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1.
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) 相似文献
2.
Cytokeratins in certain endothelial and smooth muscle cells of two taxonomically distant vertebrate species, Xenopus laevis and man 总被引:13,自引:0,他引:13
Lothar Jahn Bernadette Fouquet Klaus Rohe Werner W. Franke 《Differentiation; research in biological diversity》1987,36(3):234-254
Using immunolocalization techniques, electron microscopy, and gel electrophoresis combined with immunoblotting, we have noted remarkable interspecies differences in the expression of cytokeratins in certain nonepithelial cells. In the present study we describe, in two taxonomically distant vertebrate species, the African clawed toad Xenopus laevis and man, endothelial and smooth muscle cells which express cytokeratin intermediate filaments (IFs), in addition to vimentin and/or desmin IFs. In Xenopus, all endothelia seem to produce both vimentin and cytokeratin IFs. As well, certain smooth muscle bundles located in the periphery of the walls of the esophagus and the urinary bladder produce small amounts of cytokeratin IFs in addition to IFs containing vimentin or desmin or both. The amphibian equivalents of human cytokeratins 8 and 18 have been identified in these nonepithelial tissues. In human endothelial cells, immunocytochemical reactions with certain cytokeratin antibodies are restricted to a rare subset of blood vessels. Vessels of this type were first noted in synovial and submucosal tissues, but also occur in some other locations. Cytokeratins have also been detected in certain groups of smooth muscles, such as those present in the walls of some blood vessels in synovial tissue and umbilical cord. Here, the synthesis of low levels of cytokeratins 8 and 18, sometimes with traces of cytokeratin 19, has been demonstrated in smooth muscle cells by colocalization with myogenic marker proteins, such as desmin and/or the smooth-muscle-specific isoform of alpha-actin. Possible reasons for the differences in cytokeratin expression between adjacent endothelia in man, and smooth-muscle structures in both species, as well as biologic and histodiagnostic implications of these findings, are discussed. 相似文献
3.
Cytoskeletal components of lymphoid organs 总被引:24,自引:0,他引:24
Using light and electron microscopic immunolocalization with antibodies to cytoskeletal proteins, we have characterized the nonlymphoid cells of various human lymphoid organs (lymph nodes, tonsils, spleen). In all these tissues, the lymphoid follicles contain a three-dimensional meshwork of "dendritic reticulum cells" which are characterized by the presence of desmosomal junctions, as demonstrated by positive punctate staining with antibodies to the desmosome-specific proteins desmoplakin I and desmoglein, and by intermediate-sized filaments (IFs) of the vimentin type only. In contrast, the extrafollicular regions are characterized by an extended meshwork of other types of reticulum cells, which also contain vimentin IFs but lack desmosomal proteins. In addition, a considerable, although variable proportion of these extrafollicular reticulum cells forms IFs containing cytokeratins 8 and 18 and/or desmin-containing IFs. The occurrence of cytokeratins 8 and 18 in lymph nodes has also been shown by gel electrophoresis and immunoblotting. Results of double-label immunolocalization indicate that some of the extrafollicular reticulum cells coexpress all three kinds of IF protein. A large proportion of these cells also synthesizes another marker of myogenic differentiation, i.e., the isoform of alpha-actin specific for smooth muscle. This proportion includes some cells that are negative for desmin. Comparison of the distribution of cells expressing cytokeratins and/or desmin with that of reticulum cells showing strong alkaline phosphatase activity (as a marker for the so-called "fiber-associated (fibroblastic) reticulum cells") suggests that the former represent a subset of the latter. The biological meaning of these different patterns of expression in reticulum cells and of the resulting cell-type heterogeneity as well as possible implications of these observations for tumor diagnosis, notably of lymph-node metastases and lymphomas, are discussed. 相似文献
4.
Jos L. V. Broers Mirjam Klein Rot G. Peter Vooijs Frans C. S. Ramaekers Lou de Leij Anita ter Haar E. Birgitte Lane Irene M. Leigh Sjoerd Sc. Wagenaar 《Differentiation; research in biological diversity》1989,40(2):119-128
The expression patterns of intermediate filament proteins in fetal and normal or nonpathological adult human lung tissues are described using (chain-specific) monoclonal antibodies. In early stages of development (9-10 weeks and 25 weeks of gestation) only so-called simple cytokeratins such as cytokeratins 7 (minor amounts). 8, 18 and 19 are detected in bronchial epithelial cells. At later stages of development, the cytokeratin expression patterns become more complex. The number of bronchial cells positive for cytokeratin 7 increases, but basal cells in the bronchial epithelium remain negative. These latter cells show, however, expression of cytokeratin 14 in the third trimester of gestation. Developing alveolar epithelial cells express cytokeratins 7, 8, 18 and 19. In adult human bronchial epithelium cytokeratins 4 (varying amounts), 7, 8, 13 (minor amounts), 14, 18 and 19 can be detected, with the main expression of cytokeratins 7, 8, and 18 in columnar cells and the main expression of cytokeratin 14 in basal cells. Vimentin is detected in all mesenchymal tissues. In addition, fetal lung expresses vimentin in bronchial epithelium, however, to a lesser extent with increasing age, resulting in the expression of vimentin in only few scattered bronchial cells at birth. Also in adult bronchial epithelium the expression of vimentin is noticed in part of the basal and columnar epithelial cells. Desmin filaments, present in smooth muscle cells of the lung, appear to alter their protein structure with age. In early stages of development smooth muscle cells surrounding blood vessels are partly reactive with some cytokeratin antibodies and with a polyclonal desmin antibody. At week 9-10 and week 25 of gestation a monoclonal antibody to desmin, however, is not reactive with blood vessel smooth muscle cells but is only reactive with smooth muscle cells surrounding bronchi. With increasing age the reactivity of cytokeratin antibodies with smooth muscle cells in blood vessels decreases, while the reactivity with the monoclonal desmin antibody increases. Our results show that during differentiation profound changes in the intermediate filament expression patterns occur in the different cell types of the developing lung. 相似文献
5.
The expression patterns of individual cytokeratin polypeptides in foetal and adult human pancreatic tissues were examined using monoclonal antibodies. We demonstrated that human pancreatic epithelia in early stages of development (14 weeks of gestation) contain cytokeratins 7, 8, 18 and 19, which are typical of simple epithelia, as well as cytokeratin 4 and 17, which are characteristic of stratified epithelia. In the pancreatic ducts, most of these cytokeratins appeared to be expressed together. Cytokeratins 1, 5, 10, 13, 16 and 20 were not detectable. In contrast, the pancreatic parenchyma was only positive for cytokeratins 8 and 18, except a transient expression of cytokeratins 7 and 19 in pancreatic islets and acinar cells during the foetal development. A focal cytokeratin 7 staining of single acinar cells was seen in newborn and in adult islets. In the stromal tissue, vascular smooth muscle cells were partly reactive with cytokeratin 8 and 18 specific antibodies. The results are discussed in the light of differentiation-dependent changes in the expression of individual cytokeratin polypeptides in developing epithelia. 相似文献
6.
Summary The expression patterns of individual cytokeratin polypeptides in foetal and adult human pancreatic tissues were examined using monoclonal antibodies. We demonstrated that human pancreatic epithelia in early stages of development (14 weeks of gestation) contain cytokeratins 7, 8, 18 and 19, which are typical of simple epithelia, as well as cytokeratin 4 and 17, which are characteristic of stratified epithelia. In the pancreatic ducts, most of these cytokeratins appeared to be expressed together. Cytokeratins 1, 5, 10, 13, 16 and 20 were not detectable. In contrast, the pancreatic parenchyma was only positive for cytokeratins 8 and 18, except a transient expression of cytokeratins 7 and 19 in pancreatic islets and acinar cells during the foetal development. A focal cytokeratin 7 staining of single acinar cells was seen in newborn and in adult islets. In the stromal tissue, vascular smooth muscle cells were partly reactive with cytokeratin 8 and 18 specific antibodies. The results are discussed in the light of differentiation-dependent changes in the expression of individual cytokeratin polypeptides in developing epithelia. 相似文献
7.
Erika Schmid Mary Osborn Elisabeth Rungger-Brndle Giulio Gabbiani Klaus Weber Werner W. Franke 《Experimental cell research》1982,137(2):329-340
The presence of intermediate filament proteins in vascular tissue cells has been examined by immunofluorescence microscopy on frozen sections of the aortic wall of diverse vertebrates (rat, cow, human and chicken) and by gel electrophoresis of cytoskeletal proteins from whole aortic tissue or from stripped tunica media of cow and man. Most cells of the aortic wall in these species contain vimentin filaments, including smoooth muscle cells of the tunica media. In addition, we have observed aortic cells that are positively stained by antibodies to desmin. The presence of desmin in aortic tissue has also been demonstrated by gel electrophoresis for rat, cow and chicken. In aortic tissue some smooth muscle cells contain both types of intermediate filament proteins, vimentin and desmin. Bovine aorta contains, besides cells in which vimentin and desmin seem to co-exist, distinct bundles of smooth muscle cells, located in outer regions of the tunica media, which contain only desmin. The results suggest that (i) intermediate-sized filaments of both kinds, desmin and vimentin, can occur in vascular smooth muscle in situ and (ii) smooth muscle cells of the vascular system are heterogeneous and can be distinguished by their intermediate filament proteins. The finding of different vascular smooth muscle cells is discussed in relation to development and differentiation of the vascular system. 相似文献
8.
Summary We report on the discrimination of vascular smooth muscle cells and myoepithelial cells in primary cultures of human breast
tissue. Breast tissue was disaggregated enzymatically and the resulting organoids seeded in monolayer culture on collagen-coated
plastic in serum-free medium CDM3a. Two main types of organoids were present after enzymatic digestion. One resembled small
blood vessels and the other interlobular ducts or acini of the breast gland epithelium. Within 3 to 8 d after plating the
organoids migrated into typical monolayer islets. These monolayer islets were evaluated using phase contrast microscopy and
further tagged with monoclonal antibodies for immunocytochemical demonstration of Factor VIII-related antigen, muscle iso-forms
of actin, type IV collagen, vimentin, desmin, and keratins. It is concluded that vascular smooth muscle cells resembled myoepithelial
cells by expressing vimentin filaments, depositing type IV collagen, and showing immunoreactivity to muscle iso-forms of actin.
However, whereas vascular smooth muscle cells were associated with endothelial cells and sometimes expressed desmin, myoepithelial
cells appeared together with luminal epithelial cells and expressed cytokeratins.
This work was supported by the Danish Medical Research Council, the Danish Cancer Society, the NOVO Foundation, and the Thaysen
Foundation. 相似文献
9.
Localization of cytokeratins in tissues of the rainbow trout: Fundamental differences in expression pattern between fish and higher vertebrates 总被引:7,自引:0,他引:7
Using a panel of antibodies against different cytokeratins in immunofluorescence microscopy on frozen tissue sections and two-dimensional gel electrophoresis of cytoskeletal proteins from these tissues, we have studied the tissue distribution of cytokeratins in a fish, the rainbow trout Salmo gairdneri. We have distinguished at least 14 different cytokeratin polypeptides in only a limited number of tissues, thus demonstrating the great complexity of the cytokeratin pattern in a fish species. The simplest cytokeratin pattern was that present in hepatocytes, comprising one type-II (L1) and two type-I (L2, L3) polypeptides that appear to be related to mammalian cytokeratins 8 and 18, respectively. Two or all three cytokeratins of this group were also identified in several other epithelial tissues, such as kidney. Epithelia associated with the digestive tract contained, in addition, other major tissue-specific cytokeratins, such as components D1-D3 (stomach, intestine and swim bladder) and B1 and B2 (biliary tract). With the exception of D1, all these polypeptides were also found in a cultured cell line (RTG-2). Epidermal keratinocytes contained D1 and six other major cytokeratins, termed E1-E6. The most complex cytokeratin pattern was that found in the gill epithelium. Surprisingly, antibodies specific for cytokeratins of the L1-L3 group also reacted with certain cell-sheet-forming tissues that are not considered typical epithelia and in higher vertebrates express primarily, if not exclusively, vimentin. Such tissues were (a) endothelia, including the pillar cells of the "gill filaments", (b) scale-associated cells, and (c) the ocular lens epithelium, and also several nonepithelial cell types, such as (d) fibroblasts and other mesenchymal cells, (e) chondrocytes, (f) certain vascular smooth muscle cells, and (g) astroglial cells of the optic nerve. The differences between the patterns of cytokeratin expression in this fish species and those of higher vertebrates are discussed. It is concluded that the diversity of cytokeratins has already been established in lower vertebrates such as fish, but that the tissue-expression pattern of certain cytokeratins has been restricted during vertebrate evolution. We discuss the value of antibodies specific for individual cytokeratin polypeptides as marker molecules indicating cell and tissue differentiation in fish histology, embryology, and pathology. 相似文献
10.
A B Dodge W F Patton M U Yoon H B Hechtman D Shepro 《Comparative biochemistry and physiology. B, Comparative biochemistry》1991,98(4):461-470
1. Using two-dimensional gel electrophoresis and immunoblotting techniques we systematically document the structural diversity of cytoskeletal proteins in tight and leaky cultured microvascular endothelial cells (MEC). Bovine pulmonary and eel rete mirabile MEC primarily express cytokeratins 8 and 19. Cytokeratins 8 and 18 were found to be prominent in rat pulmonary MEC. Bovine retinal MEC contained cytokeratins 8, 18 and 19. Bovine adrenal MEC contain vimentin as their sole intermediate filament protein. 2. Four principal actin isoforms were resolved in micro/macrovascular endothelial cells as well as in vascular smooth muscle cells. Retinal pericytes expressed three principal actin isoforms. 3. These results indicate that MEC are diverse, highly differentiated cells displaying a large repertoire of cytoskeletal protein profiles suited for specific tissue functions. 相似文献
11.
Transient coexpression of desmin and cytokeratins 8 and 18 in developing myocardial cells of some vertebrate species 总被引:15,自引:0,他引:15
Norbert Kuruc Werner W. Franke 《Differentiation; research in biological diversity》1988,38(3):177-193
During myogenesis the intermediate-sized filament (IF) cytoskeleton is characterized by increasing proportions of desmin. While skeletal and smooth muscle formation occurs in free mesenchymal cells containing vimentin-type IFs, myocardial development starts from a polar epithelium containing cytokeratin IFs and desmosomes. Therefore, we have studied the formation of the epicardium and the myocardium in different vertebrate species, combining light and electron microscopic immunolocalization techniques with gel-electrophoretic analyses of cytoskeletal proteins of microdissected myocardial tissue at differing developmental stages. In this report, we describe results obtained from advanced stages of myocardial differentiation. In all species studied the myocardial cell possess IFs abundant in desmin, often together with smaller amounts of vimentin, and the mesothelial layer of the epicardium contains cytokeratin IFs. However, we have observed remarkable interspecies differences with respect to the occurrence of cytokeratins in embryonic myocardial cells. In fetal human myocardium, from week 10 of pregnancy on, but not in juvenile and adult myocardium, and in chicken myocardium of all stages examined (until several days after hatching) specific immunostaining was seen with certain broad-range cytokeratin antibodies as well as with antibodies specific for cytokeratins 18 (in both species) and 8 (showing significant reaction only in human). This cytokeratin immunoreaction, however, did not appear in IFs extending throughout the cytoplasm or at Z-lines, but was localized in punctate arrays representing aggregates of dense material. The aggregates were often enriched at, but not restricted to, the desmosomal plaques of the intercalated discs. These observations were supported by gel-electrophoretic demonstration of small but significant amounts of cytokeratins 18 (in both species) and 8 (detected only in human) in microdissected myocardial tissue. We also observed cytokeratins in smooth muscle cells of some cardiac blood vessels. In contrast, bovine myocardium of advanced fetal age as well as rat and mouse myocardium (from fetal day 12 on) were negative for cytokeratins with all methods, although epicardial cytokeratin IFs were demonstrable. These observations are discussed in relation to myocardial histogenesis and to general problems of cytokeratin gene expression control in epithelial and nonepithelial cells. 相似文献
12.
The cellular composition of the testicular capsule, seminiferous peritubular tissue, the epithelia as well as periductal muscle cell layers of the excurrent ducts was studied, in sexually mature and active Masked Weaver (Ploceus velatus) birds of the passerine family, Ploceidae. Ultrastructure of the contractile cells in the testicular capsule, peritubular and periductal tissues showed that these cells were smooth muscles of typical morphological characteristics. Variability in the immunohistochemical co-expression of microfilaments and intermediate filaments in the different tissues was evident. Actin and desmin proteins were co-expressed immunohistochemically in the testicular capsule and seminiferous peritubular smooth muscle layer. Actin was singly and very weakly expressed in the rete testis epithelium while cytokeratins and desmin were co-expressed in the epithelium of the excurrent ducts. The periductal muscle layer of all ducts of the epididymis, the ductus deferens as well as the seminal glomus, strongly co-expressed actin and desmin. Vimentin was absent in all cells and tissue types studied. There is clear evidence that the tissues of the male gonad and its excurrent ducts in the Masked Weaver, as has been reported for members of the Galloanserae and Ratitae, contain well-formed contractile tissues whose function would include the transportation of luminal through-flow from the testis into, and through, its excurrent ducts. The microtubule helix in the head and of the mid-piece, of elongating spermatids, as well as of the mature spermatozoa in the various excurrent ducts, including some spermatozoa in the seminal glomus, also co-expressed these three proteins. 相似文献
13.
Summary A light and electron microscope immunocytochemical study and Western blotting analysis has been performed on intermediate filaments (vimentin, desmin and cytokeratins) in the testis of the teleost fish Gambusia affinis holbrooki. An immunoreaction to vimentin was observed in the epithelium of the efferent ducts, testicular canal and their surrounding peritubular cells. Positive vimentin immunostaining was also observed in the cells located around seminiferous tubules (boundary cells), Leydig cells, interstitial fibroblasts, chromatophores, and blood vessel endothelial cells. In contrast to mammals, no vimentin immunoreactivity was found in the Sertoli cells. Immunoreactivity to desmin was weak in the epithelial cells of the efferent ducts and testicular canal and intense in the peritubular cells that surrounded these ducts. Desmin immunoreactivity was also observed in the seminiferous tubule boundary cells. The immunoreactivity was weak in the boundary cells that surrounded germ cell cysts containing spermatogonia or spermatocytes and intense in the boundary cells around cysts with elongated or mature spermatids. Immunoreactivity towards cytokeratins was observed only in testicular blood vessels. Cytokeratin immunolabelling was intense in the endothelium and weak in the vascular smooth muscle cells. No cytokeratin immunoreactivity was found in the Sertoli cells, germ cells, interstitial cells or in the efferent duct epithelium. The absence of intermediate filaments in the Sertoli cells, the absence of cytokeratins in the epithelium of the sperm excretory ducts, and the presence of desmin filaments in these epithelial cells are the most important differences with regards to the intermediate filament phenotype in mammalian testes. 相似文献
14.
Anita C. Knapp Franz X. Bosch Michaela Hergt Caecilia Kuhn Stefanie Winter-Simanowski Erika Schmid Sigrid Regauer Jiri Bartek Werner W. Franke 《Differentiation; research in biological diversity》1989,42(2):81-102
Using immunofluorescence microscopy, we observed that in several established cell culture lines derived from different nonepithelial tissues and species, cells spontaneously emerge, usually at low frequencies, which contain cytoplasmic structures decorated by antibodies specific for cytokeratins 8 and 18. This phenomenon was further examined at both the protein (gel electrophoreses of cytoskeletal proteins, followed by immunoblotting) and the RNA (Northern blots, "nuclear run-on" analysis, in situ hybridization) level. Positive cell lines included simian virus (SV40)-transformed human fibroblasts (HF-SV80, WI-38 VA13), human astrocytic glioma cells (U333 CG/343MG), rat (RVF-SMC) and hamster (BHK-21/13) cells derived from vascular smooth muscle and murine sarcoma MS-180 cells. In two cell lines (HF-SV80 and BHK-21/13), the frequency of the cytokeratin-containing cells and of the cytokeratin fibril arrays per cell was drastically increased upon treatment with 5-azacytidine. The structural appearance of the cytokeratins was variable in the different cell lines but could also differ among cells of the same culture: While small granular or comma-shaped structures or bizarrely shaped filament arrays prevailed in WI-38, RVF and normally grown BHK-21 cells, most of the other lines revealed extended normal-looking, fibrillar arrays. In one line (MS-180), the appearance of cytokeratins was associated with a morphological change, as it was only found in a subpopulation of cells that had lost their typical elongated and spindle-shaped phenotype and assumed a rounded ("coccoid") shape. Our results show that the expression of the genes encoding cytokeratins 8 and 18 is not necessarily restricted to programs of epithelial differentiation and that factors stochastically effective appear in cultured cell lines that allow the synthesis of these cytoskeletal components. Mechanisms possibly involved in this spontaneous and selective advent of cytokeratins 8 and 18 and implications for tumor diagnosis are discussed. 相似文献
15.
Mericskay M Parlakian A Porteu A Dandré F Bonnet J Paulin D Li Z 《Developmental biology》2000,226(2):192-208
The desmin gene encodes an intermediate filament protein that is present in skeletal, cardiac, and smooth muscle cells. This study shows that the 4-kb upstream region of the murine desmin promoter directs expression of a lacZ reporter gene throughout the heart from E7.5 and in skeletal muscle and vascular smooth muscle cells from E9. 5. The distal fragment (-4005/-2495) is active in arterial smooth muscle cells but not in venous smooth muscle cells or in the heart in vivo. It contains a CArG/octamer overlapping element (designated CArG4) that can bind the serum response factor (SRF) and an Oct-like factor. The desmin distal fragment can replace a SM22alpha regulatory region (-445/-126) that contains two CArG boxes, to cis-activate a minimal (-125/+65) SM22alpha promoter fragment in arterial smooth muscle cells of transgenic embryos. lacZ expression was abolished when mutations were introduced into the desmin CArG4 element that abolished the binding of SRF and/or Oct-like factor. These data suggest that a new type of combined CArG/octamer element plays a prominent role in the regulation of the desmin gene in arterial smooth muscle cells, and SRF and Oct-like factor could cooperate to drive specific expression in these cells. 相似文献
16.
Specific interaction of the actin-binding domain of dystrophin with intermediate filaments containing keratin 19
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Cytokeratins 8 and 19 concentrate at costameres of striated muscle and copurify with the dystrophin-glycoprotein complex, perhaps through the interaction of the cytokeratins with the actin-binding domain of dystrophin. We overexpressed dystrophin's actin-binding domain (Dys-ABD), K8 and K19, as well as closely related proteins, in COS-7 cells to assess the basis and specificity of their interaction. Dys-ABD alone associated with actin microfilaments. Expressed with K8 and K19, which form filaments, Dys-ABD associated preferentially with the cytokeratins. This interaction was specific, as the homologous ABD of betaI-spectrin failed to interact with K8/K19 filaments, and Dys-ABD did not associate with desmin or K8/K18 filaments. Studies in COS-7 cells and in vitro showed that Dys-ABD binds directly and specifically to K19. Expressed in muscle fibers in vivo, K19 accumulated in the myoplasm in structures that contained dystrophin and spectrin and disrupted the organization of the sarcolemma. K8 incorporated into sarcomeres, with no effect on the sarcolemma. Our results show that dystrophin interacts through its ABD with K19 specifically and are consistent with the idea that cytokeratins associate with dystrophin at the sarcolemma of striated muscle. 相似文献
17.
Intermediate filaments in smooth muscle 总被引:1,自引:0,他引:1
Tang DD 《American journal of physiology. Cell physiology》2008,294(4):C869-C878
The intermediate filament (IF) network is one of the three cytoskeletal systems in smooth muscle. The type III IF proteins vimentin and desmin are major constituents of the network in smooth muscle cells and tissues. Lack of vimentin or desmin impairs contractile ability of various smooth muscle preparations, implying their important role for smooth muscle force development. The IF framework has long been viewed as a fixed cytostructure that solely provides mechanical integrity for the cell. However, recent studies suggest that the IF cytoskeleton is dynamic in mammalian cells in response to various external stimulation. In this review, the structure and biological properties of IF proteins in smooth muscle are summarized. The role of IF proteins in the modulation of smooth muscle force development and redistribution/translocation of signaling partners (such as p130 Crk-associated substrate, CAS) is depicted. This review also summarizes our latest understanding on how the IF network may be regulated in smooth muscle. cytoskeleton; force development; vimentin; desmin 相似文献
18.
Genes involved in differentiation of notochord or muscle are expressed in specific regions of the involuted dorsal mesoderm in mid-gastrula Xenopus embryo. The presumptive notochord or the presomitic mesoderm have been cultured either in isolation or recombination to investigate whether these tissues have been determined. Cell differentiation was checked by specific markers of notochord (Shh) or muscle cell (desmin, myosin). The results show that the presumptive notochord can differentiate into vacuolated notochord with a weak expression of Shh, while the presomitic mesoderm differentiate into muscle cells with a normal expression of desmin and myosin in vitro. The same result was obtained when the two tissues have been cocultured. These data suggest that the cell fate of the involuted dorsal mesoderm in mid-gastrula has been determined, cells can differentiate according to their fates without further signals from the adjacent tissues, but no functional structures can be formed by these tissues in vitro. 相似文献
19.
Monoclonal antibodies to desmin, the muscle-specific intermediate filament protein 总被引:23,自引:2,他引:21
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A set of monoclonal antibodies to desmin has been isolated from a fusion of mouse myeloma cells with spleen cells from mice immunized with purified porcine desmin. Eleven group I antibodies recognized desmin in the immune blot, and using defined desmin fragments the epitope has been tentatively assigned as lying between residues 325 and 372. When cell lines were tested in immunofluorescence only the human line RD and hamster BHK-21 were positive. When tissue sections were used, skeletal, cardiac, visceral and some vascular smooth muscle cells were positive. Thus, the group I antibodies appear specific for desmin and do not recognize other intermediate filament proteins. Group II monoclonals recognized not only desmin in the immune blot but also other polypeptides. The epitope of this class is located between residues 70 and 280. In immunofluorescence on cell lines and tissues, the staining patterns of group II antibodies were more complicated and demonstrate that not only other intermediate filament proteins but also additional antigenic determinants are being recognized. The group I antibodies stain, as expected from their desmin specificity, rat and human rhabdomyosarcomas and thus appear to be useful reagents in pathology. 相似文献
20.
Expression of cytokeratins 7, 8, 17, 18 in human embryos and fetuses of 6.5-13 weeks was studied using light and electron immunocytochemistry and immunoelectroblotting with the monoclonal antibodies. Cytokeratins 8 and 18 were expressed in 6.5-8 week old embryos not only in epithelium but also in mesenchyme of allantois, urogenital sinus, Wolffian and Mullerian ducts, mesentery, urinary bladder and certain regions of colon, rectum and atrium cordis walls. Furthermore, starting from the 10th week smooth-muscle cells of ring layer in caudal part of rectum bound antibodies against cytokeratin 17 in addition to those against cytokeratins 8 and 18. Corresponding mesenchymal and smooth-muscle cells of adult individuals did not react with either of them. Cytokeratins were still synthesized when mesenchymal cells of embryonic intestine wall were cultivated in vitro. Intermediate filaments of these cells contain cytokeratins 8 and 18, as demonstrated by electron immunocytochemistry and immunoelectroblotting. Thus, the expression of cytokeratins is not restricted to adult and embryonic epithelial tissues but is also characteristic of mesenchyme and smooth muscle differentiation in human embryos and fetuses. 相似文献