Abstract
A pepsin-resistant triple helical domain (chain 50,000 Mr) of type VIII collagen was isolated from bovine corneal Descemet's membrane and used as an immunogen for the production of mAbs. An antibody was selected for biochemical and tissue immunofluorescence studies which reacted both with Descemet's membrane and with type VIII collagen 50,000-Mr polypeptides by competition ELISA and immunoblotting. This antibody exhibited no crossreactivity with collagen types I-VI by competition ELISA. The mAb specifically precipitated a high molecular mass component of type VIII collagen (EC2, of chain 125,000 Mr) from the culture medium of subconfluent bovine corneal endothelial cells metabolically labeled for 24 h. In contrast, confluent cells in the presence of FCS and isotope for 7 d secreted a collagenous component of chain 60,000 Mr that did not react with the anti-type VIII collagen IgG. Type VIII collagen therefore appears to be synthesized as a discontinuous triple helical molecule with a predominant chain 125,000 Mr by subconfluent, proliferating cells in culture. Immunofluorescence studies with the mAb showed that type VIII collagen was deposited as fibrils in the extracellular matrix of corneal endothelial cells. In the fetal calf, type VIII collagen was absent from basement membranes and was found in a limited number of tissues. In addition to the linear staining pattern observed in the Descemet's membrane, type VIII collagen was found in highly fibrillar arrays in the ocular sclera, in the meninges surrounding brain, spinal cord, and optic nerve, and in periosteum and perichondrium. Fine fibrils were evident in the white matter of spinal cord, whereas a more generalized staining was apparent in the matrices of cartilage and bone. Despite attempts to unmask the epitope, type VIII collagen was not found in aorta, kidney, lung, liver, skin, and ligament. We conclude that this unusual collagen is a component of certain specialized extracellular matrices, several of which are derived from the neural crest.
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- Alitalo K., Bornstein P., Vaheri A., Sage H. Biosynthesis of an unusual collagen type by human astrocytoma cells in vitro. J Biol Chem. 1983 Feb 25;258(4):2653–2661. [PubMed] [Google Scholar]
- Bahn C. F., Falls H. F., Varley G. A., Meyer R. F., Edelhauser H. F., Bourne W. M. Classification of corneal endothelial disorders based on neural crest origin. Ophthalmology. 1984 Jun;91(6):558–563. doi: 10.1016/s0161-6420(84)34249-x. [DOI] [PubMed] [Google Scholar]
- Benya P. D., Padilla S. R. Isolation and characterization of type VIII collagen synthesized by cultured rabbit corneal endothelial cells. A conventional structure replaces the interrupted-helix model. J Biol Chem. 1986 Mar 25;261(9):4160–4169. [PubMed] [Google Scholar]
- Carlson E. C., Brendel K., Hjelle J. T., Meezan E. Ultrastructural and biochemical analyses of isolated basement membranes from kidney glomeruli and tubules and brain and retinal microvessels. J Ultrastruct Res. 1978 Jan;62(1):26–53. doi: 10.1016/s0022-5320(78)80028-8. [DOI] [PubMed] [Google Scholar]
- Engvall E., Perlmann P. Enzyme-linked immunosorbent assay, Elisa. 3. Quantitation of specific antibodies by enzyme-labeled anti-immunoglobulin in antigen-coated tubes. J Immunol. 1972 Jul;109(1):129–135. [PubMed] [Google Scholar]
- Fitch J. M., Linsenmayer T. F. Monoclonal antibody analysis of ocular basement membranes during development. Dev Biol. 1983 Jan;95(1):137–153. doi: 10.1016/0012-1606(83)90013-1. [DOI] [PubMed] [Google Scholar]
- Hall B. K. Matrices control the differentiation of cartilage and bone. Prog Clin Biol Res. 1984;151:147–169. [PubMed] [Google Scholar]
- Hay E. D. Development of the vertebrate cornea. Int Rev Cytol. 1980;63:263–322. doi: 10.1016/s0074-7696(08)61760-x. [DOI] [PubMed] [Google Scholar]
- JAKUS M. A. Studies on the cornea. II. The fine structure of Descement's membrane. J Biophys Biochem Cytol. 1956 Jul 25;2(4 Suppl):243–252. doi: 10.1083/jcb.2.4.243. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kapoor R., Bornstein P., Sage E. H. Type VIII collagen from bovine Descemet's membrane: structural characterization of a triple-helical domain. Biochemistry. 1986 Jul 1;25(13):3930–3937. doi: 10.1021/bi00361a029. [DOI] [PubMed] [Google Scholar]
- Kefalides N. A., Cameron J. D., Tomichek E. A., Yanoff M. Biosynthesis of basement membrane collagen by rabbit corneal endothelium in vitro. J Biol Chem. 1976 Feb 10;251(3):730–733. [PubMed] [Google Scholar]
- Kenney M. C., Labermeier U., Hinds D., Waring G. O., 3rd Characterization of the Descemet's membrane/posterior collagenous layer isolated from Fuchs' endothelial dystrophy corneas. Exp Eye Res. 1984 Sep;39(3):267–277. doi: 10.1016/0014-4835(84)90015-0. [DOI] [PubMed] [Google Scholar]
- Köhler G., Howe S. C., Milstein C. Fusion between immunoglobulin-secreting and nonsecreting myeloma cell lines. Eur J Immunol. 1976 Apr;6(4):292–295. doi: 10.1002/eji.1830060411. [DOI] [PubMed] [Google Scholar]
- Labermeier U., Kenney M. C. The presence of EC collagen and type IV collagen in bovine Descemet's membranes. Biochem Biophys Res Commun. 1983 Oct 31;116(2):619–625. doi: 10.1016/0006-291x(83)90569-7. [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]
- Linsenmayer T. F., Fitch J. M., Schmid T. M., Zak N. B., Gibney E., Sanderson R. D., Mayne R. Monoclonal antibodies against chicken type V collagen: production, specificity, and use for immunocytochemical localization in embryonic cornea and other organs. J Cell Biol. 1983 Jan;96(1):124–132. doi: 10.1083/jcb.96.1.124. [DOI] [PMC free article] [PubMed] [Google Scholar]
- MacCallum D. K., Lillie J. H., Scaletta L. J., Occhino J. C., Frederick W. G., Ledbetter S. R. Bovine corneal endothelium in vitro. Elaboration and organization and of a basement membrane. Exp Cell Res. 1982 May;139(1):1–13. doi: 10.1016/0014-4827(82)90313-5. [DOI] [PubMed] [Google Scholar]
- Maumenee I. H. The cornea in connective tissue diseases. Ophthalmology. 1978 Oct;85(10):1014–1017. doi: 10.1016/s0161-6420(78)35591-3. [DOI] [PubMed] [Google Scholar]
- McGill H. C., Jr, McMahan C. A., Kruski A. W., Mott G. E. Relationship of lipoprotein cholesterol concentrations to experimental atherosclerosis in baboons. Arteriosclerosis. 1981 Jan-Feb;1(1):3–12. doi: 10.1161/01.atv.1.1.3. [DOI] [PubMed] [Google Scholar]
- Sage H., Balian G., Vogel A. M., Bornstein P. Type VIII collagen. Synthesis by normal and malignant cells in culture. Lab Invest. 1984 Feb;50(2):219–231. [PubMed] [Google Scholar]
- Sage H., Pritzl P., Bornstein P. A unique, pepsin-sensitive collagen synthesized by aortic endothelial cells in culture. Biochemistry. 1980 Dec 9;19(25):5747–5755. doi: 10.1021/bi00566a013. [DOI] [PubMed] [Google Scholar]
- Sage H., Trüeb B., Bornstein P. Biosynthetic and structural properties of endothelial cell type VIII collagen. J Biol Chem. 1983 Nov 10;258(21):13391–13401. [PubMed] [Google Scholar]
- Sakai L. Y., Engvall E., Hollister D. W., Burgeson R. E. Production and characterization of a monoclonal antibody to human Type IV collagen. Am J Pathol. 1982 Sep;108(3):310–318. [PMC free article] [PubMed] [Google Scholar]
- Sakai L. Y., Keene D. R., Engvall E. Fibrillin, a new 350-kD glycoprotein, is a component of extracellular microfibrils. J Cell Biol. 1986 Dec;103(6 Pt 1):2499–2509. doi: 10.1083/jcb.103.6.2499. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sawada H., Konomi H., Nagai Y. The basement membrane of bovine corneal endothelial cells in culture with beta-aminopropionitrile: biosynthesis of hexagonal lattices composed of a 160 nm dumbbell-shaped structure. Eur J Cell Biol. 1984 Nov;35(2):226–234. [PubMed] [Google Scholar]
- Small R. K., Riddle P., Noble M. Evidence for migration of oligodendrocyte--type-2 astrocyte progenitor cells into the developing rat optic nerve. Nature. 1987 Jul 9;328(6126):155–157. doi: 10.1038/328155a0. [DOI] [PubMed] [Google Scholar]
- Thorogood P., Bee J., von der Mark K. Transient expression of collagen type II at epitheliomesenchymal interfaces during morphogenesis of the cartilaginous neurocranium. Dev Biol. 1986 Aug;116(2):497–509. doi: 10.1016/0012-1606(86)90150-8. [DOI] [PubMed] [Google Scholar]
- Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
- von der Mark K., von der Mark H., Timpl R., Trelstad R. L. Immunofluorescent localization of collagen types I, II, and III in the embryonic chick eye. Dev Biol. 1977 Aug;59(1):75–85. doi: 10.1016/0012-1606(77)90241-x. [DOI] [PubMed] [Google Scholar]