Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1994 Mar 2;124(6):1039–1046. doi: 10.1083/jcb.124.6.1039

Expression of a beta 1-related integrin by oligodendroglia in primary culture: evidence for a functional role in myelination

PMCID: PMC2119971  PMID: 7510711

Abstract

We have investigated the expression of integrins by rat oligodendroglia grown in primary culture and the functional role of these proteins in myelinogenesis. Immunochemical analysis, using antibodies to a number of alpha and beta integrin subunits, revealed that oligodendrocytes express only one detectable integrin receptor complex (alpha OL beta OL). This complex is immunoprecipitated by a polyclonal anti-human beta 1 integrin subunit antibody. In contrast, astrocytes, the other major glial cell type in brain, express multiple integrins including alpha 1 beta 1, alpha 3 beta 1, and alpha 5 beta 1 complexes that are immunologically and electrophoretically indistinguishable from integrins expressed by rat fibroblasts. The beta subunit of the oligodendrocyte integrin (beta OL) and rat fibroblast beta 1 have different electrophoretic mobilities in SDS-PAGE. However, the two beta subunits appear to be highly related based on immunological cross- reactivity and one-dimensional peptide mapping. After removal of N- linked carbohydrate chains, beta OL and beta 1 comigrated in SDS-PAGE and peptide maps of the two deglycosylated subunits were identical, suggesting differential glycosylation of beta 1 and beta OL accounts entirely for their size differences. The oligodendrocyte alpha subunit, alpha OL, was not immunoprecipitated by antibodies against well characterized alpha chains which are known to associate with beta 1 (alpha 3, alpha 4, and alpha 5). However, an antibody to alpha 8, a more recently identified integrin subunit, did precipitate two integrin subunits with electrophoretic mobilities in SDS-PAGE identical to alpha OL and beta OL. Functional studies indicated that disruption of oligodendrocyte adhesion to a glial-derived matrix by an RGD-containing synthetic peptide resulted in a substantial decrease in the level of mRNAs for several myelin components including myelin basic protein (MBP), proteolipid protein (PLP), and cyclic nucleotide phosphodiesterase (CNP). These results suggest that integrin-mediated adhesion of oligodendrocytes may trigger signal(s) that induce the expression of myelin genes and thus influence oligodendrocyte differentiation.

Full Text

The Full Text of this article is available as a PDF (1.5 MB).

Selected References

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

  1. Akiyama S. K., Yamada S. S., Yamada K. M. Analysis of the role of glycosylation of the human fibronectin receptor. J Biol Chem. 1989 Oct 25;264(30):18011–18018. [PubMed] [Google Scholar]
  2. Albelda S. M., Buck C. A. Integrins and other cell adhesion molecules. FASEB J. 1990 Aug;4(11):2868–2880. [PubMed] [Google Scholar]
  3. Bossy B., Bossy-Wetzel E., Reichardt L. F. Characterization of the integrin alpha 8 subunit: a new integrin beta 1-associated subunit, which is prominently expressed on axons and on cells in contact with basal laminae in chick embryos. EMBO J. 1991 Sep;10(9):2375–2385. doi: 10.1002/j.1460-2075.1991.tb07776.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cardwell M. C., Rome L. H. Evidence that an RGD-dependent receptor mediates the binding of oligodendrocytes to a novel ligand in a glial-derived matrix. J Cell Biol. 1988 Oct;107(4):1541–1549. doi: 10.1083/jcb.107.4.1541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cardwell M. C., Rome L. H. RGD-containing peptides inhibit the synthesis of myelin-like membrane by cultured oligodendrocytes. J Cell Biol. 1988 Oct;107(4):1551–1559. doi: 10.1083/jcb.107.4.1551. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cheresh D. A., Smith J. W., Cooper H. M., Quaranta V. A novel vitronectin receptor integrin (alpha v beta x) is responsible for distinct adhesive properties of carcinoma cells. Cell. 1989 Apr 7;57(1):59–69. doi: 10.1016/0092-8674(89)90172-4. [DOI] [PubMed] [Google Scholar]
  7. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  8. Cleveland D. W., Fischer S. G., Kirschner M. W., Laemmli U. K. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem. 1977 Feb 10;252(3):1102–1106. [PubMed] [Google Scholar]
  9. Cleveland D. W., Lopata M. A., MacDonald R. J., Cowan N. J., Rutter W. J., Kirschner M. W. Number and evolutionary conservation of alpha- and beta-tubulin and cytoplasmic beta- and gamma-actin genes using specific cloned cDNA probes. Cell. 1980 May;20(1):95–105. doi: 10.1016/0092-8674(80)90238-x. [DOI] [PubMed] [Google Scholar]
  10. Dedhar S., Gray V. Isolation of a novel integrin receptor mediating Arg-Gly-Asp-directed cell adhesion to fibronectin and type I collagen from human neuroblastoma cells. Association of a novel beta 1-related subunit with alpha v. J Cell Biol. 1990 Jun;110(6):2185–2193. doi: 10.1083/jcb.110.6.2185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Diamond M. S., Staunton D. E., Marlin S. D., Springer T. A. Binding of the integrin Mac-1 (CD11b/CD18) to the third immunoglobulin-like domain of ICAM-1 (CD54) and its regulation by glycosylation. Cell. 1991 Jun 14;65(6):961–971. doi: 10.1016/0092-8674(91)90548-d. [DOI] [PubMed] [Google Scholar]
  12. Erle D. J., Rüegg C., Sheppard D., Pytela R. Complete amino acid sequence of an integrin beta subunit (beta 7) identified in leukocytes. J Biol Chem. 1991 Jun 15;266(17):11009–11016. [PubMed] [Google Scholar]
  13. Fischer G., Künemund V., Schachner M. Neurite outgrowth patterns in cerebellar microexplant cultures are affected by antibodies to the cell surface glycoprotein L1. J Neurosci. 1986 Feb;6(2):605–612. doi: 10.1523/JNEUROSCI.06-02-00605.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hemler M. E., Crouse C., Takada Y., Sonnenberg A. Multiple very late antigen (VLA) heterodimers on platelets. Evidence for distinct VLA-2, VLA-5 (fibronectin receptor), and VLA-6 structures. J Biol Chem. 1988 Jun 5;263(16):7660–7665. [PubMed] [Google Scholar]
  15. Hemler M. E., Huang C., Schwarz L. The VLA protein family. Characterization of five distinct cell surface heterodimers each with a common 130,000 molecular weight beta subunit. J Biol Chem. 1987 Mar 5;262(7):3300–3309. [PubMed] [Google Scholar]
  16. Hogervorst F., Kuikman I., von dem Borne A. E., Sonnenberg A. Cloning and sequence analysis of beta-4 cDNA: an integrin subunit that contains a unique 118 kd cytoplasmic domain. EMBO J. 1990 Mar;9(3):765–770. doi: 10.1002/j.1460-2075.1990.tb08171.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hynes R. O. Integrins: a family of cell surface receptors. Cell. 1987 Feb 27;48(4):549–554. doi: 10.1016/0092-8674(87)90233-9. [DOI] [PubMed] [Google Scholar]
  18. Hynes R. O. Integrins: versatility, modulation, and signaling in cell adhesion. Cell. 1992 Apr 3;69(1):11–25. doi: 10.1016/0092-8674(92)90115-s. [DOI] [PubMed] [Google Scholar]
  19. Jaconi M. E., Theler J. M., Schlegel W., Appel R. D., Wright S. D., Lew P. D. Multiple elevations of cytosolic-free Ca2+ in human neutrophils: initiation by adherence receptors of the integrin family. J Cell Biol. 1991 Mar;112(6):1249–1257. doi: 10.1083/jcb.112.6.1249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kajiji S., Tamura R. N., Quaranta V. A novel integrin (alpha E beta 4) from human epithelial cells suggests a fourth family of integrin adhesion receptors. EMBO J. 1989 Mar;8(3):673–680. doi: 10.1002/j.1460-2075.1989.tb03425.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kawano T., Takasaki S., Tao T. W., Kobata A. Altered glycosylation of beta 1 integrins associated with reduced adhesiveness to fibronectin and laminin. Int J Cancer. 1993 Jan 2;53(1):91–96. doi: 10.1002/ijc.2910530118. [DOI] [PubMed] [Google Scholar]
  22. Kornberg L. J., Earp H. S., Turner C. E., Prockop C., Juliano R. L. Signal transduction by integrins: increased protein tyrosine phosphorylation caused by clustering of beta 1 integrins. Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8392–8396. doi: 10.1073/pnas.88.19.8392. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Krissansen G. W., Elliott M. J., Lucas C. M., Stomski F. C., Berndt M. C., Cheresh D. A., Lopez A. F., Burns G. F. Identification of a novel integrin beta subunit expressed on cultured monocytes (macrophages). Evidence that one alpha subunit can associate with multiple beta subunits. J Biol Chem. 1990 Jan 15;265(2):823–830. [PubMed] [Google Scholar]
  24. Malek-Hedayat S., Rome L. H. Expression of multiple integrins and extracellular matrix components by C6 glioma cells. J Neurosci Res. 1992 Mar;31(3):470–478. doi: 10.1002/jnr.490310309. [DOI] [PubMed] [Google Scholar]
  25. Markwell M. A., Fox C. F. Surface-specific iodination of membrane proteins of viruses and eucaryotic cells using 1,3,4,6-tetrachloro-3alpha,6alpha-diphenylglycoluril. Biochemistry. 1978 Oct 31;17(22):4807–4817. doi: 10.1021/bi00615a031. [DOI] [PubMed] [Google Scholar]
  26. Martini R., Schachner M. Immunoelectron microscopic localization of neural cell adhesion molecules (L1, N-CAM, and MAG) and their shared carbohydrate epitope and myelin basic protein in developing sciatic nerve. J Cell Biol. 1986 Dec;103(6 Pt 1):2439–2448. doi: 10.1083/jcb.103.6.2439. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. McCarthy K. D., de Vellis J. Preparation of separate astroglial and oligodendroglial cell cultures from rat cerebral tissue. J Cell Biol. 1980 Jun;85(3):890–902. doi: 10.1083/jcb.85.3.890. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. McLean J. W., Vestal D. J., Cheresh D. A., Bodary S. C. cDNA sequence of the human integrin beta 5 subunit. J Biol Chem. 1990 Oct 5;265(28):17126–17131. [PubMed] [Google Scholar]
  29. Menko A. S., Boettiger D. Occupation of the extracellular matrix receptor, integrin, is a control point for myogenic differentiation. Cell. 1987 Oct 9;51(1):51–57. doi: 10.1016/0092-8674(87)90009-2. [DOI] [PubMed] [Google Scholar]
  30. Miyauchi A., Alvarez J., Greenfield E. M., Teti A., Grano M., Colucci S., Zambonin-Zallone A., Ross F. P., Teitelbaum S. L., Cheresh D. Recognition of osteopontin and related peptides by an alpha v beta 3 integrin stimulates immediate cell signals in osteoclasts. J Biol Chem. 1991 Oct 25;266(30):20369–20374. [PubMed] [Google Scholar]
  31. Moyle M., Napier M. A., McLean J. W. Cloning and expression of a divergent integrin subunit beta 8. J Biol Chem. 1991 Oct 15;266(29):19650–19658. [PubMed] [Google Scholar]
  32. Nathan C., Sanchez E. Tumor necrosis factor and CD11/CD18 (beta 2) integrins act synergistically to lower cAMP in human neutrophils. J Cell Biol. 1990 Nov;111(5 Pt 1):2171–2181. doi: 10.1083/jcb.111.5.2171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Nieke J., Schachner M. Expression of the neural cell adhesion molecules L1 and N-CAM and their common carbohydrate epitope L2/HNK-1 during development and after transection of the mouse sciatic nerve. Differentiation. 1985;30(2):141–151. doi: 10.1111/j.1432-0436.1985.tb00525.x. [DOI] [PubMed] [Google Scholar]
  34. Oz O. K., Campbell A., Tao T. W. Reduced cell adhesion to fibronectin and laminin is associated with altered glycosylation of beta 1 integrins in a weakly metastatic glycosylation mutant. Int J Cancer. 1989 Aug 15;44(2):343–347. doi: 10.1002/ijc.2910440226. [DOI] [PubMed] [Google Scholar]
  35. Palmer E. L., Rüegg C., Ferrando R., Pytela R., Sheppard D. Sequence and tissue distribution of the integrin alpha 9 subunit, a novel partner of beta 1 that is widely distributed in epithelia and muscle. J Cell Biol. 1993 Dec;123(5):1289–1297. doi: 10.1083/jcb.123.5.1289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Pardi R., Inverardi L., Rugarli C., Bender J. R. Antigen-receptor complex stimulation triggers protein kinase C-dependent CD11a/CD18-cytoskeleton association in T lymphocytes. J Cell Biol. 1992 Mar;116(5):1211–1220. doi: 10.1083/jcb.116.5.1211. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Pesheva P., Juliano R. L., Schachner M. Expression and localization of the fibronectin receptor in the mouse nervous system. J Neurosci Res. 1988 Aug;20(4):420–430. doi: 10.1002/jnr.490200404. [DOI] [PubMed] [Google Scholar]
  38. Ramaswamy H., Hemler M. E. Cloning, primary structure and properties of a novel human integrin beta subunit. EMBO J. 1990 May;9(5):1561–1568. doi: 10.1002/j.1460-2075.1990.tb08275.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Reichardt L. F., Tomaselli K. J. Extracellular matrix molecules and their receptors: functions in neural development. Annu Rev Neurosci. 1991;14:531–570. doi: 10.1146/annurev.ne.14.030191.002531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Richter J., Ng-Sikorski J., Olsson I., Andersson T. Tumor necrosis factor-induced degranulation in adherent human neutrophils is dependent on CD11b/CD18-integrin-triggered oscillations of cytosolic free Ca2+. Proc Natl Acad Sci U S A. 1990 Dec;87(23):9472–9476. doi: 10.1073/pnas.87.23.9472. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Rome L. H., Bullock P. N., Chiappelli F., Cardwell M., Adinolfi A. M., Swanson D. Synthesis of a myelin-like membrane by oligodendrocytes in culture. J Neurosci Res. 1986;15(1):49–65. doi: 10.1002/jnr.490150106. [DOI] [PubMed] [Google Scholar]
  42. Ruoslahti E., Pierschbacher M. D. Arg-Gly-Asp: a versatile cell recognition signal. Cell. 1986 Feb 28;44(4):517–518. doi: 10.1016/0092-8674(86)90259-x. [DOI] [PubMed] [Google Scholar]
  43. Ruoslahti E., Pierschbacher M. D. New perspectives in cell adhesion: RGD and integrins. Science. 1987 Oct 23;238(4826):491–497. doi: 10.1126/science.2821619. [DOI] [PubMed] [Google Scholar]
  44. Sheppard D., Rozzo C., Starr L., Quaranta V., Erle D. J., Pytela R. Complete amino acid sequence of a novel integrin beta subunit (beta 6) identified in epithelial cells using the polymerase chain reaction. J Biol Chem. 1990 Jul 15;265(20):11502–11507. [PubMed] [Google Scholar]
  45. Song W. K., Wang W., Foster R. F., Bielser D. A., Kaufman S. J. H36-alpha 7 is a novel integrin alpha chain that is developmentally regulated during skeletal myogenesis. J Cell Biol. 1992 May;117(3):643–657. doi: 10.1083/jcb.117.3.643. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Suzuki S., Naitoh Y. Amino acid sequence of a novel integrin beta 4 subunit and primary expression of the mRNA in epithelial cells. EMBO J. 1990 Mar;9(3):757–763. doi: 10.1002/j.1460-2075.1990.tb08170.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Tawil N. J., Houde M., Blacher R., Esch F., Reichardt L. F., Turner D. C., Carbonetto S. Alpha 1 beta 1 integrin heterodimer functions as a dual laminin/collagen receptor in neural cells. Biochemistry. 1990 Jul 10;29(27):6540–6544. doi: 10.1021/bi00479a028. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Vogel B. E., Tarone G., Giancotti F. G., Gailit J., Ruoslahti E. A novel fibronectin receptor with an unexpected subunit composition (alpha v beta 1). J Biol Chem. 1990 Apr 15;265(11):5934–5937. [PubMed] [Google Scholar]
  49. Wadsworth S., Halvorson M. J., Chang A. C., Coligan J. E. Multiple changes in VLA protein glycosylation, expression, and function occur during mouse T cell ontogeny. J Immunol. 1993 Feb 1;150(3):847–857. [PubMed] [Google Scholar]
  50. Werb Z., Tremble P. M., Behrendtsen O., Crowley E., Damsky C. H. Signal transduction through the fibronectin receptor induces collagenase and stromelysin gene expression. J Cell Biol. 1989 Aug;109(2):877–889. doi: 10.1083/jcb.109.2.877. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Yuan Q., Jiang W. M., Leung E., Hollander D., Watson J. D., Krissansen G. W. Molecular cloning of the mouse integrin beta 7 subunit. J Biol Chem. 1992 Apr 15;267(11):7352–7358. [PubMed] [Google Scholar]

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

RESOURCES