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. 1977 Apr 1;73(1):111–127. doi: 10.1083/jcb.73.1.111

Microtubule and microfilament rearrangements during capping of concanavalin A receptors on cultured ovarian granulosa cells

PMCID: PMC2109900  PMID: 558195

Abstract

Thin-section electron microscope analysis of rat and rabbit-cultured granulosa cells treated with concanavalin A (Con A) at 37 degrees C revealed coordinated changes in the cytoplasmic disposition of microfilaments, thick filaments, and microtubules during cap formation and internalization of lectin-receptor complexes. Con A-receptor clustering is accompanied by an accumulation of subplasmalemmal microfilaments which assemble into a loosely woven ring as patches of receptor move centrally on the cell surface. Periodic densities appear in the microfilament ring which becomes reduced in diameter as patches coalesce to form a single central cap. Microtubules and thick filaments emerge associated with the capped membrane. Capping is followed by endocytosis of the con A-receptor complexes. During this process, the microfilament ring is displaced basally into the cytoplasm and endocytic vesicles are transported to the paranuclear Golgi complex along microtubules and thick filaments. Eventually, these vesicles aggregate near the cell center where they are embedded in a dense meshwork of thick filaments. Freeze-fracture analysis of Con A-capped granulosa cells revealed no alteration in the arrangement of peripheral intramembrane particles but large, smooth domains were conspicuous in the capped region of the plasma membrane. The data are discussed with reference to the participation of microtubules and microfilaments in the capping process.

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Selected References

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  1. Albertini D. F., Clark J. I. Membrane-microtubule interactions: concanavalin A capping induced redistribution of cytoplasmic microtubules and colchicine binding proteins. Proc Natl Acad Sci U S A. 1975 Dec;72(12):4976–4980. doi: 10.1073/pnas.72.12.4976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Becker J. S., Oliver J. M., Berlin R. D. Fluorescence techniques for following interactions of microtubule subunits and membranes. Nature. 1975 Mar 13;254(5496):152–154. doi: 10.1038/254152a0. [DOI] [PubMed] [Google Scholar]
  3. Berlin R. D., Oliver J. M., Ukena T. E., Yin H. H. Control of cell surface topography. Nature. 1974 Jan 4;247(5435):45–46. doi: 10.1038/247045a0. [DOI] [PubMed] [Google Scholar]
  4. Bhattacharyya B., Volff J. Membrane-bound tubulin in brain and thyroid tissue. J Biol Chem. 1975 Oct 10;250(19):7639–7646. [PubMed] [Google Scholar]
  5. Brown S. S., Revel J. P. Reversibility of cell surface label rearrangement. J Cell Biol. 1976 Mar;68(3):629–641. doi: 10.1083/jcb.68.3.629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Edelman G. M. Surface modulation in cell recognition and cell growth. Science. 1976 Apr 16;192(4236):218–226. doi: 10.1126/science.769162. [DOI] [PubMed] [Google Scholar]
  7. Edelman G. M., Yahara I., Wang J. L. Receptor mobility and receptor-cytoplasmic interactions in lymphocytes. Proc Natl Acad Sci U S A. 1973 May;70(5):1442–1446. doi: 10.1073/pnas.70.5.1442. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Erickson G. F., Challis J. R., Ryan K. J. A developmental study on the capacity of rabbit granulosa cells to respond to trophic hormones and secrete progesterone in vitro. Dev Biol. 1974 Oct;40(2):208–224. doi: 10.1016/0012-1606(74)90124-9. [DOI] [PubMed] [Google Scholar]
  9. Gruenstein E., Rich A., Weihing R. R. Actin associated with membranes from 3T3 mouse fibroblast and HeLa cells. J Cell Biol. 1975 Jan;64(1):223–234. doi: 10.1083/jcb.64.1.223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hoffstein S., Soberman R., Goldstein I., Weissmann G. Concanavalin A induces microtubule assembly and specific granule discharge in human polymorphonuclear leukocytes. J Cell Biol. 1976 Mar;68(3):781–787. doi: 10.1083/jcb.68.3.781. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ishikawa H., Bischoff R., Holtzer H. Formation of arrowhead complexes with heavy meromyosin in a variety of cell types. J Cell Biol. 1969 Nov;43(2):312–328. [PMC free article] [PubMed] [Google Scholar]
  12. Karnovsky M. J., Unanue E. R. Mapping and migration of lymphocyte surface macromolecules. Fed Proc. 1973 Jan;32(1):55–59. [PubMed] [Google Scholar]
  13. Nicolson G. L. Transmembrane control of the receptors on normal and tumor cells. I. Cytoplasmic influence over surface components. Biochim Biophys Acta. 1976 Apr 13;457(1):57–108. doi: 10.1016/0304-4157(76)90014-9. [DOI] [PubMed] [Google Scholar]
  14. Oliver J. M., Ukena T. E., Berlin R. D. Effects of phagocytosis and colchicine on the distribution of lectin-binding sites on cell surfaces. Proc Natl Acad Sci U S A. 1974 Feb;71(2):394–398. doi: 10.1073/pnas.71.2.394. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Oliver J. M., Zurier R. B., Berlin R. D. Concanavalin a cap formation on polymorphonuclear leukocytes of normal and beige (chediak-higashi) mice. Nature. 1975 Feb 6;253(5491):471–473. doi: 10.1038/253471a0. [DOI] [PubMed] [Google Scholar]
  16. Pollard T. D., Korn E. D. Electron microscopic identification of actin associated with isolated amoeba plasma membranes. J Biol Chem. 1973 Jan 25;248(2):448–450. [PubMed] [Google Scholar]
  17. Pollard T. D., Weihing R. R. Actin and myosin and cell movement. CRC Crit Rev Biochem. 1974 Jan;2(1):1–65. doi: 10.3109/10409237409105443. [DOI] [PubMed] [Google Scholar]
  18. Ryan G. B., Borysenko J. Z., Karnovsky M. J. Factors affecting the redistribution of surface-bound concanavalin A on human polymorphonuclear leukocytes. J Cell Biol. 1974 Aug;62(2):351–365. doi: 10.1083/jcb.62.2.351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Schreiner G. F., Unanue E. R. Calcium-sensitive modulation of Ig capping: evidence supporting a cytoplasmic control of ligand-receptor complexes. J Exp Med. 1976 Jan 1;143(1):15–31. doi: 10.1084/jem.143.1.15. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Silva P. P., Martínez-Palomo A., Gonzalez-Robles A. Membrane structure and surface coat of Entamoeba histolytica. Topochemistry and dynamics of the cell surface: cap formation and microexudate. J Cell Biol. 1975 Mar;64(3):538–550. doi: 10.1083/jcb.64.3.538. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Singer S. J., Nicolson G. L. The fluid mosaic model of the structure of cell membranes. Science. 1972 Feb 18;175(4023):720–731. doi: 10.1126/science.175.4023.720. [DOI] [PubMed] [Google Scholar]
  22. Smith S. B., Revel J. P. Mapping of concanavalin A binding sites on the surface of several cell types. Dev Biol. 1972 Mar;27(3):434–441. doi: 10.1016/0012-1606(72)90183-2. [DOI] [PubMed] [Google Scholar]
  23. Storrie B. Antagonism by dibutyryl adenosine cyclic 3',5'-monophosphate and testololactone of concanavalin A capping. J Cell Biol. 1975 Aug;66(2):392–403. doi: 10.1083/jcb.66.2.392. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Unanue E. R., Karnovsky M. J. Ligand-induced movement of lymphocyte membrane macromolecules. V. Capping, cell movement, and microtubular function in normal and lectin-treated lymphocytes. J Exp Med. 1974 Nov 1;140(5):1207–1220. doi: 10.1084/jem.140.5.1207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. WATSON M. L. Staining of tissue sections for electron microscopy with heavy metals. J Biophys Biochem Cytol. 1958 Jul 25;4(4):475–478. doi: 10.1083/jcb.4.4.475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Weller N. K. Visualization of concanavalin A-binding sites with scanning electron microscopy. J Cell Biol. 1974 Nov;63(2 Pt 1):699–707. doi: 10.1083/jcb.63.2.699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Yahara I., Edelman G. M. Electron microscopic analysis of the modulation of lymphocyte receptor mobility. Exp Cell Res. 1975 Mar 1;91(1):125–142. doi: 10.1016/0014-4827(75)90150-0. [DOI] [PubMed] [Google Scholar]
  28. Yahara I., Edelman G. M. The effects of concanavalin A on the mobility of lymphocyte surface receptors. Exp Cell Res. 1973 Sep;81(1):143–155. doi: 10.1016/0014-4827(73)90121-3. [DOI] [PubMed] [Google Scholar]
  29. de Petris S. Concanavalin A receptors, immunoglobulins, and theta antigen of the lymphocyte surface. Interactions with concanavalin A and with Cytoplasmic structures. J Cell Biol. 1975 Apr;65(1):123–146. doi: 10.1083/jcb.65.1.123. [DOI] [PMC free article] [PubMed] [Google Scholar]

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