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Effects of Li+ transport and intracellular binding on Li+/Mg2+ competition in bovine chromaffin cells.
Fonseca, C P; Montezinho, L P; Nabais, C; Tomé, A R; Freitas, H; Geraldes, C F G C; Castro, M M C A.
Afiliación
  • Fonseca CP; Department of Biochemistry and Centre for Neuroscience of Coimbra, University of Coimbra, P.O. Box 3126, 3001-401 Coimbra, Portugal.
Biochim Biophys Acta ; 1691(2-3): 79-90, 2004 May 03.
Article en En | MEDLINE | ID: mdl-15110989
Li(+) transport, intracellular immobilisation and Li(+)/Mg(2+) competition were studied in Li(+)-loaded bovine chromaffin cells. Li(+) influx rate constants, k(i), obtained by atomic absorption (AA) spectrophotometry, in control (without and with ouabain) and depolarising (without and with nitrendipine) conditions, showed that L-type voltage-sensitive Ca(2+) channels have an important role in Li(+) uptake under depolarising conditions. The Li(+) influx apparent rate constant, k(iapp), determined under control conditions by (7)Li NMR spectroscopy with the cells immobilised and perfused, was much lower than the AA-determined value for the cells in suspension. Loading of cell suspensions with 15 mmol l(-1) LiCl led, within 90 min, to a AA-measured total intracellular Li(+) concentration, [Li(+)](iT)=11.39+/-0.56 mmol (l cells)(-1), very close to the steady state value. The intracellular Li(+) T(1)/T(2) ratio of (7)Li NMR relaxation times of the Li(+)-loaded cells reflected a high degree of Li(+) immobilisation in bovine chromaffin cells, similar to neuroblastoma, but larger than for lymphoblastoma and erythrocyte cells. A 52% increase in the intracellular free Mg(2+) concentration, Delta[Mg(2+)](f)=0.27+/-0.05 mmol (l cells)(-1) was measured for chromaffin cells loaded with the Mg(2+)-specific fluorescent probe furaptra, after 90-min loading with 15 mmol l(-1) LiCl, using fluorescence spectroscopy, indicating significant displacement of Mg(2+) by Li(+) from its intracellular binding sites. Comparison with other cell types showed that the extent of intracellular Li(+)/Mg(2+) competition at the same Li(+) loading level depends on intracellular Li(+) transport and immobilisation in a cell-specific manner, being maximal for neuroblastoma cells.
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Células Cromafines / Litio / Magnesio Límite: Animals Idioma: En Revista: Biochim Biophys Acta Año: 2004 Tipo del documento: Article País de afiliación: Portugal Pais de publicación: Países Bajos
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Células Cromafines / Litio / Magnesio Límite: Animals Idioma: En Revista: Biochim Biophys Acta Año: 2004 Tipo del documento: Article País de afiliación: Portugal Pais de publicación: Países Bajos