Origin and Deposition of Basement Membrane Heparan Sulfate Proteoglycan in the Developing Intestine

• Published in: The Journal of cell biology 1989-10, Vol.109 (4), p.1837-1848
• Main Authors: Simon-Assmann, P., Bouziges, F., Vigny, M., Kedinger, M.
• Format: Article
• Language: English
• Subjects: Animals; Antibodies; Antibodies, Monoclonal; Basement membrane; Basement Membrane - ultrastructure; Biological and medical sciences; Cell coat. Cell surface; Cell structures and functions; Chick Embryo; Chondroitin Sulfate Proteoglycans - analysis; Chondroitin Sulfate Proteoglycans - metabolism; collagen; Embryos; Endoderm - cytology; Epithelial cells; Fetus; Fluorescent Antibody Technique; Fundamental and applied biological sciences. Psychology; Glycosaminoglycans - metabolism; heparan sulfate; Heparan Sulfate Proteoglycans; Heparitin Sulfate - analysis; Heparitin Sulfate - metabolism; Intestines; Intestines - cytology; Intestines - embryology; mesenchyme; Molecular and cellular biology; Molecular Weight; Molecules; Mucosa; Proteoglycans - metabolism; Rats; Rats, Inbred Strains; Spleen cells; Tumors
• ISSN: 0021-9525; 1540-8140
• DOI: 10.1083/jcb.109.4.1837

The deposition of intestinal heparan sulfate proteoglycan (HSPG) at the epithelial-mesenchymal interface and its cellular source have been studied by immunocytochemistry at various developmental stages and in rat/chick interspecies hybrid intestines. Polyclonal heparan sulfate antibodies were produced by immunizing rabbits with HSPG purified from the Engelbreth-Holm-Swarm mouse tumor; these antibodies stained rat intestinal basement membranes. A monoclonal antibody (mAb 4 C1) produced against lens capsule of 11-d-old chick embryo reacted with embryonic or adult chick basement membranes, but did not stain that of rat tissues. Immunoprecipitation experiments indicated that mAb 4 C1 recognized the chicken basement membrane HSPG. Immunofluorescent staining with these antibodies allowed us to demonstrate that distribution of HSPG at the epithelial-mesenchymal interface varied with the stages of intestinal development, suggesting that remodeling of this proteoglycan is essential for regulating cell behavior during morphogenesis. The immunofluorescence pattern obtained with the two species-specific HSPG antibodies in rat/chick epithelial/mesenchymal hybrid intestines developed as grafts (into the coelomic cavity of chick embryos or under the kidney capsule of adult mice) led to the conclusion that HSPG molecules located in the basement membrane of the developing intestine were produced exclusively by the epithelial cells. These data emphasize the notion already gained from previous studies, in which type IV collagen has been shown to be produced by mesenchymal cells (Simon-Assmann, P., F. Bouziges, C. Arnold, K. Haffen, and M. Kedinger. 1988. Development (Camb.). 102:339-347), that epithelial-mesenchymal interactions play an important role in the formation of a complete basement membrane.