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. 1990 Apr 1;110(4):1405–1415. doi: 10.1083/jcb.110.4.1405

Expression of extracellular matrix components is regulated by substratum

PMCID: PMC2116068  PMID: 2182652

Abstract

Reconstituted basement membranes and extracellular matrices have been demonstrated to affect, positively and dramatically, the production of milk proteins in cultured mammary epithelial cells. Here we show that both the expression and the deposition of extracellular matrix components themselves are regulated by substratum. The steady-state levels of the laminin, type IV collagen, and fibronectin mRNAs in mammary epithelial cells cultured on plastic dishes and on type I collagen gels have been examined, as has the ability of these cells to synthesize, secrete, and deposit laminin and other, extracellular matrix proteins. We demonstrate de novo synthesis of a basement membrane by cells cultured on type I collagen gels which have been floated into the medium. Expression of the mRNA and proteins of basement membranes, however, are quite low in these cultures. In contrast, the levels of laminin, type IV collagen, and fibronectin mRNAs are highest in cells cultured on plastic surfaces, where no basement membrane is deposited. It is suggested that the interaction between epithelial cells and both basement membrane and stromally derived matrices exerts a negative influence on the expression of mRNA for extracellular matrix components. In addition, we show that the capacity for lactational differentiation correlates with conditions that favor the deposition of a continuous basement membrane, and argue that the interaction between specialized epithelial cells and stroma enables them to create their own microenvironment for accurate signal transduction and phenotypic function.

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Selected References

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  1. Barcellos-Hoff M. H., Aggeler J., Ram T. G., Bissell M. J. Functional differentiation and alveolar morphogenesis of primary mammary cultures on reconstituted basement membrane. Development. 1989 Feb;105(2):223–235. doi: 10.1242/dev.105.2.223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barlow D. P., Green N. M., Kurkinen M., Hogan B. L. Sequencing of laminin B chain cDNAs reveals C-terminal regions of coiled-coil alpha-helix. EMBO J. 1984 Oct;3(10):2355–2362. doi: 10.1002/j.1460-2075.1984.tb02140.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bissell D. M., Arenson D. M., Maher J. J., Roll F. J. Support of cultured hepatocytes by a laminin-rich gel. Evidence for a functionally significant subendothelial matrix in normal rat liver. J Clin Invest. 1987 Mar;79(3):801–812. doi: 10.1172/JCI112887. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bissell M. J., Hall H. G., Parry G. How does the extracellular matrix direct gene expression? J Theor Biol. 1982 Nov 7;99(1):31–68. doi: 10.1016/0022-5193(82)90388-5. [DOI] [PubMed] [Google Scholar]
  5. Blum J. L., Wicha M. S. Role of the cytoskeleton in laminin induced mammary gene expression. J Cell Physiol. 1988 Apr;135(1):13–22. doi: 10.1002/jcp.1041350103. [DOI] [PubMed] [Google Scholar]
  6. Blum J. L., Zeigler M. E., Wicha M. S. Regulation of rat mammary gene expression by extracellular matrix components. Exp Cell Res. 1987 Dec;173(2):322–340. doi: 10.1016/0014-4827(87)90274-6. [DOI] [PubMed] [Google Scholar]
  7. Bonner W. M. Autoradiograms: 35S and 32P. Methods Enzymol. 1987;152:55–61. doi: 10.1016/0076-6879(87)52010-9. [DOI] [PubMed] [Google Scholar]
  8. Burwen S. J., Pitelka D. R. Secretory function of lactating mouse mammary epithelial cells cultured on collagen gels. Exp Cell Res. 1980 Apr;126(2):249–262. doi: 10.1016/0014-4827(80)90263-3. [DOI] [PubMed] [Google Scholar]
  9. Caplan M. J., Stow J. L., Newman A. P., Madri J., Anderson H. C., Farquhar M. G., Palade G. E., Jamieson J. D. Dependence on pH of polarized sorting of secreted proteins. Nature. 1987 Oct 15;329(6140):632–635. doi: 10.1038/329632a0. [DOI] [PubMed] [Google Scholar]
  10. Carey D. J., Todd M. S., Rafferty C. M. Schwann cell myelination: induction by exogenous basement membrane-like extracellular matrix. J Cell Biol. 1986 Jun;102(6):2254–2263. doi: 10.1083/jcb.102.6.2254. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Chen L. H., Bissell M. J. A novel regulatory mechanism for whey acidic protein gene expression. Cell Regul. 1989 Nov;1(1):45–54. doi: 10.1091/mbc.1.1.45. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Chen L. H., Bissell M. J. Transferrin mRNA level in the mouse mammary gland is regulated by pregnancy and extracellular matrix. J Biol Chem. 1987 Dec 25;262(36):17247–17250. [PubMed] [Google Scholar]
  13. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  14. David G., Bernfield M. Type I collagen reduces the degradation of basal lamina proteoglycan by mammary epithelial cells. J Cell Biol. 1981 Oct;91(1):281–286. doi: 10.1083/jcb.91.1.281. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. David G., Nusgens B., van der Schueren B., van Cauwenberge D., van den Berghe H., Lapière C. Collagen metabolism and basement membrane formation in cultures of mouse mammary epithelial cells. Induction of 'assembly' on fibrillar type I collagen substrata. Exp Cell Res. 1987 Jun;170(2):402–416. doi: 10.1016/0014-4827(87)90316-8. [DOI] [PubMed] [Google Scholar]
  16. Eisenstein R. S., Rosen J. M. Both cell substratum regulation and hormonal regulation of milk protein gene expression are exerted primarily at the posttranscriptional level. Mol Cell Biol. 1988 Aug;8(8):3183–3190. doi: 10.1128/mcb.8.8.3183. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Emerman J. T., Enami J., Pitelka D. R., Nandi S. Hormonal effects on intracellular and secreted casein in cultures of mouse mammary epithelial cells on floating collagen membranes. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4466–4470. doi: 10.1073/pnas.74.10.4466. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Emerman J. T., Pitelka D. R. Maintenance and induction of morphological differentiation in dissociated mammary epithelium on floating collagen membranes. In Vitro. 1977 May;13(5):316–328. doi: 10.1007/BF02616178. [DOI] [PubMed] [Google Scholar]
  19. Feinberg A. P., Vogelstein B. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984 Feb;137(1):266–267. doi: 10.1016/0003-2697(84)90381-6. [DOI] [PubMed] [Google Scholar]
  20. Glasser S. R., Julian J., Decker G. L., Tang J. P., Carson D. D. Development of morphological and functional polarity in primary cultures of immature rat uterine epithelial cells. J Cell Biol. 1988 Dec;107(6 Pt 1):2409–2423. doi: 10.1083/jcb.107.6.2409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Grant D. S., Leblond C. P., Kleinman H. K., Inoue S., Hassell J. R. The incubation of laminin, collagen IV, and heparan sulfate proteoglycan at 35 degrees C yields basement membrane-like structures. J Cell Biol. 1989 Apr;108(4):1567–1574. doi: 10.1083/jcb.108.4.1567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hadley M. A., Byers S. W., Suárez-Quian C. A., Djakiew D., Dym M. In vitro models of differentiated Sertoli cell structure and function. In Vitro Cell Dev Biol. 1988 Jun;24(6):550–557. doi: 10.1007/BF02629090. [DOI] [PubMed] [Google Scholar]
  23. Hadley M. A., Byers S. W., Suárez-Quian C. A., Kleinman H. K., Dym M. Extracellular matrix regulates Sertoli cell differentiation, testicular cord formation, and germ cell development in vitro. J Cell Biol. 1985 Oct;101(4):1511–1522. doi: 10.1083/jcb.101.4.1511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Haeuptle M. T., Suard Y. L., Bogenmann E., Reggio H., Racine L., Kraehenbuhl J. P. Effect of cell shape change on the function and differentiation of rabbit mammary cells in culture. J Cell Biol. 1983 May;96(5):1425–1434. doi: 10.1083/jcb.96.5.1425. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Hedin U., Bottger B. A., Forsberg E., Johansson S., Thyberg J. Diverse effects of fibronectin and laminin on phenotypic properties of cultured arterial smooth muscle cells. J Cell Biol. 1988 Jul;107(1):307–319. doi: 10.1083/jcb.107.1.307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Ignotz R. A., Massagué J. Transforming growth factor-beta stimulates the expression of fibronectin and collagen and their incorporation into the extracellular matrix. J Biol Chem. 1986 Mar 25;261(9):4337–4345. [PubMed] [Google Scholar]
  27. Ingber D. E., Folkman J. How does extracellular matrix control capillary morphogenesis? Cell. 1989 Sep 8;58(5):803–805. doi: 10.1016/0092-8674(89)90928-8. [DOI] [PubMed] [Google Scholar]
  28. Kimata K., Sakakura T., Inaguma Y., Kato M., Nishizuka Y. Participation of two different mesenchymes in the developing mouse mammary gland: synthesis of basement membrane components by fat pad precursor cells. J Embryol Exp Morphol. 1985 Oct;89:243–257. [PubMed] [Google Scholar]
  29. Kleinman H. K., McGarvey M. L., Hassell J. R., Martin G. R. Formation of a supramolecular complex is involved in the reconstitution of basement membrane components. Biochemistry. 1983 Oct 11;22(21):4969–4974. doi: 10.1021/bi00290a014. [DOI] [PubMed] [Google Scholar]
  30. Kubota Y., Kleinman H. K., Martin G. R., Lawley T. J. Role of laminin and basement membrane in the morphological differentiation of human endothelial cells into capillary-like structures. J Cell Biol. 1988 Oct;107(4):1589–1598. doi: 10.1083/jcb.107.4.1589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Kurkinen M., Barlow D. P., Helfman D. M., Williams J. G., Hogan B. L. Isolation of cDNA clones for basal lamina components: type IV procollagen. Nucleic Acids Res. 1983 Sep 24;11(18):6199–6209. doi: 10.1093/nar/11.18.6199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Lane E. B. Monoclonal antibodies provide specific intramolecular markers for the study of epithelial tonofilament organization. J Cell Biol. 1982 Mar;92(3):665–673. doi: 10.1083/jcb.92.3.665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Lee E. Y., Lee W. H., Kaetzel C. S., Parry G., Bissell M. J. Interaction of mouse mammary epithelial cells with collagen substrata: regulation of casein gene expression and secretion. Proc Natl Acad Sci U S A. 1985 Mar;82(5):1419–1423. doi: 10.1073/pnas.82.5.1419. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Lee E. Y., Parry G., Bissell M. J. Modulation of secreted proteins of mouse mammary epithelial cells by the collagenous substrata. J Cell Biol. 1984 Jan;98(1):146–155. doi: 10.1083/jcb.98.1.146. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Li M. L., Aggeler J., Farson D. A., Hatier C., Hassell J., Bissell M. J. Influence of a reconstituted basement membrane and its components on casein gene expression and secretion in mouse mammary epithelial cells. Proc Natl Acad Sci U S A. 1987 Jan;84(1):136–140. doi: 10.1073/pnas.84.1.136. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Madri J. A., Pratt B. M., Tucker A. M. Phenotypic modulation of endothelial cells by transforming growth factor-beta depends upon the composition and organization of the extracellular matrix. J Cell Biol. 1988 Apr;106(4):1375–1384. doi: 10.1083/jcb.106.4.1375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Nickerson S. C., Akers R. M. Biochemical and ultrastructural aspects of milk synthesis and secretion. Int J Biochem. 1984;16(8):855–865. doi: 10.1016/0020-711x(84)90144-7. [DOI] [PubMed] [Google Scholar]
  38. Parry G., Cullen B., Kaetzel C. S., Kramer R., Moss L. Regulation of differentiation and polarized secretion in mammary epithelial cells maintained in culture: extracellular matrix and membrane polarity influences. J Cell Biol. 1987 Nov;105(5):2043–2051. doi: 10.1083/jcb.105.5.2043. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Parry G., Lee E. Y., Farson D., Koval M., Bissell M. J. Collagenous substrata regulate the nature and distribution of glycosaminoglycans produced by differentiated cultures of mouse mammary epithelial cells. Exp Cell Res. 1985 Feb;156(2):487–499. doi: 10.1016/0014-4827(85)90556-7. [DOI] [PubMed] [Google Scholar]
  40. Peters B. P., Hartle R. J., Krzesicki R. F., Kroll T. G., Perini F., Balun J. E., Goldstein I. J., Ruddon R. W. The biosynthesis, processing, and secretion of laminin by human choriocarcinoma cells. J Biol Chem. 1985 Nov 25;260(27):14732–14742. [PubMed] [Google Scholar]
  41. Reichmann E., Ball R., Groner B., Friis R. R. New mammary epithelial and fibroblastic cell clones in coculture form structures competent to differentiate functionally. J Cell Biol. 1989 Mar;108(3):1127–1138. doi: 10.1083/jcb.108.3.1127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Rodriguez-Boulan E., Nelson W. J. Morphogenesis of the polarized epithelial cell phenotype. Science. 1989 Aug 18;245(4919):718–725. doi: 10.1126/science.2672330. [DOI] [PubMed] [Google Scholar]
  43. Schuetz E. G., Li D., Omiecinski C. J., Muller-Eberhard U., Kleinman H. K., Elswick B., Guzelian P. S. Regulation of gene expression in adult rat hepatocytes cultured on a basement membrane matrix. J Cell Physiol. 1988 Mar;134(3):309–323. doi: 10.1002/jcp.1041340302. [DOI] [PubMed] [Google Scholar]
  44. Schwarzbauer J. E., Tamkun J. W., Lemischka I. R., Hynes R. O. Three different fibronectin mRNAs arise by alternative splicing within the coding region. Cell. 1983 Dec;35(2 Pt 1):421–431. doi: 10.1016/0092-8674(83)90175-7. [DOI] [PubMed] [Google Scholar]
  45. Servely J. L., Geuens G. M., Martel P., Houdebine L. M., de Brabander M. Effect of tubulozole, a new synthetic microtubule inhibitor, on the induction of casein gene expression by prolactin. Biol Cell. 1987;59(2):121–127. doi: 10.1111/j.1768-322x.1987.tb00522.x. [DOI] [PubMed] [Google Scholar]
  46. Shannon J. M., Mason R. J., Jennings S. D. Functional differentiation of alveolar type II epithelial cells in vitro: effects of cell shape, cell-matrix interactions and cell-cell interactions. Biochim Biophys Acta. 1987 Nov 12;931(2):143–156. doi: 10.1016/0167-4889(87)90200-x. [DOI] [PubMed] [Google Scholar]
  47. Shannon J. M., Pitelka D. R. The influence of cell shape on the induction of functional differentiation in mouse mammary cells in vitro. In Vitro. 1981 Nov;17(11):1016–1028. doi: 10.1007/BF02618428. [DOI] [PubMed] [Google Scholar]
  48. Sugrue S. P., Hay E. D. The identification of extracellular matrix (ECM) binding sites on the basal surface of embryonic corneal epithelium and the effect of ECM binding on epithelial collagen production. J Cell Biol. 1986 May;102(5):1907–1916. doi: 10.1083/jcb.102.5.1907. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Unemori E. N., Bouhana K. S., Werb Z. Vectorial secretion of extracellular matrix proteins, matrix-degrading proteinases, and tissue inhibitor of metalloproteinases by endothelial cells. J Biol Chem. 1990 Jan 5;265(1):445–451. [PubMed] [Google Scholar]
  50. Warburton M. J., Kimbell R., Rudland P. S., Ferns S. A., Barraclough R. Control of type IV collagen production in rat mammary epithelial and myoepithelial-like cells. J Cell Physiol. 1986 Jul;128(1):76–84. doi: 10.1002/jcp.1041280113. [DOI] [PubMed] [Google Scholar]
  51. Wiens D., Park C. S., Stockdale F. E. Milk protein expression and ductal morphogenesis in the mammary gland in vitro: hormone-dependent and -independent phases of adipocyte-mammary epithelial cell interaction. Dev Biol. 1987 Mar;120(1):245–258. doi: 10.1016/0012-1606(87)90122-9. [DOI] [PubMed] [Google Scholar]
  52. Wrana J. L., Maeno M., Hawrylyshyn B., Yao K. L., Domenicucci C., Sodek J. Differential effects of transforming growth factor-beta on the synthesis of extracellular matrix proteins by normal fetal rat calvarial bone cell populations. J Cell Biol. 1988 Mar;106(3):915–924. doi: 10.1083/jcb.106.3.915. [DOI] [PMC free article] [PubMed] [Google Scholar]

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