Abstract
To determine whether specific asparagine-linked (N-linked) oligosaccharides present in cell surface glycoproteins are required for cell-cell interactions within the peripheral nervous system, we have used castanospermine to inhibit maturation of N-linked sugars in cell cultures of neurons or neurons plus Schwann cells. Maximally 10-15% of the N-linked oligosaccharides on neuronal proteins have normal structure when cells are cultured in the presence of 250 micrograms/ml castanospermine; the remaining oligosaccharides are present as immature carbohydrate chains not normally found in these glycoproteins. Although cultures were treated for 2 wk with castanospermine, cells always remained viable and appeared healthy. We have analyzed several biological responses of embryonic dorsal root ganglion neurons, with or without added purified populations of Schwann cells, in the presence of castanospermine. We have observed that a normal complement of mature, N- linked sugars are not required for neurite outgrowth, neuron-Schwann cell adhesion, neuron-induced Schwann cell proliferation, or ensheathment of neurites by Schwann cells. Treatment of neuronal cultures with castanospermine increases the propensity of neurites to fasciculate. Extracellular matrix deposition by Schwann cells and myelination of neurons by Schwann cells are greatly diminished in the presence of castanospermine as assayed by electron microscopy and immunocytochemistry, suggesting that specific N-linked oligosaccharides are required for the expression of these cellular functions.
Full Text
The Full Text of this article is available as a PDF (1.9 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Aguayo A. J., Charron L., Bray G. M. Potential of Schwann cells from unmyelinated nerves to produce myelin: a quantitative ultrastructural and radiographic study. J Neurocytol. 1976 Oct;5(8):565–573. doi: 10.1007/BF01175570. [DOI] [PubMed] [Google Scholar]
- Argiro V., Johnson M. I. Patterns and kinetics of neurite extension from sympathetic neurons in culture are age dependent. J Neurosci. 1982 Apr;2(4):503–512. doi: 10.1523/JNEUROSCI.02-04-00503.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bottenstein J. E., Sato G. H. Growth of a rat neuroblastoma cell line in serum-free supplemented medium. Proc Natl Acad Sci U S A. 1979 Jan;76(1):514–517. doi: 10.1073/pnas.76.1.514. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bunge M. B., Williams A. K., Wood P. M. Neuron-Schwann cell interaction in basal lamina formation. Dev Biol. 1982 Aug;92(2):449–460. doi: 10.1016/0012-1606(82)90190-7. [DOI] [PubMed] [Google Scholar]
- Bunge R. P., Bunge M. B., Eldridge C. F. Linkage between axonal ensheathment and basal lamina production by Schwann cells. Annu Rev Neurosci. 1986;9:305–328. doi: 10.1146/annurev.ne.09.030186.001513. [DOI] [PubMed] [Google Scholar]
- Bunge R. P., Wood P. Studies on the transplantation of spinal cord tissue in the rat. I. The development of a culture system for hemisections of embryonic spinal cord. Brain Res. 1973 Jul 27;57(2):261–276. doi: 10.1016/0006-8993(73)90135-2. [DOI] [PubMed] [Google Scholar]
- Carey D. J., Bunge R. P. Factors influencing the release of proteins by cultured Schwann cells. J Cell Biol. 1981 Dec;91(3 Pt 1):666–672. doi: 10.1083/jcb.91.3.666. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Carey D. J., Eldridge C. F., Cornbrooks C. J., Timpl R., Bunge R. P. Biosynthesis of type IV collagen by cultured rat Schwann cells. J Cell Biol. 1983 Aug;97(2):473–479. doi: 10.1083/jcb.97.2.473. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Carlin B., Jaffe R., Bender B., Chung A. E. Entactin, a novel basal lamina-associated sulfated glycoprotein. J Biol Chem. 1981 May 25;256(10):5209–5214. [PubMed] [Google Scholar]
- Chuong C. M., Edelman G. M. Alterations in neural cell adhesion molecules during development of different regions of the nervous system. J Neurosci. 1984 Sep;4(9):2354–2368. doi: 10.1523/JNEUROSCI.04-09-02354.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Clark C. C. The distribution and initial characterization of oligosaccharide units on the COOH-terminal propeptide extensions of the pro-alpha 1 and pro-alpha 2 chains of type I procollagen. J Biol Chem. 1979 Nov 10;254(21):10798–10802. [PubMed] [Google Scholar]
- Cornbrooks C. J., Carey D. J., McDonald J. A., Timpl R., Bunge R. P. In vivo and in vitro observations on laminin production by Schwann cells. Proc Natl Acad Sci U S A. 1983 Jun;80(12):3850–3854. doi: 10.1073/pnas.80.12.3850. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cunningham B. A., Hoffman S., Rutishauser U., Hemperly J. J., Edelman G. M. Molecular topography of the neural cell adhesion molecule N-CAM: surface orientation and location of sialic acid-rich and binding regions. Proc Natl Acad Sci U S A. 1983 May;80(10):3116–3120. doi: 10.1073/pnas.80.10.3116. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Elbein A. D. Inhibitors of the biosynthesis and processing of N-linked oligosaccharides. CRC Crit Rev Biochem. 1984;16(1):21–49. doi: 10.3109/10409238409102805. [DOI] [PubMed] [Google Scholar]
- Eldridge C. F., Sanes J. R., Chiu A. Y., Bunge R. P., Cornbrooks C. J. Basal lamina-associated heparan sulphate proteoglycan in the rat PNS: characterization and localization using monoclonal antibodies. J Neurocytol. 1986 Feb;15(1):37–51. doi: 10.1007/BF02057903. [DOI] [PubMed] [Google Scholar]
- Florman H. M., Wassarman P. M. O-linked oligosaccharides of mouse egg ZP3 account for its sperm receptor activity. Cell. 1985 May;41(1):313–324. doi: 10.1016/0092-8674(85)90084-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lodish H. F., Kong N. Glucose removal from N-linked oligosaccharides is required for efficient maturation of certain secretory glycoproteins from the rough endoplasmic reticulum to the Golgi complex. J Cell Biol. 1984 May;98(5):1720–1729. doi: 10.1083/jcb.98.5.1720. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lopez L. C., Bayna E. M., Litoff D., Shaper N. L., Shaper J. H., Shur B. D. Receptor function of mouse sperm surface galactosyltransferase during fertilization. J Cell Biol. 1985 Oct;101(4):1501–1510. doi: 10.1083/jcb.101.4.1501. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Moya F., Bunge M. B., Bunge R. P. Schwann cells proliferate but fail to differentiate in defined medium. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6902–6906. doi: 10.1073/pnas.77.11.6902. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pfenninger K. H., Maylié-Pfenninger M. F. Lectin labeling of sprouting neurons. II. Relative movement and appearance of glycoconjugates during plasmalemmal expansion. J Cell Biol. 1981 Jun;89(3):547–559. doi: 10.1083/jcb.89.3.547. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ranscht B., Clapshaw P. A., Price J., Noble M., Seifert W. Development of oligodendrocytes and Schwann cells studied with a monoclonal antibody against galactocerebroside. Proc Natl Acad Sci U S A. 1982 Apr;79(8):2709–2713. doi: 10.1073/pnas.79.8.2709. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ratner N., Bunge R. P., Glaser L. A neuronal cell surface heparan sulfate proteoglycan is required for dorsal root ganglion neuron stimulation of Schwann cell proliferation. J Cell Biol. 1985 Sep;101(3):744–754. doi: 10.1083/jcb.101.3.744. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ratner N., Glaser L., Bunge R. P. PC12 cells as a source of neurite-derived cell surface mitogen, which stimulates Schwann cell division. J Cell Biol. 1984 Mar;98(3):1150–1155. doi: 10.1083/jcb.98.3.1150. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Repp R., Tamura T., Boschek C. B., Wege H., Schwarz R. T., Niemann H. The effects of processing inhibitors of N-linked oligosaccharides on the intracellular migration of glycoprotein E2 of mouse hepatitis virus and the maturation of coronavirus particles. J Biol Chem. 1985 Dec 15;260(29):15873–15879. doi: 10.1016/S0021-9258(17)36339-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Roseman S. The synthesis of complex carbohydrates by multiglycosyltransferase systems and their potential function in intercellular adhesion. Chem Phys Lipids. 1970 Oct;5(1):270–297. doi: 10.1016/0009-3084(70)90024-1. [DOI] [PubMed] [Google Scholar]
- Roufa D. G., Johnson M. I., Bunge M. B. Influence of ganglion age, nonneuronal cells and substratum on neurite outgrowth in culture. Dev Biol. 1983 Sep;99(1):225–239. doi: 10.1016/0012-1606(83)90271-3. [DOI] [PubMed] [Google Scholar]
- Rutishauser U., Gall W. E., Edelman G. M. Adhesion among neural cells of the chick embryo. IV. Role of the cell surface molecule CAM in the formation of neurite bundles in cultures of spinal ganglia. J Cell Biol. 1978 Nov;79(2 Pt 1):382–393. doi: 10.1083/jcb.79.2.382. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rutishauser U., Watanabe M., Silver J., Troy F. A., Vimr E. R. Specific alteration of NCAM-mediated cell adhesion by an endoneuraminidase. J Cell Biol. 1985 Nov;101(5 Pt 1):1842–1849. doi: 10.1083/jcb.101.5.1842. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Salzer J. L., Bunge R. P., Glaser L. Studies of Schwann cell proliferation. III. Evidence for the surface localization of the neurite mitogen. J Cell Biol. 1980 Mar;84(3):767–778. doi: 10.1083/jcb.84.3.767. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Salzer J. L., Bunge R. P. Studies of Schwann cell proliferation. I. An analysis in tissue culture of proliferation during development, Wallerian degeneration, and direct injury. J Cell Biol. 1980 Mar;84(3):739–752. doi: 10.1083/jcb.84.3.739. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Salzer J. L., Williams A. K., Glaser L., Bunge R. P. Studies of Schwann cell proliferation. II. Characterization of the stimulation and specificity of the response to a neurite membrane fraction. J Cell Biol. 1980 Mar;84(3):753–766. doi: 10.1083/jcb.84.3.753. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schlesinger S., Malfer C., Schlesinger M. J. The formation of vesicular stomatitis virus (San Juan strain) becomes temperature-sensitive when glucose residues are retained on the oligosaccharides of the glycoprotein. J Biol Chem. 1984 Jun 25;259(12):7597–7601. [PubMed] [Google Scholar]
- Sobue G., Pleasure D. Adhesion of axolemmal fragments to Schwann cells: a signal- and target-specific process closely linked to axolemmal induction of Schwann cell mitosis. J Neurosci. 1985 Feb;5(2):379–387. doi: 10.1523/JNEUROSCI.05-02-00379.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sobue G., Pleasure D. Schwann cell galactocerebroside induced by derivatives of adenosine 3',5'-monophosphate. Science. 1984 Apr 6;224(4644):72–74. doi: 10.1126/science.6322307. [DOI] [PubMed] [Google Scholar]
- Stallcup W. B., Beasley L. L., Levine J. M. Antibody against nerve growth factor-inducible large external (NILE) glycoprotein labels nerve fiber tracts in the developing rat nervous system. J Neurosci. 1985 Apr;5(4):1090–1101. doi: 10.1523/JNEUROSCI.05-04-01090.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stallcup W. B., Beasley L. Involvement of the nerve growth factor-inducible large external glycoprotein (NILE) in neurite fasciculation in primary cultures of rat brain. Proc Natl Acad Sci U S A. 1985 Feb;82(4):1276–1280. doi: 10.1073/pnas.82.4.1276. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stoolman L. M., Tenforde T. S., Rosen S. D. Phosphomannosyl receptors may participate in the adhesive interaction between lymphocytes and high endothelial venules. J Cell Biol. 1984 Oct;99(4 Pt 1):1535–1540. doi: 10.1083/jcb.99.4.1535. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Timpl R., Rohde H., Robey P. G., Rennard S. I., Foidart J. M., Martin G. R. Laminin--a glycoprotein from basement membranes. J Biol Chem. 1979 Oct 10;254(19):9933–9937. [PubMed] [Google Scholar]
- Timpl R., Wiedemann H., van Delden V., Furthmayr H., Kühn K. A network model for the organization of type IV collagen molecules in basement membranes. Eur J Biochem. 1981 Nov;120(2):203–211. doi: 10.1111/j.1432-1033.1981.tb05690.x. [DOI] [PubMed] [Google Scholar]
- Trapp B. D., Quarles R. H., Suzuki K. Immunocytochemical studies of quaking mice support a role for the myelin-associated glycoprotein in forming and maintaining the periaxonal space and periaxonal cytoplasmic collar of myelinating Schwann cells. J Cell Biol. 1984 Aug;99(2):594–606. doi: 10.1083/jcb.99.2.594. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Webster H. D., Martin R., O'Connell M. F. The relationships between interphase Schwann cells and axons before myelination: a quantitative electron microscopic study. Dev Biol. 1973 Jun;32(2):401–416. doi: 10.1016/0012-1606(73)90250-9. [DOI] [PubMed] [Google Scholar]
- Weinberg H. J., Spencer P. S. Studies on the control of myelinogenesis. I. Myelination of regenerating axons after entry into a foreign unmyelinated nerve. J Neurocytol. 1975 Aug;4(4):395–418. doi: 10.1007/BF01261372. [DOI] [PubMed] [Google Scholar]
- Wice B. M., Kennell D. E. Sugar-free growth of mammalian cells on some ribonucleosides but not on others. J Biol Chem. 1983 Nov 10;258(21):13134–13140. [PubMed] [Google Scholar]
- Wood P. M. Separation of functional Schwann cells and neurons from normal peripheral nerve tissue. Brain Res. 1976 Oct 22;115(3):361–375. doi: 10.1016/0006-8993(76)90355-3. [DOI] [PubMed] [Google Scholar]
- Yavin E., Richter-Landsberg C., Duksin D., Yavin Z. Tunicamycin blocks neuritogenesis and glucosamine labeling of gangliosides in developing cerebral neuron cultures. Proc Natl Acad Sci U S A. 1984 Sep;81(18):5638–5642. doi: 10.1073/pnas.81.18.5638. [DOI] [PMC free article] [PubMed] [Google Scholar]