Skip to main content
The EMBO Journal logoLink to The EMBO Journal
. 1988 Nov;7(11):3321–3329. doi: 10.1002/j.1460-2075.1988.tb03203.x

Villin expression in the visceral endoderm and in the gut anlage during early mouse embryogenesis.

R Maunoury 1, S Robine 1, E Pringault 1, C Huet 1, J L Guénet 1, J A Gaillard 1, D Louvard 1
PMCID: PMC454827  PMID: 2463156

Abstract

Villin is an evolutionarily well conserved, Ca2+ regulated actin-binding protein, and a major structural component of the brush border of specialized absorptive cells. Using paraffin sections and an affinity purified polyclonal anti-villin antibody, we have investigated the early expression of villin during mouse embryogenesis. Villin is first detectable at the early post-implantation stage in visceral endodermal cells at the periphery of the egg cylinder. In this extra embryonic layer, the expression of villin increases and then persists until full term gestation. In the embryo, villin first appears in gut anlage during the axial rotation. Using the same methodology, villin expression is also demonstrated in differentiating embryoid bodies from a teratocarcinoma. Both in extra embryonic and embryonic extracts, villin expression is confirmed by immunoblot and Northern blot analysis which reveal, respectively, a single polypeptide of 93 kd and an mRNA of 3.4 kb in length, two well defined parameters for adult mouse villin gene expression. The results presented here show that paraffin sections allow very sensitive and highly resolutive detection of antigens in early embryogenesis. They provide a detailed developmental profile of villin expression and demonstrate the usefulness of villin as a marker for epithelial cells involved in absorptive processes.

Full text

PDF
3321

Images in this article

Selected References

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

  1. Alicea H. A., Mooseker M. S. Characterization of villin from the intestinal brush border of the rat, and comparative analysis with avian villin. Cell Motil Cytoskeleton. 1988;9(1):60–72. doi: 10.1002/cm.970090107. [DOI] [PubMed] [Google Scholar]
  2. Andrews G. K. Glucocorticoid receptors in murine visceral yolk sac and liver during development. J Steroid Biochem. 1985 Oct;23(4):437–443. doi: 10.1016/0022-4731(85)90190-6. [DOI] [PubMed] [Google Scholar]
  3. Bretscher A., Osborn M., Wehland J., Weber K. Villin associates with specific microfilamentous structures as seen by immunofluorescence microscopy on tissue sections and cells microinjected with villin. Exp Cell Res. 1981 Sep;135(1):213–219. doi: 10.1016/0014-4827(81)90313-x. [DOI] [PubMed] [Google Scholar]
  4. Bretscher A., Weber K. Villin is a major protein of the microvillus cytoskeleton which binds both G and F actin in a calcium-dependent manner. Cell. 1980 Jul;20(3):839–847. doi: 10.1016/0092-8674(80)90330-x. [DOI] [PubMed] [Google Scholar]
  5. Bretscher A., Weber K. Villin: the major microfilament-associated protein of the intestinal microvillus. Proc Natl Acad Sci U S A. 1979 May;76(5):2321–2325. doi: 10.1073/pnas.76.5.2321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
  7. Carboni J. M., Howe C. L., West A. B., Barwick K. W., Mooseker M. S., Morrow J. S. Characterization of intestinal brush border cytoskeletal proteins of normal and neoplastic human epithelial cells. A comparison with the avian brush border. Am J Pathol. 1987 Dec;129(3):589–600. [PMC free article] [PubMed] [Google Scholar]
  8. Chantret I., Barbat A., Dussaulx E., Brattain M. G., Zweibaum A. Epithelial polarity, villin expression, and enterocytic differentiation of cultured human colon carcinoma cells: a survey of twenty cell lines. Cancer Res. 1988 Apr 1;48(7):1936–1942. [PubMed] [Google Scholar]
  9. 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]
  10. Coudrier E., Kerjaschki D., Louvard D. Cytoskeleton organization and submembranous interactions in intestinal and renal brush borders. Kidney Int. 1988 Sep;34(3):309–320. doi: 10.1038/ki.1988.183. [DOI] [PubMed] [Google Scholar]
  11. Coudrier E., Reggio H., Louvard D. Characterization of an integral membrane glycoprotein associated with the microfilaments of pig intestinal microvilli. EMBO J. 1983;2(3):469–475. doi: 10.1002/j.1460-2075.1983.tb01446.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Coudrier E., Reggio H., Louvard D. Immunolocalization of the 110,000 molecular weight cytoskeletal protein of intestinal microvilli. J Mol Biol. 1981 Oct 15;152(1):49–66. doi: 10.1016/0022-2836(81)90095-4. [DOI] [PubMed] [Google Scholar]
  13. Craig S. W., Lancashire C. L. Comparison of intestinal brush-border 95-Kdalton polypeptide and alpha-actinins. J Cell Biol. 1980 Mar;84(3):655–667. doi: 10.1083/jcb.84.3.655. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Craig S. W., Powell L. D. Regulation of actin polymerization by villin, a 95,000 dalton cytoskeletal component of intestinal brush borders. Cell. 1980 Dec;22(3):739–746. doi: 10.1016/0092-8674(80)90550-4. [DOI] [PubMed] [Google Scholar]
  15. Dudouet B., Robine S., Huet C., Sahuquillo-Merino C., Blair L., Coudrier E., Louvard D. Changes in villin synthesis and subcellular distribution during intestinal differentiation of HT29-18 clones. J Cell Biol. 1987 Jul;105(1):359–369. doi: 10.1083/jcb.105.1.359. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Dziadek M., Adamson E. Localization and synthesis of alphafoetoprotein in post-implantation mouse embryos. J Embryol Exp Morphol. 1978 Feb;43:289–313. [PubMed] [Google Scholar]
  17. Figiel A., Schilt J., Dudouet B., Robine S., Dauça M. Stage-specific polypeptides and villin expression during the intestinal epithelium substitution of the metamorphosing amphibian. Differentiation. 1987;36(2):116–124. doi: 10.1111/j.1432-0436.1987.tb00185.x. [DOI] [PubMed] [Google Scholar]
  18. Gaillard J. A., Maunoury R., Buffe D., Rimbaut C. Le marquage immunologique de l'alpha-foetoprotéine dans l'endoderme proximal et sa signification dans les différenciations extra-embryonnaires des tératocarcinomes de la souris. Ann Inst Pasteur Immunol. 1985 May-Jun;136C(3):353–374. [PubMed] [Google Scholar]
  19. Garrels J. I., Gibson W. Identification and characterization of multiple forms of actin. Cell. 1976 Dec;9(4 Pt 2):793–805. doi: 10.1016/0092-8674(76)90142-2. [DOI] [PubMed] [Google Scholar]
  20. Gerke V., Weber K. Isolation and characterization of mammalian villin and fimbrin, the two bundling proteins of the intestinal microvilli. Eur J Cell Biol. 1983 Sep;31(2):249–255. [PubMed] [Google Scholar]
  21. Glenney J. R., Jr, Bretscher A., Weber K. Calcium control of the intestinal microvillus cytoskeleton: its implications for the regulation of microfilament organizations. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6458–6462. doi: 10.1073/pnas.77.11.6458. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Glenney J. R., Jr, Geisler N., Kaulfus P., Weber K. Demonstration of at least two different actin-binding sites in villin, a calcium-regulated modulator of F-actin organization. J Biol Chem. 1981 Aug 10;256(15):8156–8161. [PubMed] [Google Scholar]
  23. Glenney J. R., Jr, Kaulfus P., Weber K. F actin assembly modulated by villin: Ca++-dependent nucleation and capping of the barbed end. Cell. 1981 May;24(2):471–480. doi: 10.1016/0092-8674(81)90338-x. [DOI] [PubMed] [Google Scholar]
  24. Godbout R., Ingram R. S., Tilghman S. M. Fine-structure mapping of the three mouse alpha-fetoprotein gene enhancers. Mol Cell Biol. 1988 Mar;8(3):1169–1178. doi: 10.1128/mcb.8.3.1169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Godefroy O., Huet C., Blair L. A., Sahuquillo-Merino C., Louvard D. Differentiation of a clone isolated from the HT29 cell line: polarized distribution of histocompatibility antigens (HLA) and of transferrin receptors. Biol Cell. 1988;63(1):41–55. [PubMed] [Google Scholar]
  26. Gröne H. J., Weber K., Helmchen U., Osborn M. Villin--a marker of brush border differentiation and cellular origin in human renal cell carcinoma. Am J Pathol. 1986 Aug;124(2):294–302. [PMC free article] [PubMed] [Google Scholar]
  27. Hammer R. E., Krumlauf R., Camper S. A., Brinster R. L., Tilghman S. M. Diversity of alpha-fetoprotein gene expression in mice is generated by a combination of separate enhancer elements. Science. 1987 Jan 2;235(4784):53–58. doi: 10.1126/science.2432657. [DOI] [PubMed] [Google Scholar]
  28. Hsu S. M., Raine L., Fanger H. Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures. J Histochem Cytochem. 1981 Apr;29(4):577–580. doi: 10.1177/29.4.6166661. [DOI] [PubMed] [Google Scholar]
  29. Huet C., Sahuquillo-Merino C., Coudrier E., Louvard D. Absorptive and mucus-secreting subclones isolated from a multipotent intestinal cell line (HT-29) provide new models for cell polarity and terminal differentiation. J Cell Biol. 1987 Jul;105(1):345–357. doi: 10.1083/jcb.105.1.345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Jackson B. W., Grund C., Schmid E., Bürki K., Franke W. W., Illmensee K. Formation of cytoskeletal elements during mouse embryogenesis. Intermediate filaments of the cytokeratin type and desmosomes in preimplantation embryos. Differentiation. 1980;17(3):161–179. doi: 10.1111/j.1432-0436.1980.tb01093.x. [DOI] [PubMed] [Google Scholar]
  31. Jackson B. W., Grund C., Winter S., Franke W. W., Illmensee K. Formation of cytoskeletal elements during mouse embryogenesis. II. Epithelial differentiation and intermediate-sized filaments in early postimplantation embryos. Differentiation. 1981;20(3):203–216. doi: 10.1111/j.1432-0436.1981.tb01177.x. [DOI] [PubMed] [Google Scholar]
  32. Kellermann O., Buc-Caron M. H., Gaillard J. Immortalization of precursors of endodermal, neuroectodermal and mesodermal lineages, following the introduction of the simian virus (SV40) early region into F9 cells. Differentiation. 1987;35(3):197–205. doi: 10.1111/j.1432-0436.1987.tb00169.x. [DOI] [PubMed] [Google Scholar]
  33. King B. F. The organization of actin filaments in the brush border of yolk sac epithelial cells. J Ultrastruct Res. 1983 Dec;85(3):329–337. doi: 10.1016/s0022-5320(83)90044-8. [DOI] [PubMed] [Google Scholar]
  34. 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]
  35. Lawson K. A., Pedersen R. A. Cell fate, morphogenetic movement and population kinetics of embryonic endoderm at the time of germ layer formation in the mouse. Development. 1987 Nov;101(3):627–652. doi: 10.1242/dev.101.3.627. [DOI] [PubMed] [Google Scholar]
  36. Martin G. R. Teratocarcinomas as a model system for the study of embryogenesis and neoplasia. Cell. 1975 Jul;5(3):229–243. doi: 10.1016/0092-8674(75)90098-7. [DOI] [PubMed] [Google Scholar]
  37. Meehan R. R., Barlow D. P., Hill R. E., Hogan B. L., Hastie N. D. Pattern of serum protein gene expression in mouse visceral yolk sac and foetal liver. EMBO J. 1984 Aug;3(8):1881–1885. doi: 10.1002/j.1460-2075.1984.tb02062.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Melton D. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. doi: 10.1093/nar/12.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Miki A., Kugler P. Comparative enzyme histochemical study on the visceral yolk sac endoderm in the rat in vivo and in vitro. Histochemistry. 1984;81(4):409–415. doi: 10.1007/BF00514337. [DOI] [PubMed] [Google Scholar]
  40. Moll R., Robine S., Dudouet B., Louvard D. Villin: a cytoskeletal protein and a differentiation marker expressed in some human adenocarcinomas. Virchows Arch B Cell Pathol Incl Mol Pathol. 1987;54(3):155–169. doi: 10.1007/BF02899208. [DOI] [PubMed] [Google Scholar]
  41. Mooseker M. S., Graves T. A., Wharton K. A., Falco N., Howe C. L. Regulation of microvillus structure: calcium-dependent solation and cross-linking of actin filaments in the microvilli of intestinal epithelial cells. J Cell Biol. 1980 Dec;87(3 Pt 1):809–822. doi: 10.1083/jcb.87.3.809. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. PIERCE G. B., DIXON F. J., Jr Testicular teratomas. II. Teratocarcinoma as an ascitic tumor. Cancer. 1959 May-Jun;12(3):584–589. doi: 10.1002/1097-0142(195905/06)12:3<584::aid-cncr2820120317>3.0.co;2-f. [DOI] [PubMed] [Google Scholar]
  43. Pineiro A., Olivito A. M., Uriel J. Fixation d'acides gras polyinsaturés pa l'alphafoetoprotéine et la sérum albumine de rat. Comparaison avec l'accumulation de ces acides dans le cerveau au cours du dévoloppement post-natal. C R Seances Acad Sci D. 1979 Nov 26;289(14):1053–1056. [PubMed] [Google Scholar]
  44. Poelmann R. E. The head-process and the formation of the definitive endoderm in the mouse embryo. Anat Embryol (Berl) 1981;162(1):41–49. doi: 10.1007/BF00318093. [DOI] [PubMed] [Google Scholar]
  45. Pringault E., Arpin M., Garcia A., Finidori J., Louvard D. A human villin cDNA clone to investigate the differentiation of intestinal and kidney cells in vivo and in culture. EMBO J. 1986 Dec 1;5(12):3119–3124. doi: 10.1002/j.1460-2075.1986.tb04618.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Robine S., Huet C., Moll R., Sahuquillo-Merino C., Coudrier E., Zweibaum A., Louvard D. Can villin be used to identify malignant and undifferentiated normal digestive epithelial cells? Proc Natl Acad Sci U S A. 1985 Dec;82(24):8488–8492. doi: 10.1073/pnas.82.24.8488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. STEVENS L. C. Embryonic potency of embryoid bodies derived from a transplantable testicular teratoma of the mouse. Dev Biol. 1960 Jun;2:285–297. doi: 10.1016/0012-1606(60)90010-5. [DOI] [PubMed] [Google Scholar]
  48. Shibayama T., Carboni J. M., Mooseker M. S. Assembly of the intestinal brush border: appearance and redistribution of microvillar core proteins in developing chick enterocytes. J Cell Biol. 1987 Jul;105(1):335–344. doi: 10.1083/jcb.105.1.335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Soprano D. R., Wyatt M. L., Dixon J. L., Soprano K. J., Goodman D. S. Retinol-binding protein synthesis and secretion by the rat visceral yolk sac. Effect of retinol status. J Biol Chem. 1988 Feb 25;263(6):2934–2938. [PubMed] [Google Scholar]
  50. West A. B., Isaac C. A., Carboni J. M., Morrow J. S., Mooseker M. S., Barwick K. W. Localization of villin, a cytoskeletal protein specific to microvilli, in human ileum and colon and in colonic neoplasms. Gastroenterology. 1988 Feb;94(2):343–352. doi: 10.1016/0016-5085(88)90421-0. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES