Monoclonal antibody against SSEA-1 is specific for a subpopulation of astrocytes in mouse cerebellum.
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Animals↗
Biomedical subjects
C Lagenaur
Publications and source records attributed to C Lagenaur.
Immunoselection of oligodendrocytes by magnetic beads. I. Determination of antibody coupling parameters and cell binding conditions.
Oligodendrocytes from early postnatal mouse cerebellum were isolated using polyacrylamide-coated magnetic beads carrying monoclonal antibody to 04 cell surface antigen. Oligodendrocytes were enriched to a purity of 91 +/- 4% starting from a mixed cell population containing approximately 1.5% antigen-positive oligodendrocytes. Viability of 04 antigen-positive oligodendrocytes was approximately 90% as judged by exclusion of trypan blue. Oligodendrocytes were recovered after detachment from the beads with a yield of 19 +/- 6% and after collection by centrifugation onto glass coverslips with yields of approximately 6% of all 04 antigen-positive cells. The final cell yield of oligodendrocytes is approximately 8 x 10(5) cells/gram fresh cerebellar tissue.
Acrylamides↗
Changes in expression of glial antigens M1 and C1 after cerebellar injury.
In response to mechanical injury in the adult mouse cerebellum, Bergmann glia and astrocytes of the granular layer exhibit abnormally increased expression of M1 antigen, while expression of C1 antigen in Bergmann glia is reduced. these reciprocal changes in two different astrocytic antigenic determinants (each recognized by monoclonal antibodies) are easily detected in the immediate area of the wound 4 days after the lesion. Although loss of C1 antigen from Bergmann glia remains localized to the area of the wound, abnormal M1 expression becomes widespread in cerebellar astrocytes, also affecting the contralateral side of the injured cerebellum at its peak 8 to 12 days after injury. These findings suggest that previous observations of abnormal expression of the two antigens in the cerebellum of mutant mice (Sommer, I., and M. Schachner (1981) J. Supramol. Struct. 16: 53-74) might be interpreted with the view that a glial reaction to pathological state might be induced by genetically programmed neuronal cell death and/or abnormal development. We therefore postulate that expression of M1 in astrocytes that normally do not express this antigen and repression of C1 in normally positive astrocytes are indicative of a distinct functional state of astroglia, reactive gliosis.
Animals↗
Monoclonal antibody (M2) to glial and neuronal cell surfaces.
A monoclonal antibody designated M2 arose from the fusion of mouse myeloma cells with splenocytes from a rat immunized with particulate fraction from early postnatal mouse cerebellum. Expression of M2 antigen was examined by indirect immunofluorescence on frozen sections of developing and adult mouse cerebellum and on monolayer cultures of early postnatal mouse cerebellar cells. In adult cerebellum, M2 staining outlines the cell bodies of granule and Purkinje cells. A weaker, more diffuse staining is seen in the molecular layer and white matter. In sections of newborn cerebellum, M2 antigen is weakly detectable surrounding cells of the external granular layer and Purkinje cells. The expression of M2 antigen increases during development in both cell types, reaching adult levels by postnatal day 14. At all stages of postnatal cerebellar development, granule cells that have completed migration to the internal granule layer are more heavily stained by M2 antibodies than are those before and in process of migration. In monolayer cultures, M2 antigen is detected on the cell surface of all GFA protein-positive astrocytes and on more immature oligodendrocytes that express 04 antigen but not 01 antigen. After 3 days in culture, tetanus toxin positive neurons begin to express M2 antigen. The same delayed expression of M2 antigen on neurons is observed in cultures derived from mice ranging in age from postnatal day 0 to 10.
Age Factors↗
Recognition of Bergmann glial and ependymal cells in the mouse nervous system by monoclonal antibody.
A monoclonal antibody designated anti-Cl was obtained from a hybridoma clone isolated from a fusion of NS1 myeloma with spleen cells from BALB/c mice injected with homogenate of white matter from bovine corpus callosum. In the adult mouse neuroectoderm, C1 antigen is detectable by indirect immunohistology in the processes of Bergmann glial cells (also called Golgi epithelial cells) in the cerebellum and of Müller cells in the retina, whereas other astrocytes that express glial fibrillary acidic protein in these brain areas are negative for C1. In addition, C1 antigen is expressed in most, if not all, ependymal cells and in large blood vessels, but not capillaries. In the developing, early postnatal cerebellum, C1 antigen is not confined to Bergmann glial and ependymal cells but is additionally present in astrocytes of presumptive white matter and Purkinje cell layer. In the embryonic neuroectoderm, C1 antigen is already expressed at day 10, the earliest stage tested so far. The antigen is distinguished in radially oriented structures in telencephalon, pons, pituitary anlage, and retina. Ventricular cells are not labeled by C1 antibody at this stage. C1 antigen is not detectable in astrocytes of adult or nearly adult cerebella from the neurological mutant mice staggerer, reeler, and weaver, but is present in ependymal cells and large blood vessels. C1 antigen is expressed not only in the intact animal but also in cultured cerebellar astrocytes and fibroblastlike cells. It is localized intracellularly.
Animals↗
Subclass of astroglia in mouse cerebellum recognized by monoclonal antibody.
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Animals↗
Caulobacter flagellar organelle: synthesis, compartmentation, and assembly.
In Caulobacter crescentus biogenesis of the flagellar organelle occurs during one stage of its complex life cycle. Thus in synchronous cultures it is possible to assay the sequential synthesis and assembly of the flagellum and hook in vivo with a combination of biochemical and radioimmunological techniques. The periodicity of synthesis and the subcellular compartmentation of the basal hook and filament subunits were determined by radioimmune assay procedures. Unassembled 27,000-dalton (27K) flagellin was preferentially located in isolated membrane fractions, whereas the 25K flagellin was distributed between the membrane and cytoplasm. The synthesis of hook began before that of flagellin, although appreciable overlap of the two processes occurred. Initiation of filament assembly coincided with the association of newly synthesized hook and flagellin subunits. Caulobacter flagella are unusual in that they contain two different flagellin subunits. Data are presented which suggest that the ratio of the two flagellin subunits changes along the length of the filament. Only the newly synthesized 25K flagellin subunit is detected in filaments assembled during the swarmer cell stage. By monitoring the appearance of flagellar hooks in the culture medium, the time at which flagella are released was determined.
Bacteria↗
Isolation and characterization of Caulobacter crescentus flagellar hooks.
The basal hook structure of the flagellar organelle Caulobacter crescentus was isolated from release flagella. Hook preparations contained a single major proteins species of 73,000 molecular weight and proteins in smaller amounts that may be minor hook components. Hooks isolated from C. crescents CB13B1a and CB15 were immunologically cross-reactive.
Bacteria↗
Adsorption properties of stage-specific Caulobacter phage phiCbK.
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Adsorption↗
Caulobacter crescentus pili: structure and stage-specific expression.
Pili are functionally expressed during the predivisional and swarmer stages of the Caulobacter crescentus differentiation cycle. They appear on the developing swarmer pole and at the same cellular location as flagella and the phiCbK receptor sites. Pili disappear when the swarmer cell differentiates into a stalked cell; this occurs with the loss of flagella and the disappearance of phage receptor sites. C. crescentus CB13B1a pili have been purified and characterized. Monomeric pilin is a protein with an apparent molecular weight of 8,500 that stains weakly with periodic acid-Schiff reagent. The amino acid composition of purified pilin reveals very low quantities of basic amino acids and a complete absence of methionine. Pilin is synthesized throughout the C. crescentus differentiation cycle. Neither free pili nor pilin monomers are detectable in the growth media, suggesting that loss of piliation in the swarmer- to stalked-cell transition occurs via pilus retraction.
Amino Acids↗
Caulobacter flagellins.
Flagellins from 17 Caulobacter strains were partially purified. Their molecular weights and immunological properties indicated that a two-component flagellin system is common among Caulobacters.
Bacteria↗
Physical characterization of Caulobacter crescentus flagella.
Preparations of intact flagella isolated from Caulobacter crescentus CB13B1a were found to contain two protein species of apparent molecular weights 28,000 and 25,000. Both proteins cross-reacted completely with each other and with purified flagella in Ouchterlony double-immunodiffusion assays. The amino acid compositions of the isolated proteins were similar to one another but precluded any precursor-product relationship. Absence of both the 25,000- and 28,000-molecular-weight proteins from a number of nonmotile mutants and the simultaneous reappearance of these proteins in a motile revertant provide further evidence of the relationship of these two proteins to flagellar structure.
Amino Acids↗
Isolation and characterization of Caulobacter crecentus bacteriophage phi Cd1.
A DNA-containing bacteriophage, phiCd1, was isolated from sewage and shown to infect both stalked and swarmer cells of Caulobacter crescentus strain CB13B1a. phiCd1 is a small, icosohedral bacteriophage, 60 nm in diameter, which possesses a short, noncontractile tail, 10 to 12 nm in length. The bacteriophage particle is composed of at least eight structural proteins. phiCd1 nucleic acid exists as a linear duplex of DNA as judged by: (i) thermal denaturation (Tm), (ii) CsCl density gradient centrifugation, and (iii) chemical analysis of its base composition. The DNA is 61% guanosine plus cytosine, has a buoyant density in CsCl of 1.721 +/- 0.001 g/cm3, and denatures sharply at 78.5 C in 0.1 SSC (standard saline citrate) buffer. The S20, w value for the DNA is 34.3 +/- 0.1S as compared with T7 DNA, indicating a molecular weight of about 29 x 10(6).
Cytosine↗