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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Methods in cell science 9 (1985), S. 73-81 
    ISSN: 1573-0603
    Keywords: cell dissociation ; clonal culture ; retinal pigmented ; cell differentiation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The original procedure for the dissociation of chick retinal pigmented epithelium (RPE) has been modified in an attempt to increase the yield and plating efficiency of RPE cells and to obtain a more uniform and reproducible pattern of growth and differentiation in clonal culture. The new procedure allows efficient dissociation of very small amounts of. RPE into single cells and the establishment of clonal cultures from the RPE of individual chick embryos or from individual RPE clones. The dissociated cells attach and spread within a few hours after plating and form typical clones with highly pigmented centres within 2 to 3 wk. Individual clones started from the same material show little variation in growth pattern and provide an excellent model system for the study of the expression of cell differentiation in vitro.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Journal of Morphology 157 (1978), S. 99-119 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The process of coelomic pouch formation in Pisaster ochraceus was studied with light microscopy, transmission and scanning electron microscopy and time-lapse cinemicrography as well as with the drug cytochalasin B. As in most asteroids, the paired coelomic pouches of Pisaster ochraceus are formed from outpocketing of the archenteron.Arrays of 50 Å microfilaments are found in the presumptive coelomic pouch cells at the apex of the archenteron as well as in the filopodia of the mesenchyme cells. Both cell types undergo active movements throughout the entire process. Treatment of embryos with cytochalasin B (CCB) during coelomic pouch formation results in the loss of cell movements and the regression of the coelomic pouches; this is accompanied by the loss of microfilament arrays in both cell types. Cell movements and microfilament arrays reappear on removal of CCB and coelomic pouch formation resumes.Our evidence suggests that the microfilaments in the presumptive coelomic pouch cells provide the main force for the outpocketing movement. The major role of the microfilament arrays in the filopodia of the mesenchyme cells associated with the coelomic pouches is to determine the definitive shape and location of the pouches.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Journal of Morphology 203 (1990), S. 361-375 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Six different lectins were used to study the carbohydrate nature of the hyaline layer (HL), the external extracellular matrix of the starfish embryo. Thin sections of embryos fixed in the late gastrula stage were incubated with five fluoresceinated lectins: Con A, WGA, RCA, UEA-I, and SBA. All but UEA-I labelled the HL, suggesting that the following sugars are present: mannose and/or glucose, glcNAc and/or Neu5Ac, galactose, and galNAc. The different lectins produced variable degrees of labelling, with WGA, RCA, and SBA producing more intense labelling than Con A. Binding of lectins by the HL was studied at the ultrastructural level by exposing ultrathin sections to the following lectin-gold conjugates: Con A, WGA, PNA, SBA, and LFA. Lectin binding was observed over the various regions of the HL, recognized by Crawford and Abed (J. Morphol. 176:235-246, '86), i.e., the intervillus layer, the supporting layer and the coarse outer meshwork. Local differences in labelling patterns were observed among the various lectins, with SBA labelling all regions intensely, WGA and PNA labelling the supporting layer predominantly, and Con A labelling the HL only lightly. No labelling was observed with LFA. These lectin-labelling patterns in the HL demonstrate the presence of different glycoconjugates in different regions of the HL, suggesting that the layers differ biochemically. The existence of biochemical differences strengthens the idea that each layer may have different functions in the developing starfish embryo.
    Additional Material: 9 Ill.
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  • 4
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Journal of Morphology 206 (1990), S. 147-161 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: When fixed in the presence of alcian blue, extracellular matrix (ECM) in the embryonic asteroid blastocoel can be visualized by light and electron microscopy as a fibrous meshwork encrusted with alcianophilic material. In early to mid-gastrulae, the ECM is associated with the basal laminae underlying the ectoderm and endoderm. It also forms a fibrous meshwork between them in the posterior part of the blastocoel. In early larvae, when mesenchyme cells arrive at the esophagus to differentiate into smooth muscle, very little ECM is associated with the stomach region. In contrast, a meshwork of long ECM strands radiates from the esophageal basal lamina which connects to a dense ECM web associated with the inner aspect of the dorsal ectoderm. This dorsal web is associated, in turn, with numerous long ECM strands which run parallel to the stomodeum. The strands located between the esophagus and the ectoderm appear when the mouth and coeloms form and may be responsible for a constriction of the ectoderm that forms in this region. During late gastrula one population of mesenchyme cells becomes associated with the esophageal region and differentiates into muscle. Most of the other mesenchyme cells stop migrating through the esophageal web at this time. Less alcianophilic material is associated with the esophageal basal lamina, and the ECM adjacent to the esophagus in the late gastrula and early bipinnaria larvae. The arrangement of the ECM elements suggests that they could be involved in controlling the migration of mesenchyme cells, particularly those destined for the esophagus.
    Additional Material: 19 Ill.
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  • 5
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Journal of Morphology 166 (1980), S. 109-126 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The testicular wall and the process of spermatogenesis in the crinoid, Florometra serratissima, has been studied at the fine structural level. The testicular wall is composed of three layers: a perivisceral layer consisting of nerve processes, muscle fibers, and epithelial cells; a haemal sinus containing haemal fluid, collagen-like fibers, and haemocytes; and a germinal layer consisting of germinal and interstitial cells.The germinal layer is elaborated into numerous folds that project into the lumen of the testis and a branch of the haemal channel extends through the core of each fold. Evidence suggesting that nutrients are carried to the testis and germinal cells via the haemal system is presented. Spermatogonia are concentrated around the base of each fold and spermatocytes line the more distal regions. Spermatids occur at the luminal surface of the germinal layer and spermatozoa fill the testicular lumen. Interstitial cells phagocytize spermatozoa and may also transfer nutrients to spermatids.The nucleus of spermatogonia is large and contains one or two nucleoli. The cytoplasm contains numerous organelles, lipid granules, and a distal and proximal centriole, each with a satellite complex. A striated rootlet extends from the distal centriole. During first meiotic prophase, the distal centriole loses its striated rootlet and produces a flagellum, the proximal centriole loses its satellite complex, the nucleolus disappears, and proacrosomal vesicles are synthesized by the Golgi complex. During spermiogenesis, most of the mitochondria appear to fuse to form a single, large mitochondrion, the nuclear chromatin condenses, and superfluous cytoplasm is lost by autophagocytosis. The formation and definitive positioning of the acrosomal vesicle and periacrosomal material at the apex of the nucleus is described in detail.
    Additional Material: 23 Ill.
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  • 6
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The analysis of proteins by Western blotting is frequently limited by the inability of antibodies to recognize antigenic epitopes in proteins of different species. In the present study, we investigated the influence of mild protease digestion on the reactivity of nitrocellulose-blotted proteins and microscopic sections with antibodies produced against analogous proteins of different species. The proteins were partially purified, electrophoresed and blotted by standard procedures. They were then incubated with either trypsin or pepsin, at concentrations ranging from 0.5 to 80 μg/mL, for different time periods prior to reaction with the first antibody. After incubation with the second antibody, the bands were visualized by chemiluminescence. The following antibody/antigen pairs produced no signal by conventional methods but resulted in the appropriate bands following protease treatment: (i) monoclonal antibody (Mab) to human keratin #18 / rat keratin; (ii) Mab to starfish extracellular matrix/ mouse laminin; (iii) rabbit antiserum to human platelet myosin II/ratfish nonmuscle myosin II. In the latter system, protease treatment also revealed previously hidden epitopes in microscopic sections. Appropriate controls demonstrated that the antibodies retained their specificity after the protease treatment of the preparations. Optimal conditions varied and had to be defined for each protein under study. The results suggest that protease treatment of immunoblots reduces the lack of inter-species reactivity between antibodies and antigens.
    Additional Material: 4 Ill.
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  • 7
    Publication Date: 1999-11-01
    Print ISSN: 0730-2312
    Electronic ISSN: 1097-4644
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Published by Wiley
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  • 8
    Publication Date: 1995-02-01
    Print ISSN: 0012-1606
    Electronic ISSN: 1095-564X
    Topics: Biology
    Published by Elsevier
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