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. 1983 Apr 1;96(4):1108–1116. doi: 10.1083/jcb.96.4.1108

Degradation of extracellular matrix by mouse trophoblast outgrowths: a model for implantation

RH Glass, J Aggeler, A Spindle, RA Pederson, Z Werb
PMCID: PMC2112312  PMID: 6339525

Abstract

During implantation the embryo attaches to the endometrial surface and trophoblast traverses the uterine epithelium, anchoring in the uterine connective tissue. To determine whether trophoblast can facilitate invasion of the uterus by degrading components of normal uterine extracellular matrix, mouse blastocysts were cultured on a radio-labeled extracellular matrix that contained glycoproteins, elastin, and collagen. The embryos attached to the matrix, and trophoblast spread over the surface. Starting on day 5 of culture there was a release of labeled peptides into the medium. The radioactive peptides released from the matrix by the embryos had molecular weights ranging from more than 25,000 to more than 200. By day 7 there were areas where individual trophoblast cells had separated from one another, revealing the underlying substratum that was cleared of matrix. When trophoblast cells were lysed with NH(4)OH on day 8, it was apparent that the area underneath the trophoblast outgrowth had been cleared of matrix. Scanning electron microscopy and time-lapse cinemicrography confirmed that the digestion of matrix was highly localized, taking place only underneath the trophoblast, with no evidence of digestion of the matrix beyond the periphery of the trophoblast outgrowth. The sharp boundaries of degredation observed may be due to localized proteinase secretion by trophoblast, to membrane proteinases on the surface of trophoblast, or to endocytosis. Digestion of the matrix was not dependent on plasminogen, thus ruling out a role for plasminogen activator. Digestion was not inhibited by a variety of hormones and inhibitors, including progesterone, 17β-estradiol, leupeptin, EDTA, colchicine, NH(4)Cl, or ε-aminocaproic acid. This system of culturing embryos on extracellular matrix may be useful in determining the processes that regulate trophoblast migration and invasion into the maternal tissues during implantation.0

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Selected References

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

  1. Aggeler J., Kapp L. N., Tseng S. C., Werb Z. Regulation of protein secretion in Chinese hamster ovary cells by cell cycle position and cell density. Plasminogen activator, procollagen fibronectin. Exp Cell Res. 1982 Jun;139(2):275–283. doi: 10.1016/0014-4827(82)90252-x. [DOI] [PubMed] [Google Scholar]
  2. Chapman H. A., Jr, Vavrin Z., Hibbs J. B., Jr Macrophage fibrinolytic activity: identification of two pathways of plasmin formation by intact cells and of a plasminogen activator inhibitor. Cell. 1982 Mar;28(3):653–662. doi: 10.1016/0092-8674(82)90220-3. [DOI] [PubMed] [Google Scholar]
  3. Dabich D., Andary T. J. Prevention of blastocyst implantation in mice with proteinase inhibitors. Fertil Steril. 1974 Nov;25(11):954–957. [PubMed] [Google Scholar]
  4. 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]
  5. Dubin N. H., Cummings D. B., Blake D. A., King T. M. Effect of epsilon amino caproic acid, a fibrinolytic inhibitor, on implantation and fetal viability in the rat. Biol Reprod. 1980 Oct;23(3):553–557. doi: 10.1095/biolreprod23.3.553. [DOI] [PubMed] [Google Scholar]
  6. Glass R. H., Spindle A. I., Maglio M., Pedersen R. A. The free surface of mouse trophoblast in culture is non-adhesive for other cells. J Reprod Fertil. 1980 Jul;59(2):403–407. doi: 10.1530/jrf.0.0590403. [DOI] [PubMed] [Google Scholar]
  7. Glass R. H., Spindle A. I., Pedersen R. A. Mouse embryo attachment to substratum and interaction of trophoblast with cultured cells. J Exp Zool. 1979 Jun;208(3):327–336. doi: 10.1002/jez.1402080309. [DOI] [PubMed] [Google Scholar]
  8. Gwatkin R. B. Defined media and development of mammalian eggs in vitro. Ann N Y Acad Sci. 1966 Oct 7;139(1):79–90. doi: 10.1111/j.1749-6632.1966.tb41186.x. [DOI] [PubMed] [Google Scholar]
  9. Hsu Y. C. Differentiation in vitro of mouse embryos beyond the implantation stage. Nature. 1972 Sep 22;239(5369):200–202. doi: 10.1038/239200a0. [DOI] [PubMed] [Google Scholar]
  10. Johnson K. J., Varani J. Substrate hydrolysis by immune complex-activated neutrophils: effect of physical presentation of complexes and protease inhibitors. J Immunol. 1981 Nov;127(5):1875–1879. [PubMed] [Google Scholar]
  11. Jones P. A., DeClerck Y. A. Destruction of extracellular matrices containing glycoproteins, elastin, and collagen by metastatic human tumor cells. Cancer Res. 1980 Sep;40(9):3222–3227. [PubMed] [Google Scholar]
  12. Jones P. A., Scott-Burden T. Activated macrophages digest the extracellular matrix proteins produced by cultured cells. Biochem Biophys Res Commun. 1979 Jan 15;86(1):71–77. doi: 10.1016/0006-291x(79)90383-8. [DOI] [PubMed] [Google Scholar]
  13. Jones P. A., Scott-Burden T., Gevers W. Glycoprotein, elastin, and collagen secretion by rat smooth muscle cells. Proc Natl Acad Sci U S A. 1979 Jan;76(1):353–357. doi: 10.1073/pnas.76.1.353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kubo H., Katayama S., Amano H., Spindle A. I. Plasminogen activator activity in mouse embryos cultured on decidual cell monolayers. Nihon Sanka Fujinka Gakkai Zasshi. 1982 Jun;34(6):801–808. [PubMed] [Google Scholar]
  15. Kubo H., Spindle A., Pedersen R. A. Inhibition of mouse blastocyst attachment and outgrowth by protease inhibitors. J Exp Zool. 1981 Jun;216(3):445–451. doi: 10.1002/jez.1402160313. [DOI] [PubMed] [Google Scholar]
  16. Muńoz M. L., Calderón J., Rojkind M. The collagenase of Entamoeba histolytica. J Exp Med. 1982 Jan 1;155(1):42–51. doi: 10.1084/jem.155.1.42. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Russo R. G., Liotta L. A., Thorgeirsson U., Brundage R., Schiffmann E. Polymorphonuclear leukocyte migration through human amnion membrane. J Cell Biol. 1981 Nov;91(2 Pt 1):459–467. doi: 10.1083/jcb.91.2.459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Schlafke S., Enders A. C. Cellular basis of interaction between trophoblast and uterus at implantation. Biol Reprod. 1975 Feb;12(1):41–65. doi: 10.1095/biolreprod12.1.41. [DOI] [PubMed] [Google Scholar]
  19. Schor S. L., Allen T. D., Harrison C. J. Cell migration through three-dimensional gels of native collagen fibres: collagenolytic activity is not required for the migration of two permanent cell lines. J Cell Sci. 1980 Dec;46:171–186. doi: 10.1242/jcs.46.1.171. [DOI] [PubMed] [Google Scholar]
  20. Sherman M. I., Salomon D. S. The relationships between the early mouse embryo and its environment. Symp Soc Dev Biol. 1975;(33):277–309. doi: 10.1016/b978-0-12-612979-3.50019-4. [DOI] [PubMed] [Google Scholar]
  21. Sobel J. S., Cooke R., Pedersen R. A. Distribution of actin and myosin in mouse trophoblast: correlation with changes in invasiveness during development in vitro. Dev Biol. 1980 Aug;78(2):365–379. doi: 10.1016/0012-1606(80)90341-3. [DOI] [PubMed] [Google Scholar]
  22. Spindle A. I., Pedersen R. A. Hatching, attachment, and outgrowth of mouse blastocysts in vitro: fixed nitrogen requirements. J Exp Zool. 1973 Dec;186(3):305–318. doi: 10.1002/jez.1401860308. [DOI] [PubMed] [Google Scholar]
  23. Strickland S., Reich E., Sherman M. I. Plasminogen activator in early embryogenesis: enzyme production by trophoblast and parietal endoderm. Cell. 1976 Oct;9(2):231–240. doi: 10.1016/0092-8674(76)90114-8. [DOI] [PubMed] [Google Scholar]
  24. Unkeless J. C., Gordon S., Reich E. Secretion of plasminogen activator by stimulated macrophages. J Exp Med. 1974 Apr 1;139(4):834–850. doi: 10.1084/jem.139.4.834. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Werb Z., Bainton D. F., Jones P. A. Degradation of connective tissue matrices by macrophages. III. Morphological and biochemical studies on extracellular, pericellular, and intracellular events in matrix proteolysis by macrophages in culture. J Exp Med. 1980 Dec 1;152(6):1537–1553. doi: 10.1084/jem.152.6.1537. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Werb Z., Banda M. J., Jones P. A. Degradation of connective tissue matrices by macrophages. I. Proteolysis of elastin, glycoproteins, and collagen by proteinases isolated from macrophages. J Exp Med. 1980 Nov 1;152(5):1340–1357. doi: 10.1084/jem.152.5.1340. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Wiley L. M., Pedersen R. A. Morphology of mouse egg cylinder development in vitro: a light and electron microscopic study. J Exp Zool. 1977 Jun;200(3):389–402. doi: 10.1002/jez.1402000309. [DOI] [PubMed] [Google Scholar]
  28. Wilson I. B., Jenkinson E. J. Blastocyst differentiation in vitro. J Reprod Fertil. 1974 Jul;39(1):243–249. doi: 10.1530/jrf.0.0390243. [DOI] [PubMed] [Google Scholar]

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