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31P and 23Na nuclear magnetic resonance studies of resting and stimulated mast cells
[摘要]

Exocytosis induced by crosslinking the type I receptor for Fce, domains present on rat mucosal mast cells (RBL-2H3-line) requires the influx of Ca2+ ions and is markedly influenced by the concentration of monovalent cations (K+, Na+ and protons) in their medium. We investigated the role of these ions in coupling the immunological stimulus to secretion using NMR spectroscopy to monitor simultanously intraccllular pH, ATP and Na+ concentrations and the secretory response of living adherent mast cells. Using this methodology we observed that: (i) ATP concentration and intracellular pH are highly regulated and no changes could be resolved in them upon stimulation and during exocytosis. (ii) In the absence of potassium ions in the cells' medium, a decrease is observed in the intracellular pH and ATP concentration and an increase in the Na+ concentration. (iii) From the influx of extracellular Na+ following inhibition of the Na+, K+-ATPase by ouabain, we estimated the inward Na+ current of resting cells to 5 × 107 ions/(cell·s). This value does not vary by more than 10% during exocytosis.

[发布日期]  [发布机构] 
[效力级别]  [学科分类] 生物化学/生物物理
[关键词] RBL-2H3 cell;Nuclear magnetic resonance;Exocytosis;Cytosolic pH;Cytosolic [Na+];Cytosolic [ATP];Ag;antigen;BCECF;2';7'-bis(carboxyethyl)-5(6)-carboxyfluorescein;BSA;bovine serum albumin;FcϵRI;Fcϵ-receptor type I;IgE;immunoglobulin E;Pi;inorganic phosphate;PPP;tripolyphosphate;[Na+]i;intracellular sodium concentration;PME;phosphomonoesters [时效性] 
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