Chemotaxis of rat mast cells toward adenine nucleotides.
Rat mucosal mast cells express P2 purinoceptors, occupation of which mobilizes cytosolic Ca2+ and activates a potassium conductance. The primary function of this P2 system in mast cell biology remains unknown. Here, we show that extracellular ADP causes morphological changes in rat bone marrow-cultured mast cells (BMMC) typical of those occurring in cells stimulated by chemotaxins, and that the nucleotides ADP, ATP, and UTP are effective chemoattractants for rat BMMC. ADP was also a chemotaxin for murine J774 monocytes. The nucleotide selectivity and pertussis toxin sensitivity of the rat BMMC migratory response suggest the involvement of P2U receptors. Poorly hydrolyzable derivatives of ADP and ATP were effective chemotaxins, obviating a role for adenosine receptors. Buffering of external Ca2+ at 100 nM or reduction of the electrical gradient driving Ca2+ entry (by elevating external K+) blocked ADP-driven chemotaxis, suggesting a role for Ca2+ influx in this process. Anaphylatoxin C5a was a potent chemotaxin (EC50 approximately 0.5 nM) for J774 monocytes, but it was inactive on rat BMMC in the presence or absence of laminin. Ca2+ removal or elevated [K+] had modest effects on C5a-driven chemotaxis of J774 cells, implicating markedly different requirements for Ca2+ signaling in C5a- vs ADP-mediated chemotaxis. This is supported by the observation that depletion of Ca2+ stores with thapsigargin completely blocked migration induced by ADP but not C5a. These findings suggest that adenine nucleotides liberated from parasite-infested tissue could participate in the recruitment of mast cells by intestinal mucosa.
Department of Zoology and Genetics, Iowa State University, Ames 50011, USA. drmike@iastate.edu
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McCloskey
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Fan
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Luther
D000227:Adenine Nucleotides / Q000494:pharmacology / Q000502:physiology*; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology / Q000187:drug effects; D002118:Calcium / Q000378:metabolism / Q000502:physiology; D002464:Cell Migration Inhibition; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology; D048429:Cell Size / Q000187:drug effects; D002633:Chemotaxis / Q000187:drug effects; D015936:Complement C5a / Q000502:physiology; D005110:Extracellular Space / Q000502:physiology; D019204:GTP-Binding Proteins / Q000502:physiology; D007424:Intracellular Fluid / Q000378:metabolism; D007700:Kinetics; D008407:Mast Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D008564:Membrane Potentials / Q000502:physiology; D051379:Mice; D051381:Rats; D011916:Rats, Inbred F344; D018048:Receptors, Purinergic P2 / Q000502:physiology
D000227:Adenine Nucleotides; D018048:Receptors, Purinergic P2; D015936:Complement C5a; D019204:GTP-Binding Proteins; D002118:Calcium