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  • 1
    Electronic Resource
    Electronic Resource
    Copenhagen : Munksgaard International Publishers
    Physiologia plantarum 93 (1995), S. 0 
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: We report an effective system for introducing exogenous DNA into cells of embryonic calli of Oryza sativa L. cv. Japonica. Plant cells were pretreated in hypertonic buffer to draw some of the water from the cells and were then put into medium of less negative osmotic potential containing exogenous DNA and treated immediately with a laser microbeam (λ=355 nm) to puncture holes in the cell wall and membrane. The pretreatment of the cells generated a gradient of osmotic pressure between the inside and outside of the cells, which facilitated the uptake of material into cells through the laser perforations. Bright yellow-green fluorescence could be detected inside cells that had been bathed in a solution containing the fluorescent molecule calcein. β-Glucuronidase (GUS) genes were successfully introduced into rice cells as indicated by gene expression both in post-treated cells and in plantlets derived from kanamycin-resistant calli that had been treated by this method.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 0730-2312
    Keywords: Rous sarcoma virus ; chondrocytes ; matrix calcification ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Endochondral bone formation involves the progression of epiphyseal growth plate chondrocytes through a sequence of developmental stages which include proliferation, differentiation, hypertrophy, and matrix calcification. To study this highly coordinated process, we infected growth plate chondrocytes with Rous sarcoma virus (RSV) and studied the effects of RSV transformation on cell proliferation, differentiation, matrix synthesis, and mineralization. The RSV-transformed chondrocytes exhibited a distinct bipolar, fibroblast-like morphology, while the mock-infected chondrocytes had a typical polygonal morphology. The RSV-transformed chondrocytes actively synthesized extracellular matrix proteins consisting mainly of type I collagen and fibronectin. RSV-transformed cells produced much less type X collagen than was produced by mock-transformed cells. There also was a significant reduction of proteoglycan levels secreted in both the cell-matrix layer and culture media from RSV-transformed chondrocytes. RSV-transformed chondrocytes expressed two- to- threefold more matrix metalloproteinase, while expressing only one-half to one-third of the alkaline phosphatase activity of mock infected cells. Finally, RSV-transformed chondrocytes failed to calcify the extracellular matrix, while mock-transformed cells deposited high levels of calcium and phosphate into their extracellular matrix. These results collectively indicate that RSV transformation disrupts the preprogrammed differentiation pattern of growth plate chondrocytes and inhibit chondrocyte terminal differentiation and mineralization. They also suggest that the expression of extracellular matrix proteins, type II and type X collagens, and the cartilage proteoglycans are important for chondrocyte terminal differentiation and matrix calcification. J. Cell. Biochem. 69:453-462, 1998. © 1998 Wiley-Liss, Inc.
    Additional Material: 8 Ill.
    Type of Medium: Electronic Resource
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