Abstract
We have obtained evidence that rat Schwann cells synthesize and secrete type IV procollagen. Metabolic labeling of primary cultures of Schwann cells plus neurons and analysis by SDS PAGE revealed the presence of a closely spaced pair of polypeptides in the medium of these cultures that (a) were susceptible to digestion by purified bacterial collagenase, (b) co-migrated with type IV procollagen secreted by rat parietal endoderm cells, and (c) were specifically immunoprecipitated by antibodies against mouse type IV collagen. Limited pepsin digestion of metabolically labeled medium or cell layers produced a pepsin- resistant fragment characteristic of pro-alpha 1(IV) chains. Removal of neuronal cell bodies from the cultures immediately before labeling did not reduce the amount of type IV procollagen detected in the medium. This indicated that Schwann cells, not neurons, were responsible for synthesis of type IV procollagen. We believe type IV procollagen is a major constituent of the Schwann-cell extracellular matrix based upon (a) its presence in a detergent-insoluble matrix preparation, (b) its presence in the cell layer of the cultures in a state in which it can be removed by brief treatment with bacterial collagenase or trypsin, and (c) positive immunofluorescence of Schwann cell-neuron cultures with anti-type-IV collagen antibodies. Secretion of type IV procollagen was substantially reduced when Schwann cells were maintained in the absence of neurons. This observation may account for the previously reported finding that Schwann cells assemble a basal lamina only when co-cultured with neurons (Bunge, M. B., A. K. Williams, and P. M. Wood, 1982, Dev. Biol., 92:449).
Full Text
The Full Text of this article is available as a PDF (1.8 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Aguayo A. J., Epps J., Charron L., Bray G. M. Multipotentiality of Schwann cells in cross-anastomosed and grafted myelinated and unmyelinated nerves: quantitative microscopy and radioautography. Brain Res. 1976 Mar 5;104(1):1–20. doi: 10.1016/0006-8993(76)90643-0. [DOI] [PubMed] [Google Scholar]
- Alitalo K., Kurkinen M., Vaheri A., Virtanen I., Rohde H., Timpl R. Basal lamina glycoproteins are produced by neuroblastoma cells. Nature. 1980 Oct 2;287(5781):465–466. doi: 10.1038/287465a0. [DOI] [PubMed] [Google Scholar]
- Alitalo K., Vaheri A., Krieg T., Timpl R. Biosynthesis of two subunits of type IV procollagen and of other basement membrane proteins by a human tumor cell line. Eur J Biochem. 1980 Aug;109(1):247–255. doi: 10.1111/j.1432-1033.1980.tb04790.x. [DOI] [PubMed] [Google Scholar]
- Billings-Gagliardi S., Webster H. F., O'Connell M. F. In vivo and electron microscopic observations on Schwann cells in developing tadpole nerve fibers. Am J Anat. 1974 Nov;141(3):375–391. doi: 10.1002/aja.1001410308. [DOI] [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 M. B., Williams A. K., Wood P. M., Uitto J., Jeffrey J. J. Comparison of nerve cell and nerve cell plus Schwann cell cultures, with particular emphasis on basal lamina and collagen formation. J Cell Biol. 1980 Jan;84(1):184–202. doi: 10.1083/jcb.84.1.184. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bächinger H. P., Fessler L. I., Fessler J. H. Mouse procollagen IV. Characterization and supramolecular association. J Biol Chem. 1982 Aug 25;257(16):9796–9803. [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]
- Clark C. C., Tomichek E. A., Koszalka T. R., Minor R. R., Kefalides N. A. The embryonic rat parietal yolk sac. The role of the parietal endoderm in the biosynthesis of basement membrane collagen and glycoprotein in vitro. J Biol Chem. 1975 Jul 10;250(13):5259–5267. [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]
- Courtoy P. J., Timpl R., Farquhar M. G. Comparative distribution of laminin, type IV collagen, and fibronectin in the rat glomerulus. J Histochem Cytochem. 1982 Sep;30(9):874–886. doi: 10.1177/30.9.7130672. [DOI] [PubMed] [Google Scholar]
- Crouch E., Sage H., Bornstein P. Structural basis for apparent heterogeneity of collagens in human basement membranes: type IV procollagen contains two distinct chains. Proc Natl Acad Sci U S A. 1980 Feb;77(2):745–749. doi: 10.1073/pnas.77.2.745. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Heathcote J. G., Grant M. E. The molecular organization of basement membranes. Int Rev Connect Tissue Res. 1981;9:191–264. doi: 10.1016/b978-0-12-363709-3.50011-5. [DOI] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Laurie G. W., Leblond C. P., Martin G. R. Localization of type IV collagen, laminin, heparan sulfate proteoglycan, and fibronectin to the basal lamina of basement membranes. J Cell Biol. 1982 Oct;95(1):340–344. doi: 10.1083/jcb.95.1.340. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Liotta L. A., Wicha M. S., Foidart J. M., Rennard S. I., Garbisa S., Kidwell W. R. Hormonal requirements for basement membrane collagen deposition by cultured rat mammary epithelium. Lab Invest. 1979 Dec;41(6):511–518. [PubMed] [Google Scholar]
- Meezan E., Hjelle J. T., Brendel K., Carlson E. C. A simple, versatile, nondisruptive method for the isolation of morphologically and chemically pure basement membranes from several tissues. Life Sci. 1975 Dec 1;17(11):1721–1732. doi: 10.1016/0024-3205(75)90119-8. [DOI] [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]
- Oberbäumer I., Wiedemann H., Timpl R., Kühn K. Shape and assembly of type IV procollagen obtained from cell culture. EMBO J. 1982;1(7):805–810. doi: 10.1002/j.1460-2075.1982.tb01251.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Okada E., Bunge R. P., Bunge M. B. Abnormalities expressed in long-term cultures of dorsal root ganglia from the dystrophic mouse. Brain Res. 1980 Aug 4;194(2):455–470. doi: 10.1016/0006-8993(80)91225-1. [DOI] [PubMed] [Google Scholar]
- Prehm P., Dessau W., Timpl R. Rates of synthesis of basement membrane proteins by differentiating teratocarcinoma stem cells and their modulation by hormones. Connect Tissue Res. 1982;10(3-4):275–285. doi: 10.3109/03008208209008053. [DOI] [PubMed] [Google Scholar]
- Roll F. J., Madri J. A., Albert J., Furthmayr H. Codistribution of collagen types IV and AB2 in basement membranes and mesangium of the kidney. an immunoferritin study of ultrathin frozen sections. J Cell Biol. 1980 Jun;85(3):597–616. doi: 10.1083/jcb.85.3.597. [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]
- Sanes J. R. Laminin, fibronectin, and collagen in synaptic and extrasynaptic portions of muscle fiber basement membrane. J Cell Biol. 1982 May;93(2):442–451. doi: 10.1083/jcb.93.2.442. [DOI] [PMC free article] [PubMed] [Google Scholar]
- THOMAS P. K. CHANGES IN THE ENDONEURIAL SHEATHS OF PERIPHERAL MYELINATED NERVE FIBRES DURING WALLERIAN DEGENERATION. J Anat. 1964 Apr;98:175–182. [PMC free article] [PubMed] [Google Scholar]
- Timpl R., Martin G. R., Bruckner P., Wick G., Wiedemann H. Nature of the collagenous protein in a tumor basement membrane. Eur J Biochem. 1978 Mar;84(1):43–52. doi: 10.1111/j.1432-1033.1978.tb12139.x. [DOI] [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]
- Tryggvason K., Robey P. G., Martin G. R. Biosynthesis of type IV procollagens. Biochemistry. 1980 Apr 1;19(7):1284–1289. doi: 10.1021/bi00548a003. [DOI] [PubMed] [Google Scholar]
- Wood P. M., Bunge R. P. Evidence that sensory axons are mitogenic for Schwann cells. Nature. 1975 Aug 21;256(5519):662–664. doi: 10.1038/256662a0. [DOI] [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]