Res 264:299–316
Rothman JE, Pettegrew HG, Fine RE (1980) Transport of the membrane protein of the vesicular stomatitis virus to the cell surface in two stages by clathrin-coated vesicles. J Cell Biol 86:162–171
Severs NJ (1989a) Constituent cells of the heart and isolated cell models in cardiovascular research. In: Piper HM, Isenberg G (eds) Isolated adult cardiomyocytes. CRC Press, Boca Raton, pp 3–41
Severs NJ (1989b) Membrane cytochemistry of the atrial peptide secretory pathway. Am J Physiol Heart Circ Physiol 257:H1587-H1598
Steinman RM, Mellman IS, Muller WA, Cohn ZA (1983) Endocytosis and the recycling of membrane. J Cell Biol 96:1–27
Theron JJ, Biagio R, Meyer AC, Boekkooi S (1978) Ultrastructural observations on the maturation of granules in atrial myocardium. J Mol Cell Cardiol 10:567–572
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Newman, T.M., Severs, N.J. Coated vesicles are implicated in the post-fusion retrieval of the membrane of rat atrial secretory granules. Cell Tissue Res 268, 463–469 (1992). https://doi.org/10.1007/BF00319153
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DOI: https://doi.org/10.1007/BF00319153