Abstract
The potential involvement of actin and fodrin (brain spectrin) in secretory events has been assessed in primary cultured guinea pig parotid acinar cells, using as a tool affinity purified anti-alpha- fodrin antibody, phalloidin, and immunofluorescence techniques. In resting parotid acinar cells fodrin and actin appeared as a continuous ring under the plasma membrane of most of the cells. Upon stimulation with secretagogues fodrin and actin labeling at the level of the plasma membrane disappeared almost completely. To establish a correlation between secretion and cytoskeletal changes at the individual cell level, anti-alpha-amylase-antibodies were used to label secreted amylase exposed at the surface of secreting cells. The number of cells expressing alpha-amylase on their surface followed bulk secretion of alpha-amylase. A strict correlation between secretion and alteration of the actin-fodrin labeling was observed at the individual cell level. The cytoskeletal changes occurred in parallel with secretion independently of the secretagogue used (carbamoylcholine in the presence of Ca2+, isoproterenol in presence or absence of Ca2+, forskolin, or dibutyryl-cyclic-AMP). The changes were reversible upon removal of the secretagogue. Since Ca2+, as well as cAMP-mediated secretion, was associated with the same kind of cytoskeletal changes, a reorganization of the cytoskeleton may play an essential part in regulated secretion.
Full Text
The Full Text of this article is available as a PDF (965.5 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Backman L. Protein cytoskeleton. Functional or futile phosphorus? Nature. 1988 Aug 25;334(6184):653–654. doi: 10.1038/334653a0. [DOI] [PubMed] [Google Scholar]
- Burgoyne R. D., Morgan A., O'Sullivan A. J. The control of cytoskeletal actin and exocytosis in intact and permeabilized adrenal chromaffin cells: role of calcium and protein kinase C. Cell Signal. 1989;1(4):323–334. doi: 10.1016/0898-6568(89)90051-x. [DOI] [PubMed] [Google Scholar]
- Burridge K., Kelly T., Mangeat P. Nonerythrocyte spectrins: actin-membrane attachment proteins occurring in many cell types. J Cell Biol. 1982 Nov;95(2 Pt 1):478–486. doi: 10.1083/jcb.95.2.478. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Butcher F. R., Putney J. W., Jr Regulation of parotid gland function by cyclic nucleotides and calcium. Adv Cyclic Nucleotide Res. 1980;13:215–249. [PubMed] [Google Scholar]
- Domenech C., Machado-De Domenech E., Söling H. D. Regulation of acetyl-CoA:1-alkyl-sn-glycero-3-phosphocholine O2-acetyltransferase (lyso-PAF-acetyltransferase) in exocrine glands. Evidence for an activation via phosphorylation by calcium/calmodulin-dependent protein kinase. J Biol Chem. 1987 Apr 25;262(12):5671–5676. [PubMed] [Google Scholar]
- Fujimoto T., Ogawa K. Retrieving vesicles in secretion-induced rat chromaffin cells contain fodrin. J Histochem Cytochem. 1989 Nov;37(11):1589–1599. doi: 10.1177/37.11.2509551. [DOI] [PubMed] [Google Scholar]
- Glenney J. R., Jr, Glenney P., Weber K. F-actin-binding and cross-linking properties of porcine brain fodrin, a spectrin-related molecule. J Biol Chem. 1982 Aug 25;257(16):9781–9787. [PubMed] [Google Scholar]
- Goodman S. R., Krebs K. E., Whitfield C. F., Riederer B. M., Zagon I. S. Spectrin and related molecules. CRC Crit Rev Biochem. 1988;23(2):171–234. doi: 10.3109/10409238809088319. [DOI] [PubMed] [Google Scholar]
- Lamb N. J., Fernandez A., Conti M. A., Adelstein R., Glass D. B., Welch W. J., Feramisco J. R. Regulation of actin microfilament integrity in living nonmuscle cells by the cAMP-dependent protein kinase and the myosin light chain kinase. J Cell Biol. 1988 Jun;106(6):1955–1971. doi: 10.1083/jcb.106.6.1955. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Levine J., Willard M. Fodrin: axonally transported polypeptides associated with the internal periphery of many cells. J Cell Biol. 1981 Sep;90(3):631–642. doi: 10.1083/jcb.90.3.631. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nargessi R. D., Landon J. Indirect quenching fluoroimmunoassay. Methods Enzymol. 1981;74(Pt 100):60–79. doi: 10.1016/0076-6879(81)74005-9. [DOI] [PubMed] [Google Scholar]
- Perrin D., Aunis D. Reorganization of alpha-fodrin induced by stimulation in secretory cells. Nature. 1985 Jun 13;315(6020):589–592. doi: 10.1038/315589a0. [DOI] [PubMed] [Google Scholar]
- Perrin D., Langley O. K., Aunis D. Anti-alpha-fodrin inhibits secretion from permeabilized chromaffin cells. Nature. 1987 Apr 2;326(6112):498–501. doi: 10.1038/326498a0. [DOI] [PubMed] [Google Scholar]
- Söling H. D., Fest W., Schmidt T., Esselmann H., Bachmann V. Signal transmission in exocrine cells is associated with rapid activity changes of acyltransferases and diacylglycerol kinase due to reversible protein phosphorylation. J Biol Chem. 1989 Jun 25;264(18):10643–10648. [PubMed] [Google Scholar]
- Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tsien R. Y. New calcium indicators and buffers with high selectivity against magnesium and protons: design, synthesis, and properties of prototype structures. Biochemistry. 1980 May 27;19(11):2396–2404. doi: 10.1021/bi00552a018. [DOI] [PubMed] [Google Scholar]