Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1990 Mar 1;110(3):849–857. doi: 10.1083/jcb.110.3.849

Type IV collagen mRNA accumulates in the mesenchymal compartment at early stages of murine developing intestine

PMCID: PMC2116038  PMID: 2307711

Abstract

The expression of type IV collagen mRNA during mouse intestinal morphogenesis was examined by in situ hybridization using a cDNA probe corresponding to mRNA for alpha 1 (IV) chain. Type IV collagen mRNA is detected in the embryonic mesenchymal cells at early stages of development (12 d of gestation). A segregation of mesenchymal cells expressing high levels of type IV collagen mRNA in close vicinity of the epithelium occurs just before villus formation. During villus outgrowth, type IV collagen mRNA, still confined to mesenchyme-derived tissues, is progressively restricted to the mucosal connective tissue (the lamina propria) and to a lesser extent to the muscular layers. In the adult, the amount of messenger is quite low as compared to the level found in the developing intestine and the in situ hybridization signal, indistinguishable from the background, is uniform throughout the whole intestinal wall. At all developmental stages no detectable specific hybridization signal is virtually observed over the epithelium cell layer. These results show that high amounts of the type IV collagen messenger are detected during phases of intensive morphogenetic events. Furthermore, they reinforce the notion already gained previously (Simon-Assmann et al. 1988) that the mesenchymal compartment is the principal endogenous source of type IV collagen. They also indicate that the continuous migration of epithelial cells along the basement membrane of intestinal villi in the mature organ is not accompanied by a significant remodeling of the collagen IV network.

Full Text

The Full Text of this article is available as a PDF (3.5 MB).

Selected References

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

  1. Aufderheide E., Ekblom P. Tenascin during gut development: appearance in the mesenchyme, shift in molecular forms, and dependence on epithelial-mesenchymal interactions. J Cell Biol. 1988 Dec;107(6 Pt 1):2341–2349. doi: 10.1083/jcb.107.6.2341. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Auffray C., Rougeon F. Purification of mouse immunoglobulin heavy-chain messenger RNAs from total myeloma tumor RNA. Eur J Biochem. 1980 Jun;107(2):303–314. doi: 10.1111/j.1432-1033.1980.tb06030.x. [DOI] [PubMed] [Google Scholar]
  3. Berger C. N. In situ hybridization of immunoglobulin-specific RNA in single cells of the B lymphocyte lineage with radiolabelled DNA probes. EMBO J. 1986 Jan;5(1):85–93. doi: 10.1002/j.1460-2075.1986.tb04181.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dente L., Cesareni G., Cortese R. pEMBL: a new family of single stranded plasmids. Nucleic Acids Res. 1983 Mar 25;11(6):1645–1655. doi: 10.1093/nar/11.6.1645. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Devlin C. J., Brickell P. M., Taylor E. R., Hornbruch A., Craig R. K., Wolpert L. In situ hybridization reveals differential spatial distribution of mRNAs for type I and type II collagen in the chick limb bud. Development. 1988 May;103(1):111–118. doi: 10.1242/dev.103.1.111. [DOI] [PubMed] [Google Scholar]
  6. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  7. Hafen E., Levine M., Garber R. L., Gehring W. J. An improved in situ hybridization method for the detection of cellular RNAs in Drosophila tissue sections and its application for localizing transcripts of the homeotic Antennapedia gene complex. EMBO J. 1983;2(4):617–623. doi: 10.1002/j.1460-2075.1983.tb01472.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Haffen K., Kedinger M., Simon-Assmann P. Mesenchyme-dependent differentiation of epithelial progenitor cells in the gut. J Pediatr Gastroenterol Nutr. 1987 Jan-Feb;6(1):14–23. doi: 10.1097/00005176-198701000-00005. [DOI] [PubMed] [Google Scholar]
  9. Hayashi M., Ninomiya Y., Parsons J., Hayashi K., Olsen B. R., Trelstad R. L. Differential localization of mRNAs of collagen types I and II in chick fibroblasts, chondrocytes, and corneal cells by in situ hybridization using cDNA probes. J Cell Biol. 1986 Jun;102(6):2302–2309. doi: 10.1083/jcb.102.6.2302. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hayashi S., Gillam I. C., Delaney A. D., Tener G. M. Acetylation of chromosome squashes of Drosophila melanogaster decreases the background in autoradiographs from hybridization with [125I]-labeled RNA. J Histochem Cytochem. 1978 Aug;26(8):677–679. doi: 10.1177/26.8.99471. [DOI] [PubMed] [Google Scholar]
  11. Kühl U., Ocalan M., Timpl R., Mayne R., Hay E., von der Mark K. Role of muscle fibroblasts in the deposition of type-IV collagen in the basal lamina of myotubes. Differentiation. 1984;28(2):164–172. doi: 10.1111/j.1432-0436.1984.tb00279.x. [DOI] [PubMed] [Google Scholar]
  12. Kühl U., Timpl R., von der Mark K. Synthesis of type IV collagen and laminin in cultures of skeletal muscle cells and their assembly on the surface of myotubes. Dev Biol. 1982 Oct;93(2):344–354. doi: 10.1016/0012-1606(82)90122-1. [DOI] [PubMed] [Google Scholar]
  13. Lehrach H., Diamond D., Wozney J. M., Boedtker H. RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry. 1977 Oct 18;16(21):4743–4751. doi: 10.1021/bi00640a033. [DOI] [PubMed] [Google Scholar]
  14. Mirre C., Cecchini J. P., Le Parco Y., Knibiehler B. De novo expression of a type IV collagen gene in Drosophila embryos is restricted to mesodermal derivatives and occurs at germ band shortening. Development. 1988 Feb;102(2):369–376. doi: 10.1242/dev.102.2.369. [DOI] [PubMed] [Google Scholar]
  15. Nah H. D., Rodgers B. J., Kulyk W. M., Kream B. E., Kosher R. A., Upholt W. B. In situ hybridization analysis of the expression of the type II collagen gene in the developing chicken limb bud. Coll Relat Res. 1988 Jul;8(4):277–294. doi: 10.1016/s0174-173x(88)80001-3. [DOI] [PubMed] [Google Scholar]
  16. Oberbäumer I., Laurent M., Schwarz U., Sakurai Y., Yamada Y., Vogeli G., Voss T., Siebold B., Glanville R. W., Kühn K. Amino acid sequence of the non-collagenous globular domain (NC1) of the alpha 1(IV) chain of basement membrane collagen as derived from complementary DNA. Eur J Biochem. 1985 Mar 1;147(2):217–224. doi: 10.1111/j.1432-1033.1985.tb08739.x. [DOI] [PubMed] [Google Scholar]
  17. Sandberg M., Autio-Harmainen H., Vuorio E. Localization of the expression of types I, III, and IV collagen, TGF-beta 1 and c-fos genes in developing human calvarial bones. Dev Biol. 1988 Nov;130(1):324–334. doi: 10.1016/0012-1606(88)90438-1. [DOI] [PubMed] [Google Scholar]
  18. Sandberg M., Vuorio E. Localization of types I, II, and III collagen mRNAs in developing human skeletal tissues by in situ hybridization. J Cell Biol. 1987 Apr;104(4):1077–1084. doi: 10.1083/jcb.104.4.1077. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Sariola H., Timpl R., von der Mark K., Mayne R., Fitch J. M., Linsenmayer T. F., Ekblom P. Dual origin of glomerular basement membrane. Dev Biol. 1984 Jan;101(1):86–96. doi: 10.1016/0012-1606(84)90119-2. [DOI] [PubMed] [Google Scholar]
  20. Senior P. V., Critchley D. R., Beck F., Walker R. A., Varley J. M. The localization of laminin mRNA and protein in the postimplantation embryo and placenta of the mouse: an in situ hybridization and immunocytochemical study. Development. 1988 Nov;104(3):431–446. doi: 10.1242/dev.104.3.431. [DOI] [PubMed] [Google Scholar]
  21. Simon-Assmann P., Bouziges F., Arnold C., Haffen K., Kedinger M. Epithelial-mesenchymal interactions in the production of basement membrane components in the gut. Development. 1988 Feb;102(2):339–347. doi: 10.1242/dev.102.2.339. [DOI] [PubMed] [Google Scholar]
  22. Simon-Assmann P., Bouziges F., Vigny M., Kedinger M. Origin and deposition of basement membrane heparan sulfate proteoglycan in the developing intestine. J Cell Biol. 1989 Oct;109(4 Pt 1):1837–1848. doi: 10.1083/jcb.109.4.1837. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Simon-Assmann P., Kedinger M., Haffen K. Immunocytochemical localization of extracellular-matrix proteins in relation to rat intestinal morphogenesis. Differentiation. 1986;32(1):59–66. doi: 10.1111/j.1432-0436.1986.tb00556.x. [DOI] [PubMed] [Google Scholar]
  24. Solursh M., Jensen K. L. The accumulation of basement membrane components during the onset of chondrogenesis and myogenesis in the chick wing bud. Development. 1988 Sep;104(1):41–49. doi: 10.1242/dev.104.1.41. [DOI] [PubMed] [Google Scholar]
  25. Swalla B. J., Upholt W. B., Solursh M. Analysis of type II collagen RNA localization in chick wing buds by in situ hybridization. Dev Biol. 1988 Jan;125(1):51–58. doi: 10.1016/0012-1606(88)90057-7. [DOI] [PubMed] [Google Scholar]
  26. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Timpl R., Dziadek M. Structure, development, and molecular pathology of basement membranes. Int Rev Exp Pathol. 1986;29:1–112. [PubMed] [Google Scholar]

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

RESOURCES