Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1988 Jan 1;106(1):97–109. doi: 10.1083/jcb.106.1.97

Heterogeneity in lymphocyte spectrin distribution: ultrastructural identification of a new spectrin-rich cytoplasmic structure

PMCID: PMC2114939  PMID: 3276715

Abstract

Spectrin-like proteins are found in a wide variety of non-erythroid cells where they generally occur in the cell cortex near the plasma membrane. To determine the intracellular distribution of alpha-spectrin (alpha-fodrin) in lymphocytes, we have developed an immunoperoxidase method to localize this protein at the ultrastructural level. Of considerable interest, particularly with regard to our efforts to determine the function of spectrin in this cell type, was the finding that its subcellular localization and its relationship with the plasma membrane can vary dramatically. Based on its position in the cell, alpha-spectrin can occur in two forms in lymphocytes: one that associates closely with the plasma membrane and another that occurs at some distance from the cell periphery, either as a single large aggregate or as several smaller ones. The single large aggregate of spectrin is a stable feature in a number of lymphocyte cell lines and hybrids which were used to examine its ultrastructural characteristics. A previously undescribed cellular structure, consisting of a meshwork of spectrin filaments and membranous vesicles, was identified in these cells. This structure could be induced to dissipate in response to membrane perturbants (e.g., hyperthermia and phorbol esters, known effectors of lymphocyte function and differentiation) and the patterns resulting from the redistribution of spectrin were a reflection of those observed routinely in lymphocytes in situ. The correlation between naturally occurring spectrin localization patterns and those seen after membrane perturbation suggested the possibility that spectrin distribution is indicative of particular maturation stages or functional states in lymphocytes. The implications of these findings with regard to the role of spectrin in lymphocyte function are discussed.

Full Text

The Full Text of this article is available as a PDF (7.4 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Apgar J. R., Herrmann S. H., Robinson J. M., Mescher M. F. Triton X-100 extraction of P815 tumor cells: evidence for a plasma membrane skeleton structure. J Cell Biol. 1985 May;100(5):1369–1378. doi: 10.1083/jcb.100.5.1369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bennett V. The membrane skeleton of human erythrocytes and its implications for more complex cells. Annu Rev Biochem. 1985;54:273–304. doi: 10.1146/annurev.bi.54.070185.001421. [DOI] [PubMed] [Google Scholar]
  3. Bodine D. M., 4th, Birkenmeier C. S., Barker J. E. Spectrin deficient inherited hemolytic anemias in the mouse: characterization by spectrin synthesis and mRNA activity in reticulocytes. Cell. 1984 Jul;37(3):721–729. doi: 10.1016/0092-8674(84)90408-2. [DOI] [PubMed] [Google Scholar]
  4. Bourguignon L. Y., Bourguignon G. J. Capping and the cytoskeleton. Int Rev Cytol. 1984;87:195–224. doi: 10.1016/s0074-7696(08)62443-2. [DOI] [PubMed] [Google Scholar]
  5. Bourguignon L. Y., Suchard S. J., Nagpal M. L., Glenney J. R., Jr A T-lymphoma transmembrane glycoprotein (gp180) is linked to the cytoskeletal protein, fodrin. J Cell Biol. 1985 Aug;101(2):477–487. doi: 10.1083/jcb.101.2.477. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cohen C. M. The molecular organization of the red cell membrane skeleton. Semin Hematol. 1983 Jul;20(3):141–158. [PubMed] [Google Scholar]
  7. Elgsaeter A., Stokke B. T., Mikkelsen A., Branton D. The molecular basis of erythrocyte shape. Science. 1986 Dec 5;234(4781):1217–1223. doi: 10.1126/science.3775380. [DOI] [PubMed] [Google Scholar]
  8. Glenney J., Glenney P. Co-expression of spectrin and fodrin in Friend erythroleukemic cells treated with DMSO. Exp Cell Res. 1984 May;152(1):15–21. doi: 10.1016/0014-4827(84)90225-8. [DOI] [PubMed] [Google Scholar]
  9. Goodman S. R., Shiffer K. The spectrin membrane skeleton of normal and abnormal human erythrocytes: a review. Am J Physiol. 1983 Mar;244(3):C121–C141. doi: 10.1152/ajpcell.1983.244.3.C121. [DOI] [PubMed] [Google Scholar]
  10. Hirokawa N., Cheney R. E., Willard M. Location of a protein of the fodrin-spectrin-TW260/240 family in the mouse intestinal brush border. Cell. 1983 Mar;32(3):953–965. doi: 10.1016/0092-8674(83)90080-6. [DOI] [PubMed] [Google Scholar]
  11. Hughes C. S., Repasky E. A., Bankert R. B., Johnson R. J., Subjeck J. R. Effects of hyperthermia on spectrin expression patterns of murine lymphocytes. Radiat Res. 1987 Oct;112(1):116–123. [PubMed] [Google Scholar]
  12. Jampel H. D., Duff G. W., Gershon R. K., Atkins E., Durum S. K. Fever and immunoregulation. III. Hyperthermia augments the primary in vitro humoral immune response. J Exp Med. 1983 Apr 1;157(4):1229–1238. doi: 10.1084/jem.157.4.1229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kwock L., Lin P. S., Hefter K. A comparison of the effects of hyperthermia on cell growth in human T and B lymphoid cells: relationship to alterations in plasma membrane transport properties. Radiat Res. 1985 Jan;101(1):197–206. [PubMed] [Google Scholar]
  14. Lee J. K., Repasky E. A. Cytoskeletal polarity in mammalian lymphocytes in situ. Cell Tissue Res. 1987 Jan;247(1):195–202. doi: 10.1007/BF00216562. [DOI] [PubMed] [Google Scholar]
  15. Levine J., Willard M. Redistribution of fodrin (a component of the cortical cytoplasm) accompanying capping of cell surface molecules. Proc Natl Acad Sci U S A. 1983 Jan;80(1):191–195. doi: 10.1073/pnas.80.1.191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mangeat P. H., Burridge K. Immunoprecipitation of nonerythrocyte spectrin within live cells following microinjection of specific antibodies: relation to cytoskeletal structures. J Cell Biol. 1984 Apr;98(4):1363–1377. doi: 10.1083/jcb.98.4.1363. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Marchesi V. T. Stabilizing infrastructure of cell membranes. Annu Rev Cell Biol. 1985;1:531–561. doi: 10.1146/annurev.cb.01.110185.002531. [DOI] [PubMed] [Google Scholar]
  18. McLean I. W., Nakane P. K. Periodate-lysine-paraformaldehyde fixative. A new fixation for immunoelectron microscopy. J Histochem Cytochem. 1974 Dec;22(12):1077–1083. doi: 10.1177/22.12.1077. [DOI] [PubMed] [Google Scholar]
  19. Nelson W. J., Colaço C. A., Lazarides E. Involvement of spectrin in cell-surface receptor capping in lymphocytes. Proc Natl Acad Sci U S A. 1983 Mar;80(6):1626–1630. doi: 10.1073/pnas.80.6.1626. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Nelson W. J., Veshnock P. J. Dynamics of membrane-skeleton (fodrin) organization during development of polarity in Madin-Darby canine kidney epithelial cells. J Cell Biol. 1986 Nov;103(5):1751–1765. doi: 10.1083/jcb.103.5.1751. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Nicolson G. L., Marchesi V. T., Singer S. J. The localization of spectrin on the inner surface of human red blood cell membranes by ferritin-conjugated antibodies. J Cell Biol. 1971 Oct;51(1):265–272. doi: 10.1083/jcb.51.1.265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Pauly J. L., Bankert R. B., Repasky E. A. Immunofluorescent patterns of spectrin in lymphocyte cell lines. J Immunol. 1986 Jan;136(1):246–253. [PubMed] [Google Scholar]
  23. Pearl M., Fishkind D., Mooseker M., Keene D., Keller T., 3rd Studies on the spectrin-like protein from the intestinal brush border, TW 260/240, and characterization of its interaction with the cytoskeleton and actin. J Cell Biol. 1984 Jan;98(1):66–78. doi: 10.1083/jcb.98.1.66. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. 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]
  26. RODRIGUEZ J., DEINHARDT F. Preparation of a semipermanent mounting medium for fluorescent antibody studies. Virology. 1960 Oct;12:316–317. doi: 10.1016/0042-6822(60)90205-1. [DOI] [PubMed] [Google Scholar]
  27. Repasky E. A., Granger B. L., Lazarides E. Widespread occurrence of avian spectrin in nonerythroid cells. Cell. 1982 Jul;29(3):821–833. doi: 10.1016/0092-8674(82)90444-5. [DOI] [PubMed] [Google Scholar]
  28. Repasky E. A., Symer D. E., Bankert R. B. Spectrin immunofluorescence distinguishes a population of naturally capped lymphocytes in situ. J Cell Biol. 1984 Jul;99(1 Pt 1):350–355. doi: 10.1083/jcb.99.1.350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Tokuyasu K. T., Schekman R., Singer S. J. Domains of receptor mobility and endocytosis in the membranes of neonatal human erythrocytes and reticulocytes are deficient in spectrin. J Cell Biol. 1979 Feb;80(2):481–486. doi: 10.1083/jcb.80.2.481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. White J., Haskins K. M., Marrack P., Kappler J. Use of I region-restricted, antigen-specific T cell hybridomas to produce idiotypically specific anti-receptor antibodies. J Immunol. 1983 Mar;130(3):1033–1037. [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES