Hypertonic inhibition of exocytosis in neutrophils: central role for osmotic actin skeleton remodeling.
Hypertonicity suppresses neutrophil functions by unknown mechanisms. We investigated whether osmotically induced cytoskeletal changes might be related to the hypertonic inhibition of exocytosis. Hyperosmolarity abrogated the mobilization of all four granule types induced by diverse stimuli, suggesting that it blocks the process of exocytosis itself rather than individual signaling pathways. Concomitantly, osmotic stress provoked a twofold increase in F-actin, induced the formation of a submembranous F-actin ring, and abolished depolymerization that normally follows agonist-induced actin assembly. Several observations suggest a causal relationship between actin polymerization and inhibition of exocytosis: 1) prestimulus actin levels were inversely proportional to the stimulus-induced degranulation, 2) latrunculin B (LB) prevented the osmotic actin response and restored exocytosis, and 3) actin polymerization induced by jasplakinolide inhibited exocytosis under isotonic conditions. The shrinkage-induced tyrosine phosphorylation and the activation of the Na(+)/H(+) exchanger were not affected by LB. Inhibition of osmosensitive kinases failed to prevent the F-actin change, suggesting that the osmotic tyrosine phosphorylation and actin polymerization are independent phenomena. Thus cytoskeletal remodeling appears to be a key component in the neutrophil-suppressive, anti-inflammatory effects of hypertonicity.
Rizoli
S B
SB
Department of Surgery, Toronto General Hospital, and the University of Toronto, Toronto, Ontario, Canada M5G 1L7.
Rotstein
O D
OD
Parodo
J
J
Phillips
M J
MJ
Kapus
A
A
D000199:Actins / Q000502:physiology*; D003594:Cytoplasmic Granules / Q000502:physiology; D003599:Cytoskeleton / Q000502:physiology*; D005089:Exocytosis / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D006982:Hypertonic Solutions / Q000494:pharmacology*; D007477:Ions; D007552:Isotonic Solutions; D009504:Neutrophils / Q000166:cytology / Q000187:drug effects* / Q000502:physiology*; D009997:Osmotic Pressure; D011108:Polymers / Q000378:metabolism; D014443:Tyrosine / Q000378:metabolism
D000199:Actins; D006982:Hypertonic Solutions; D007477:Ions; D007552:Isotonic Solutions; D011108:Polymers; D014443:Tyrosine