The role of the basement membrane in differential expression of keratin proteins in epithelial cells.
Extracellular matrix is considered to play an important role in determining the phenotype of cells with which it interacts. Here we have investigated the possibility that extracellular matrix is involved in specifying the pattern of keratin expression in epithelial cells. For these studies, we have developed an explant system in which epithelial cells from one type of stratified epithelial tissue, namely conjunctiva, are maintained on an extracellular matrix substrate derived from a different tissue, namely cornea. These ocular tissues are ideal for such analyses since they express distinct sets of keratins. For example, bovine conjunctival epithelium processed for immunofluorescence is not recognized by antibody preparations against keratin K3 or K12. In contrast, K3 and K12 antibodies generate intense staining in bovine corneal epithelium. At the immunochemical level, conjunctival cells in situ appear to possess no K12 and only trace amounts of K3, whereas corneal epithelial cells in situ possess both K3 and K12. When conjunctival cells are maintained on a corneal substrate with an intact basement membrane for 10 days in vitro they begin to express keratin K12 as determined by immunofluorescence. On the other hand, conjunctival cells that are maintained on a corneal substrate lacking a basement membrane fail to show staining with K12 antibodies. Conjunctival cells begin to show intense staining using K3 antibodies within about 10 days of being placed in culture regardless of their substrate. These results indicate that basement membrane can play a positive role in determining cell-specific expression of certain keratins such as K12. However, other keratins such as K3 may be "unmasked" and/or their expression may be upregulated simply by placing conjunctival epithelial cells in culture. We speculate that in conjunctiva K3 expression is influenced by certain negative exogenous factors. We discuss the possible means of regulation of keratin expression in our model system.
Department of Cell, Molecular and Structural Biology, Northwestern University Medical School, Chicago, Illinois.
M A
MA
Kurpakus
E L
EL
Stock
J C
JC
Jones
D000818:Animals; D000906:Antibodies; D001485:Basement Membrane / Q000502:physiology; D002417:Cattle; D003094:Collagen / Q000032:analysis / Q000378:metabolism; D003228:Conjunctiva / Q000166:cytology / Q000502:physiology*; D003315:Cornea / Q000166:cytology / Q000502:physiology*; D004591:Electrophoresis, Polyacrylamide Gel; D004847:Epithelial Cells; D004848:Epithelium / Q000378:metabolism; D005109:Extracellular Matrix / Q000502:physiology*; D005455:Fluorescent Antibody Technique; D007633:Keratins / Q000302:isolation & purification / Q000378:metabolism*; D007797:Laminin / Q000032:analysis / Q000378:metabolism; D008970:Molecular Weight
D000906:Antibodies; D007797:Laminin; D007633:Keratins; D003094:Collagen