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  • 1
    Digitale Medien
    Digitale Medien
    Springer
    Clinical and experimental nephrology 4 (2000), S. 293-299 
    ISSN: 1437-7799
    Schlagwort(e): Key words Calcium receptor ; HEK293 ; NPS R-568 ; Calcimimetics ; Intracellular Ca2+
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Abstract Background. The Ca2+ receptor (CaR) plays a key role in maintaining Ca2+ homeostasis by its presence in the parathyroid gland and kidney. NPS R-568 (referred to as KRN568 in Japan) is a phenylalkylamine compound that activates the CaR. It has been difficult to study Ca2+-sensing mechanisms because of the lack of cell model systems that express reasonable numbers of CaR coupled to downstream effectors and physiological responses. This study was conducted to evaluate the effects of NPS R-568 on the CaR both in vitro and in vivo. Methods. Western blotting analysis of CaR was performed to confirm the existence of CaR in human embryonic kidney 293 (HEK293) cells expressing human CaR (HuCaR-HEK293). Intracellular Ca2+ concentration ([Ca2+]i) and inositol trisphosphate (IP3) content were measured in HuCaR-HEK293 after the addition of NPS R-568 and other agonists. Male Sprague-Dawley rats received NPS R-568 orally, and plasma Ca2+ levels and serum parathyroid hormone (PTH) levels were determined. Results. Western blotting analysis of the crude plasma membrane fraction prepared from HuCaR-HEK293 identified bands immunoreactive with a human CaR-specific antibody. NPS R-568 dose-dependently and stereoselectively increased [Ca2+]i in HuCaR-HEK293, whereas NPS R-568 had no effects in wild-type HEK293 cells. These effects of NPS R-568 were associated with an increase in cytoplasmic IP3 levels and were abolished in the absence of extracellular Ca2+. Single oral administration of NPS R-568 suppressed PTH secretion, followed by decreased plasma Ca2+ levels, in normal rats. Conclusions. These results suggest that NPS R-568 activates CaR and suppresses PTH secretion in vitro and in vivo.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 2
    ISSN: 1435-232X
    Schlagwort(e): Key words Microcell-mediated chromosome transfer ; Chromosome 10p ; Rodent–human hybrid ; Single transferable fragment ; Tumor suppressor gene
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Abstract The introduction of chromosome 10p into human glioblastoma or prostate cancer cells has been demonstrated to suppress their malignant phenotype, suggesting the presence of glioma or prostate tumor suppressor genes on 10p. As a resource for the fine mapping of these genes, a series of human-rodent hybrid cell lines containing single transferable fragments (STFs) of 10p were constructed. Normal chromosome 10 tagged with a neomycin-resistance gene on its short arm was fragmented by gamma-irradiation of 5–10 krad, transferred into mouse L cells or Chinese hamster ovary cells by microcell-mediated chromosome transfer (MMCT), and then selected against G418. Thirty-three independent rodent-human hybrids carrying various-sized STFs were obtained. Polymerase chain reaction (PCR)-based genotyping revealed that these STFs contained the whole, or portions, of a 43-cM region on 10p14-pter and could be defined by 19 sequence-tagged-site (STS) markers. Using this panel of hybrids as donors for further MMCT, genes on the refined fragments could be transferred into other cells. This hybrid panel would therefore be a useful resource for the fine mapping of the genes on 10p14-pter to segments of about 2.4 cM by functional complementation.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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