Low level expression of cytokeratins 8, 18 and 19 in vascular smooth muscle cells of human umbilical cord and in cultured cells derived therefrom, with an analysis of the chromosomal locus containing the cytokeratin 19 gene.
Of the various intermediate filament (IF) proteins certain cytokeratins, usually a hallmark of epithelial differentiation, can also be detected in some non-epithelial cells in low amounts. We have studied a representative case of this atypical expression, the smooth muscle cells of the blood vessel walls of the human umbilical cord, at the protein and nucleic acid level, by light and electron microscopic immunolocalization, gel electrophoresis and immunoblotting of cytoskeletal proteins, and mRNA identification by Northern blotting. For the latter we have used sensitive probes for various cytokeratins, including new probes for cytokeratin 19. We also describe the chromosome 17 locus comprising the genes for cytokeratins 15 and 19, and we emphasize the occurrence of several unusual and evolutionarily stable sequence elements in the introns of the cytokeratin 19 gene. Most, perhaps all smooth muscle cells of these blood vessels, positively identified by the presence of desmin and smooth muscle type alpha-actin, are immunostained by antibodies specific for cytokeratins 8 and 18, and a subpopulation also contains cytokeratin 19. Immunoelectron microscopy indicates that these cytokeratins are arranged in IFs that are distributed differently from the majority of the IFs formed by desmin and vimentin. Gel electrophoresis of cytoskeletal proteins from microdissected vascular wall tissue shows that the amounts of cytokeratins 8 and 18 present in these tissues are very low, representing less than 1% of the total IF protein, and that cytokeratin 19 is present only in trace amounts. Correspondingly, the contents of mRNAs for cytokeratins 8, 18 and 19 in these tissues are much lower than those present in epithelial cells examined in parallel. We have also established cell cultures derived from umbilical cord vascular smooth muscles that have maintained the expression of cytokeratins 8, 18 and 19, together with vimentin and the smooth muscle type alpha-actin, but do not synthesize desmin. In these cell cultures the cytokeratins are present in much higher amounts than in the original tissue and form IFs that, surprisingly, show a similar distribution as the vimentin IFs and, upon treatment of the cells with colcemid, collapse into juxtanuclear aggregates, often even more effectively than the vimentin IFs do. We conclude that in a certain subtype of smooth muscle cells, the genes encoding cytokeratins of the "simple epithelial type", i.e., cytokeratins 8, 18 and 19, are expressed and that the low level expression of these genes is compatible with myogenic differentiation.(ABSTRACT TRUNCATED AT 400 WORDS)
Bader
B L
BL
Institute of Cell and Tumor Biology, German Cancer Research Center, Heidelberg, Federal Republic of Germany.
Jahn
L
L
Franke
W W
WW
D001483:Base Sequence; D002478:Cells, Cultured; D002874:Chromosome Mapping; D002888:Chromosomes, Human, Pair 19; D003598:Cytoskeletal Proteins / Q000032:analysis; D004586:Electrophoresis / Q000379:methods; D005786:Gene Expression Regulation; D006801:Humans; D007150:Immunohistochemistry; D007633:Keratins / Q000235:genetics* / Q000378:metabolism; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D014470:Umbilical Cord / Q000098:blood supply / Q000166:cytology* / Q000378:metabolism
D003598:Cytoskeletal Proteins; D012333:RNA, Messenger; D007633:Keratins