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  • 1
    ISSN: 1573-4919
    Keywords: Menkes gene ; Menkes transcript ; Cu-ATPase ; heavy metal binding domain ; rat brain ATPase ; copper homeostasis ; C6 rat glioma
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Rat Atp7a occupied a single open reading frame (27-4502) which coded for a protein of 1492 residues. Rat Atp7a was 98% and 95% identical to published sequences for the mouse and Chinese hamster, respectively, and 94% homologous to human ATP7A. Compared to ATP7A, the rat transcript coded for an additional alanine (A446) in the heavy metal binding (Hmb) domain and showed a 34 bp gap in the 3′ UTR. Based on published sequence data, hydropathic profiles for rat, mouse, Chinese hamster, and human Cu-ATPases were practically identical with the exception of 8 additional amino acid residues between the 4th and 5th Hmb sites in the human. As deduced from amino acid sequence data, Hmb was predicted to have regions with helical and beta structures. All four species had five of the six metal binding sites centered within hydrophobic regions. The comparative analyses suggested that the Hmb region of the molecule could experience numerous amino acid substitutions with no apparent disruption to the ATPase transport function whereas variations to the ATPase domain would be more critical.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1573-7365
    Keywords: lead toxicity ; brain ; astroglia ; copper ; iron ; neuroglia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Astroglia are implicated in the pathogenesis of lead (Pb) neurotoxicity in two capacities: as a lead sink that sequesters lead and as a target for direct cellular damage. A proposed cellular mechanism of Pb neurotoxicity is the alteration of metal concentrations, particularly the intracellular accumulation of Cu2+. We measured Pb uptake and the effects of Pb acetate on intracellular trace metal concentrations in astroglial cultures prepared from 0-to 4-day-old rat cerebral hemispheres. Mature Sprague Dawley and immature Wistar rat astroglia in culture took up lead from the medium. This finding replicatesin vitro the finding reported by others that astroglia in the brain take up Pb. Intracellular Cu and Fe concentrations (micrograms per 2 x 106 cells) were increased fourfold or more by treatment with 100μMPb for 3 days in the cultures of immature astroglia. Cu levels were also increased twofold or more in mature astroglia treated for 1–3 days with 100μMPb. The significance of this finding is that Cu is a potent inhibitor of Na+,K+ -ATPase, an enzyme by which astroglia are thought to remove K+ from the extracellular fluid in the brain. Thus, this finding supports the hypothesis that elevated [Cu], and perhaps [Fe], is a subcellular mechanism of neurotoxicity.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Metabolic brain disease 4 (1989), S. 187-201 
    ISSN: 1573-7365
    Keywords: astrocytes ; lead ; zinc ; cell culture ; trace metals
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Astroglia take up lead (Pb)in vivo andin vitro. In view of the fact that zinc affects both tissue deposition of Pb and clinical signs of Pb intoxication, the present study was carried out to test the effects of various Zn levels on lead toxicity in astroglia. Primary cultures of astroglia from 1- to 3-day-old neonatal rats were divided into three groups and cultured in Waymouth's 752/1 medium with 0, 1, or 2μM ZnCl2-Each group was further divided into two subgroups which were treated with either 0, 29.9, or 32.5μmol of Pb acetate. Cultures were assayed for viability and metal content after 1 and 3 days of continuous exposure to Pb (designated days 1 and 3) as well as 10, 17, and 24 days after the initiation of a 3-day exposure to Pb. The Trypan blue dye exclusion viability assay showed no significant differences between controls and Pb-treated groups except on day 3, at which time the 0 and 2μM Zn groups treated with Pb had reduced viability.3H-Leucine incorporation into acid-precipitable proteins (cpm/μg protein) was unaffected by Pb or Zn except on days 1 and 17, when cultures given 2μM Zn and no Pb showed increased incorporation. Pb-treated cultures showed a reduction in cell number which was partially offset in a dose-dependent manner by the presence of Zn in the medium but not enough to mask completely the reduction caused by Pb. Pb produced the following effects on intracellular trace metal concentrations: (1) increased intracellular [Pb], (2) increased intracellular [Fe], (3) increased intracellular [Cu], and increased intracellular [Zn]. By day 24, intracellular Cu concentrations were normal, but intracellular [Zn] and [Pb] remained elevated in all Pb-treated subgroups. Furthermore, intracellular Fe levels remained increased in the Pb-treated subgroup cultured with 0μM Zn. Zinc showed a protective effect by (1) reducing intracellular Pb levels and (2) delaying or preventing the Pb-induced increase in intracellular [Fe] and [Zn] but not the increase in intracellular [Cu]. These effects became more pronounced with increasing extracellular Zn concentrations, although intracellular Zn levels did not increase in response to extracellular levels. Increased dietary zinc in rats is known to reduce Pb accumulation in organs. Our results extend this observation to cells in culture and, furthermore, suggest that the Pb-Zn interaction is complex and not simply a substitution of Pb by Zn at the point of absorption through the plasma membrane.
    Type of Medium: Electronic Resource
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