Organelle-depleted human neutrophil cytoplasts used to study fmet-leu-phe receptor modulation and cell function.
Organelle-depleted human neutrophil cytoplasts were used to study N-formylmethionylleucylphenylalanine (fmet-leu-phe) receptor regulation and adaptation, aggregation, chemotaxis, and chemoattractant-elicited changes in shape, volume, and surface charge. Cytoplasts aggregated in response to the chemoattractant fmet-leu-phe, but the magnitude of the response was less than that in neutrophils. Unlike neutrophils, cytoplasts did not exhibit a second wave of aggregation with the addition of cytochalasin B and also failed to disaggregate. In chemotactic assays, cytoplasts migrated poorly into cellulose nitrate filters, and compared with intact neutrophils required a 100-fold greater concentration of fmet-leu-phe to elicit shape change. In contrast to neutrophils, cytoplasts did not decrease their surface charge in response to fmet-leu-phe and did not exhibit an increase in the binding of fmet-leu-[3H]phe after stimulation with phorbol myristate acetate. However, receptor adaptation and induced changes in membrane potential, as measured by using the membrane probe di-O-C5(3), were similar in cytoplasts and in neutrophils. The data suggest that the presence of functional intracellular organelles is required for normal aggregation and disaggregation, chemotaxis, and shape change induced by fmet-leu-phe, and also peptide receptor upregulation in response to secretagogues. The data show that peptide receptor adaptation occurs in the absence of secretory granules and is independent of receptor upregulation .
D002232:Carbocyanines; D002449:Cell Aggregation / Q000187:drug effects; D002634:Chemotaxis, Leukocyte / Q000187:drug effects; D003594:Cytoplasmic Granules / Q000187:drug effects / Q000502:physiology*; D004586:Electrophoresis; D006801:Humans; D007700:Kinetics; D008564:Membrane Potentials / Q000187:drug effects; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000378:metabolism; D009504:Neutrophils / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D009940:Organoids / Q000502:physiology; D011956:Receptors, Cell Surface / Q000187:drug effects; D044042:Receptors, Formyl Peptide; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D002232:Carbocyanines; D011956:Receptors, Cell Surface; D044042:Receptors, Formyl Peptide; C015843:3,3'-dipropyloxadicarbocyanine; D009240:N-Formylmethionine Leucyl-Phenylalanine; D013755:Tetradecanoylphorbol Acetate