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Biomedical subjects

R Kemler

Publications and source records attributed to R Kemler.

At least 109 records · Page 6Linked to original sources

Cell-adhesion molecule uvomorulin during kidney development.

We studied the expression of a cell adhesion molecule during morphogenesis of the embryonic kidney. The 120-kDa glycoprotein, called uvomorulin, is known to be present on a number of epithelia. During the development of the kidney, a mesenchyme is converted into an epithelium when it is properly induced. The uninduced mesenchyme did not express uvomorulin, as judged by immunofluorescence and immunoblotting using previously characterized antibodies. Uvomorulin does not appear in the mesenchyme as a direct consequence of induction. Rather it becomes detectable approximately 12 hr after completion of induction, at 30-36 hr in vitro when the cells adhere to each other. Distinct differences in uvomorulin expression were seen in the different parts of the nephron. In the mesenchymally derived epithelia (glomeruli, tubules), uvomorulin could be detected only in the tubules, whereas the epithelium of the glomeruli remained negative at all stages of development. Our embryonic studies show that these differences arise very early, as soon as the different parts of the nephron can be distinguished morphologically. It is likely that uvomorulin plays a role in the initial adhesion of the differentiating tubule cells. However, we failed to disrupt histogenesis by applying antibodies to the organ cultures of developing tubules although the antibodies penetrated the tissues well and bound to the differentiating cells.

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Comparison of two cell-adhesion molecules, uvomorulin and cell-CAM 105.

Two cell adhesion molecules, cell-CAM 105 and uvomorulin (UM), were compared by analysing their antigenic structures, their activity in cell aggregation assays and their expression in various tissues. Cell-CAM 105 is a membrane glycoprotein which mediates the intercellular adhesion of reaggregating rat hepatocytes, and UM was first described to be involved in the compaction of preimplantation mouse embryos and embryonal carcinoma cells. UM is not only expressed during embryonic development but also in various adult tissues including liver, epithelia of lung, gut, kidney and uterus. A similar distribution for UM was found in rat tissues on cell types where cell-CAM 105 is known to be present. Our studies show that (i) cell-CAM 105 and UM are distinct and different proteins; (ii) uvomorulin is involved in the compaction of rat preimplantation embryos but Fab anti-UM has no effect on reaggregating rat hepatocytes, where Fab anti-cell CAM is effective; (iii) distribution studies show that UM is expressed on a broader range of epithelial cells while cell-CAM 105 is more restricted to hepatocytes and simple epithelia. In cases where both cell adhesion molecules are expressed on the same cell types they can be localized to different parts of the cell surface.

Adenosine Triphosphatases↗

Cell-adhesion molecule uvomorulin is localized in the intermediate junctions of adult intestinal epithelial cells.

Uvomorulin is a cell-adhesion molecule implicated in the compaction process of mouse preimplantation embryos and the aggregation of embryonal carcinoma cells. A rabbit antiserum against purified uvomorulin also reacts with epithelial cells of various adult tissues. In this study, we investigated the localization of uvomorulin on adult intestinal epithelial cells using electron microscopic analyses. Uvomorulin was shown to exhibit a highly restricted localization in the intermediate junctions of these cells. The results are discussed with respect to a possible adhesive function of uvomorulin on intestinal epithelial cells.

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Molecular properties of ECMA 2 and ECMA 3 antigens defined by monoclonal antibodies against embryonal carcinoma cells.

ECMA 2 and ECMA 3 antigens defined by two monoclonal antibodies are preferentially expressed in early embryonic cells of the mouse. The antigens were isolated from F9 embryonal carcinoma cells by detergent solubilization followed by indirect immunoprecipitation. Both antigens were glycoproteins, which, upon extensive pronase digestion, released the high-molecular-weight glycan (embryoglycan). The immunoprecipitation reactions were inhibited by the glycan, indicating that the two antigens were carried by it. Furthermore, binding of anti-ECMA 2 antibody to the glycan was directly demonstrated by a modified Farr's assay. The antigenic determinant of ECMA 2 antigen was found to involve an alpha-galactosyl residue, since alpha-galactosidase from coffee bean, but not other glycosidases, abolished the antigenic activity. Serological experiments indicated that ECMA 2 antigen is different from other alpha-galactosyl antigens, namely blood group B and P1 antigens and an antigen defined by antibodies in the sera of patients with ovarian germ cell tumors.

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The in vitro development of blastocyst-derived embryonic stem cell lines: formation of visceral yolk sac, blood islands and myocardium.

The in vitro developmental potential of mouse blastocyst-derived embryonic stem cell lines has been investigated. From 3 to 8 days of suspension culture the cells form complex embryoid bodies with endoderm, basal lamina, mesoderm and ectoderm. Many are morphologically similar to embryos of the 6- to 8-day egg-cylinder stage. From 8 to 10 days of culture about half of the embryoid bodies expand into large cystic structures containing alphafoetoprotein and transferrin, thus being analagous to the visceral yolk sac of the postimplantation embryo. Approximately one third of the cystic embryoid bodies develop myocardium and when cultured in the presence of human cord serum, 30% develop blood islands, thereby exhibiting a high level of organized development at a very high frequency. Furthermore, most embryonic stem cell lines observed exhibit similar characteristics. The in vitro developmental potential of embryonic stem cell lines and the consistency with which the cells express this potential are presented as aspects which open up new approaches to the investigation of embryogenesis.

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Rabbit antiserum against a purified surface glycoprotein decompacts mouse preimplantation embryos and reacts with specific adult tissues.

A rabbit antiserum against a purified embryonal carcinoma (EC) cell surface glycoprotein interferes with cell-cell interaction in mouse preimplantation embryos. The 123 kD glycoprotein seems not to be an integral membrane component. The reactivity pattern of the antiserum was studied by immunofluorescence on cryostat sections of post-implantation embryos and of adult tissues. During embryonic development positive reactions were found on all epithelial cells, irrespective of their germ layer origin. Epithelial cells of adult tissues--tongue, uterus, gut, kidney, trachea and liver--react with the antibodies. The results are compared with cell-adhesive molecules previously described on EC cells and preimplantation embryos.

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Some structural and functional aspects of the cell adhesion molecule uvomorulin.

The cell adhesion molecule uvomorulin (UM) was analysed by comparing antisera produced against the whole molecule (gp123) with antisera made against fragments of UM. Of the proteins recognized by different anti-UM antisera (molecular weights of 123, 102, 92 and 84 kDa), the 102 kDa molecule is not derived from gp123. The 102 kDa molecule is not glycosylated and is also different from gp123 by peptide map analysis. However, rabbit antisera raised against the purified 102 kDa protein interfered with the aggregation of embryonal carcinoma (EC) cells. Also, a monoclonal antibody selected to interfere with EC cell aggregation recognized the 102 kDa molecule as well as gp123. Thus, the functional site of cell adhesion seems not to be mediated by sugar residues. Experimental evidence is provided suggesting that UM is not only involved in the compaction of preimplantation embryos but seems to be an ubiquitous cell adhesion molecule regulating epithelial cell adhesion mechanisms.

Animals↗

Reactivity of monoclonal antibodies against intermediate filament proteins during embryonic development.

Monoclonal antibodies (mAbs) against a preparation of intermediate filaments from trophoblastoma cells were studied for their reactivity pattern during embryonic development and on adult tissue cells. Up to day 12 of embryonic development, epithelial cells of the three germ layers reacted with these mAbs. Later during development and in adult tissues, positive reactions could be observed only with epithelial cells derived from mesoderm and endoderm. Because of their tissue distribution, the proteins reacting with these mAbs might belong to the keratin family of intermediate filaments or they might represent a new group of intermediate filaments.

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Monoclonal antibodies against trophectoderm-specific markers during mouse blastocyst formation.

Two-dimensional gel electrophoresis has allowed the detection of proteins characteristic of inner cell mass and trophectoderm in mouse blastocyst. Certain of the proteins characterizing trophectoderm copurify with intermediate filaments from trophectoderm and a trophoblastoma cell line. A monoclonal antibody prepared against proteins of these intermediate filaments labels a filament network in trophectoderm but not in inner cell mass cells.

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Association of the H-Y male antigen with beta2-microglobulin on human lymphoid and differentiated mouse teratocarcinoma cell lines.

The expression of the H-Y antigen has been tested on several human lymphoid lines and mouse teratocarcinoma cell lines during differentiation. The human male lymphoid cell line Raji is a very useful target for studies of the H-Y antigen by lymphocytotoxicity test with rat anti-H-Y sera. With a few exceptions, all cells carrying the Y chromosome were H-Y positive. One of the exceptions is the human Daudi cell line which, besides lacking H-Y antigen, also lacks beta2-microglobulin. We have studied a possible association between the H-Y antigen, beta2-microglobulin, and HLA antigen with redistribution experiments. The results strongly suggest that H-Y antigen is not associated with HLA antigens but with beta2-microglobulin.

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Surface antigen in early differentiation.

Addition of Fab fragments from rabbit antiserum to surface antigen F9 to 2-cell stage mouse embryos in culture does not alter cleavage; however, the addition prevents culture does not alter cleavage; however, the addition prevents the formation of compact morulae and blastocysts. A similar effect is observed when Fab fragments are added to already compact 8-cell stage or even older morulae, but disappears at the beginning of blastocoel formation. This effect is reversible: uncompact 30-cell embryos washed free of Fab become compact in a few hours, produce blastocysts, and upon reimplantation into pseudopregnant mothers can produce mice. Development is not altered by divalent anti-F9 antibodies, by Fab fragments from sera directed against other embryo surface antigens, or by succinyl concanavalin A.

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Embryonal carcinoma antigen and the T/t locus of the mouse.

The presence of the F9 antigen and of four other antigens related to the T/t locus of the mouse was investigated by immunofluorescence on preimplantation embryos. In morulae heterozygous for any of these t haplotypes, both the appropriate t antigen and the F9 antigen are expressed. The F9 antigen segregates among the progeny of crosses producing embryos homozygous for some (tw32 and tw5) but not for other haplotypes. It is concluded that (i) whatever the time of action of a t haplotype, its corresponding antigen is expressed during cleavage and (ii) the F9 antigen is specified by a gene(s) in the region of the T/t locus.

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Absence of a serologically detectable association of murine beta2-microglobulin with the embryonic F9 antigen.

The association of murine beta2-microglobulin to the early embryonic F9 antigen has been investigated by indirect immunofluorescence and by radioimmunoassay. Although some cell lines carry both types of molecules, the beta2-microglobulin was not found expressed on primitive teratocarcinoma cells, where F9 antigen was primarily detected. It is concluded that the low m.w. (12,000 daltons) subunit that was reported to be associated to the F9 antigen is not the adult beta2-microglobulin.

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