Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1986 Dec 1;103(6):2475–2487. doi: 10.1083/jcb.103.6.2475

Conditioning of native substrates by chondroitin sulfate proteoglycans during cardiac mesenchymal cell migration

PMCID: PMC2114578  PMID: 3782305

Abstract

It is generally proposed that embryonic mesenchymal cells use sulfated macromolecules during in situ migration. Attempts to resolve the molecular mechanisms for this hypothesis using planar substrates have been met with limited success. In the present study, we provide evidence that the functional significance of certain sulfated macromolecules during mesenchyme migration required the presence of the endogenous migratory template; i.e., native collagen fibrils. Using three-dimensional collagen gel lattices and whole embryo culture procedures to produce metabolically labeled sulfated macromolecules in embryonic chick cardiac tissue, we show that these molecules were primarily proteoglycan (PG) in nature and that their distribution was class specific; i.e., heparan sulfate PG, the minor labeled component (15%), remained pericellular while chondroitin sulfate (CS) PG, the predominately labeled PG (85%), was associated with collagen fibrils as "trails" of 50-60-nm particles when viewed by scanning electron microscopy. Progressive "conditioning" of collagen with CS-PG inhibited the capacity of the template to support subsequent cell migration. Lastly, metabolically labeled, PG-derived CS chains were compared with respect to degree of sulfation in either the C-6 or C-4 position by chromatographic separation of chondroitinase AC digestion products. Results from temporal and regional comparisons of in situ-labeled PGs indicated a positive correlation between the presence of mesenchyme and an enrichment of disaccharide-4S relative to that from regions lacking mesenchyme (i.e., principally myocardial tissue). The suggestion of a mesenchyme-specific CS-PG was substantiated by similarly examining the PGs synthesized solely by cardiac mesenchymal cells migrating within hydrated collagen lattice in culture. These data were incorporated into a model of "substratum conditioning" which provides a molecular mechanism by which secretion of mesenchyme-specific CS-PGs not only provides for directed and sustained cell movement, but ultimately inhibits migration of the cell population as a whole.

Full Text

The Full Text of this article is available as a PDF (5.1 MB).

Selected References

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

  1. Abatangelo G., Cortivo R., Martelli M., Vecchia P. Cell detachment mediated by hyaluronic acid. Exp Cell Res. 1982 Jan;137(1):73–78. doi: 10.1016/0014-4827(82)90009-x. [DOI] [PubMed] [Google Scholar]
  2. Avnur Z., Geiger B. Immunocytochemical localization of native chondroitin-sulfate in tissues and cultured cells using specific monoclonal antibody. Cell. 1984 Oct;38(3):811–822. doi: 10.1016/0092-8674(84)90276-9. [DOI] [PubMed] [Google Scholar]
  3. Bernanke D. H., Markwald R. R. Migratory behavior of cardiac cushion tissue cells in a collagen-lattice culture system. Dev Biol. 1982 Jun;91(2):235–245. doi: 10.1016/0012-1606(82)90030-6. [DOI] [PubMed] [Google Scholar]
  4. Carlson D. M. Structures and immunochemical properties of oligosaccharides isolated from pig submaxillary mucins. J Biol Chem. 1968 Feb 10;243(3):616–626. [PubMed] [Google Scholar]
  5. Carrino D. A., Caplan A. I. Isolation and partial characterization of high-buoyant-density proteoglycans synthesized in ovo by embryonic chick skeletal muscle and heart. J Biol Chem. 1984 Oct 25;259(20):12419–12430. [PubMed] [Google Scholar]
  6. Chen W. T., Hasegawa E., Hasegawa T., Weinstock C., Yamada K. M. Development of cell surface linkage complexes in cultured fibroblasts. J Cell Biol. 1985 Apr;100(4):1103–1114. doi: 10.1083/jcb.100.4.1103. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Damsky C. H., Knudsen K. A., Bradley D., Buck C. A., Horwitz A. F. Distribution of the cell substratum attachment (CSAT) antigen on myogenic and fibroblastic cells in culture. J Cell Biol. 1985 May;100(5):1528–1539. doi: 10.1083/jcb.100.5.1528. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fisher M., Solursh M. The influence of the substratum on mesenchyme spreading in vitro. Exp Cell Res. 1979 Oct 1;123(1):1–13. doi: 10.1016/0014-4827(79)90416-6. [DOI] [PubMed] [Google Scholar]
  9. Fitzharris T. P., Markwald R. R. Cellular migration through the cardiac jelly matrix: a stereoanalysis by high-voltage electron microscopy. Dev Biol. 1982 Aug;92(2):315–329. doi: 10.1016/0012-1606(82)90178-6. [DOI] [PubMed] [Google Scholar]
  10. Giancotti F. G., Tarone G., Knudsen K., Damsky C., Comoglio P. M. Cleavage of a 135 kD cell surface glycoprotein correlates with loss of fibroblast adhesion to fibronectin. Exp Cell Res. 1985 Jan;156(1):182–190. doi: 10.1016/0014-4827(85)90272-1. [DOI] [PubMed] [Google Scholar]
  11. Icardo J. M., Manasek F. J. An indirect immunofluorescence study of the distribution of fibronectin during the formation of the cushion tissue mesenchyme in the embryonic heart. Dev Biol. 1984 Feb;101(2):336–345. doi: 10.1016/0012-1606(84)90147-7. [DOI] [PubMed] [Google Scholar]
  12. Johnson R. C., Manasek F. J., Vinson W. C., Seyer J. M. The biochemical and ultrastructural demonstration of collagen during early heart development. Dev Biol. 1974 Feb;36(2):252–271. doi: 10.1016/0012-1606(74)90049-9. [DOI] [PubMed] [Google Scholar]
  13. Kinsella M. G., Fitzharris T. P. Control of cell migration in atrioventricular pads during chick early heart development: analysis of cushion tissue migration in vitro. Dev Biol. 1982 May;91(1):1–10. doi: 10.1016/0012-1606(82)90002-1. [DOI] [PubMed] [Google Scholar]
  14. Kinsella M. G., Fitzharris T. P. Origin of cushion tissue in the developing chick heart: cinematographic recordings of in situ formation. Science. 1980 Mar 21;207(4437):1359–1360. doi: 10.1126/science.7355294. [DOI] [PubMed] [Google Scholar]
  15. Klebe R. J. Isolation of a collagen-dependent cell attachment factor. Nature. 1974 Jul 19;250(463):248–251. doi: 10.1038/250248a0. [DOI] [PubMed] [Google Scholar]
  16. Knox P., Wells P. Cell adhesion and proteoglycans. I. The effect of exogenous proteoglycans on the attachment of chick embryo fibroblasts to tissue culture plastic and collagen. J Cell Sci. 1979 Dec;40:77–88. doi: 10.1242/jcs.40.1.77. [DOI] [PubMed] [Google Scholar]
  17. Lark M. W., Culp L. A. Selective solubilization of hyaluronic acid from fibroblast substratum adhesion sites. J Biol Chem. 1982 Dec 10;257(23):14073–14080. [PubMed] [Google Scholar]
  18. Laterra J., Ansbacher R., Culp L. A. Glycosaminoglycans that bind cold-insoluble globulin in cell-substratum adhesion sites of murine fibroblasts. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6662–6666. doi: 10.1073/pnas.77.11.6662. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Laterra J., Norton E. K., Izzard C. S., Culp L. A. Contact formation by fibroblasts adhering to heparan sulfate-binding substrata (fibronectin or platelet factor 4). Exp Cell Res. 1983 Jun;146(1):15–27. doi: 10.1016/0014-4827(83)90320-8. [DOI] [PubMed] [Google Scholar]
  20. Lindahl U., Bäckström G., Jansson L., Hallén A. Biosynthesis of heparin. II. Formation of sulfamino groups. J Biol Chem. 1973 Oct 25;248(20):7234–7241. [PubMed] [Google Scholar]
  21. Linker A., Hovingh P. The uses of degradative enzymes as tools for identification and structural analysis of glycosaminoglycans. Fed Proc. 1977 Jan;36(1):43–46. [PubMed] [Google Scholar]
  22. Manasek F. J., Reid M., Vinson W., Seyer J., Johnson R. Glycosaminoglycan synthesis by the early embryonic chick heart. Dev Biol. 1973 Dec;35(2):332–348. doi: 10.1016/0012-1606(73)90028-6. [DOI] [PubMed] [Google Scholar]
  23. Manasek F. J. Structural glycoproteins of the embryonic cardiac extracellular matrix. J Mol Cell Cardiol. 1977 Jun;9(6):425–439. doi: 10.1016/s0022-2828(77)80024-2. [DOI] [PubMed] [Google Scholar]
  24. Markwald R. R., Fitzharris T. P., Bank H., Bernanke D. H. Structural analyses on the matrical organization of glycosaminoglycans in developing endocardial cushions. Dev Biol. 1978 Feb;62(2):292–316. doi: 10.1016/0012-1606(78)90218-x. [DOI] [PubMed] [Google Scholar]
  25. Markwald R. R., Fitzharris T. P., Bolender D. L., Bernanke D. H. Sturctural analysis of cell:matrix association during the morphogenesis of atrioventricular cushion tissue. Dev Biol. 1979 Apr;69(2):634–654. doi: 10.1016/0012-1606(79)90317-8. [DOI] [PubMed] [Google Scholar]
  26. Markwald R. R., Fitzharris T. P., Smith W. N. Sturctural analysis of endocardial cytodifferentiation. Dev Biol. 1975 Jan;42(1):160–180. doi: 10.1016/0012-1606(75)90321-8. [DOI] [PubMed] [Google Scholar]
  27. Markwald R. R., Funderburg F. M. Use of 6-diazo-5-oxo-L-norleucine to study interaction between myocardial glycoconjugate secretion and endothelial activation in the early embryonic chick heart. Dev Biol. 1983 Oct;99(2):395–407. doi: 10.1016/0012-1606(83)90289-0. [DOI] [PubMed] [Google Scholar]
  28. Markwald R. R., Krook J. M., Kitten G. T., Runyan R. B. Endocardial cushion tissue development: structural analyses on the attachment of extracellular matrix to migrating mesenchymal cell surfaces. Scan Electron Microsc. 1981;(Pt 2):261–274. [PubMed] [Google Scholar]
  29. Newgreen D. F., Scheel M., Kastner V. Morphogenesis of sclerotome and neural crest in avian embryos. In vivo and in vitro studies on the role of notochordal extracellular material. Cell Tissue Res. 1986;244(2):299–313. doi: 10.1007/BF00219205. [DOI] [PubMed] [Google Scholar]
  30. Pytela R., Pierschbacher M. D., Ruoslahti E. Identification and isolation of a 140 kd cell surface glycoprotein with properties expected of a fibronectin receptor. Cell. 1985 Jan;40(1):191–198. doi: 10.1016/0092-8674(85)90322-8. [DOI] [PubMed] [Google Scholar]
  31. Rich A. M., Pearlstein E., Weissmann G., Hoffstein S. T. Cartilage proteoglycans inhibit fibronectin-mediated adhesion. Nature. 1981 Sep 17;293(5829):224–226. doi: 10.1038/293224a0. [DOI] [PubMed] [Google Scholar]
  32. Rollins B. J., Culp L. A. Preliminary characterization of the proteoglycans in the substrate adhesion sites of normal and virus-transformed murine cells. Biochemistry. 1979 Dec 11;18(25):5621–5629. doi: 10.1021/bi00592a016. [DOI] [PubMed] [Google Scholar]
  33. Runyan R. B., Markwald R. R. Invasion of mesenchyme into three-dimensional collagen gels: a regional and temporal analysis of interaction in embryonic heart tissue. Dev Biol. 1983 Jan;95(1):108–114. doi: 10.1016/0012-1606(83)90010-6. [DOI] [PubMed] [Google Scholar]
  34. Saito H., Yamagata T., Suzuki S. Enzymatic methods for the determination of small quantities of isomeric chondroitin sulfates. J Biol Chem. 1968 Apr 10;243(7):1536–1542. [PubMed] [Google Scholar]
  35. Smith G. N., Jr, Williams J. M., Brandt K. D. Interaction of proteoglycans with the pericellular (1 alpha, 2 alpha, 3 alpha) collagens of cartilage. J Biol Chem. 1985 Sep 5;260(19):10761–10767. [PubMed] [Google Scholar]
  36. Tucker R. P., Erickson C. A. Morphology and behavior of quail neural crest cells in artificial three-dimensional extracellular matrices. Dev Biol. 1984 Aug;104(2):390–405. doi: 10.1016/0012-1606(84)90094-0. [DOI] [PubMed] [Google Scholar]
  37. Turley E. A., Erickson C. A., Tucker R. P. The retention and ultrastructural appearances of various extracellular matrix molecules incorporated into three-dimensional hydrated collagen lattices. Dev Biol. 1985 Jun;109(2):347–369. doi: 10.1016/0012-1606(85)90461-0. [DOI] [PubMed] [Google Scholar]
  38. Vogel K. G., Kendall V. F. Cell-surface glycosaminoglycans: turnover in cultured human embryo fibroblasts (IMR-90). J Cell Physiol. 1980 Jun;103(3):475–487. doi: 10.1002/jcp.1041030313. [DOI] [PubMed] [Google Scholar]
  39. Wasserman L., Ber A., Allalouf D. Use of thin-layer chromatography in the separation of disaccharides resulting from digestion of chondroitin sulphates with chondroitinases. J Chromatogr. 1977 Jun 11;136(2):342–347. doi: 10.1016/s0021-9673(00)86291-3. [DOI] [PubMed] [Google Scholar]
  40. Yamada K. M. Cell surface interactions with extracellular materials. Annu Rev Biochem. 1983;52:761–799. doi: 10.1146/annurev.bi.52.070183.003553. [DOI] [PubMed] [Google Scholar]

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

RESOURCES