Alkalinizing the intralysosomal pH inhibits degranulation of human neutrophils.
Degranulation of lysosomes is one of the consequences of neutrophil activation. Regulatory mechanisms of lysosomal secretion are thought to be localized largely in the plasma membrane and cytosol, with the lysosome playing a passive role in secretion. Recent evidence indicates that the intralysosomal pH is highly acidic (pH congruent to 5.5) and is maintained by active transport of H+. We investigated whether changes in the intralysosomal pH altered the availability of lysosomes for exocytosis. Intralysosomal pH in intact neutrophils was monitored with the weakly basic fluorescent probe, 9-aminoacridine (9AA). The weak bases, methylamine, chloroquine, clindamycin, propanolol, and ammonium chloride (0.1-50 mM), caused an alkalinization of the intralysosomal pH as determined by reversal of quenching of 9AA fluorescence. Similarly, each of the weak bases, including ammonium chloride, methylamine, chloroquine, ethylamine, propylamine, propanolol, clindamycin, and dansylcadaverine, inhibited neutrophil degranulation in response to the calcium ionophore A23187, phorbol myristate acetate, or the chemotactic peptide, formyl-methionine-leucine-phenylalanine plus cytochalasin B. These studies indicate that an acid intralysosomal pH is important to the neutrophil secretory response and suggest that the lysosome may play an active part in control of degranulation.
D000328:Adult; D000643:Ammonium Chloride / Q000494:pharmacology; D000001:Calcimycin / Q000494:pharmacology; D003594:Cytoplasmic Granules / Q000502:physiology*; D005089:Exocytosis / Q000187:drug effects; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008247:Lysosomes / Q000502:physiology*; D009113:Muramidase / Q000378:metabolism; D009504:Neutrophils / Q000648:ultrastructure*; D013050:Spectrometry, Fluorescence
D000643:Ammonium Chloride; D000001:Calcimycin; D007770:L-Lactate Dehydrogenase; D009113:Muramidase