[Calcium oscillations induced by lindane in peritoneal macrophages of mice: control by the maturation stage of the macrophage].
Mouse resident peritoneal macrophages loaded with Fluo-3 were examined for changes in cytosolic calcium concentration ([Ca2+]i) after stimulation with gamma-hexachlorocyclohexane (Lindane or gamma-HCCH). These studies, realized on macrophage populations, or single cells, by digital imaging microscopy, sought to determine the role of calcium influx on cyclical changes according to maturation stages of macrophages. Single cell analysis of [Ca2+]i changes in macrophages, after gamma-HCCH exposure in 600 microM extracellular calcium, demonstrated that: 1) these [Ca2+]i variations were asynchronous oscillations with the same frequency (1.7 min-1), and 2) these [Ca2+]i variations in macrophages were not at the same [Ca2+]i level. This heterogeneity could be correlated to a cell size partition of the macrophage population (10.1 +/- 0.44 and 11.45 +/- 0.43 microns). In the presence of 100 microM calcium, gamma-HCCH induced a calcium influx into the two subpopulations, but the calcium oscillations appeared only in small macrophages. In the largest ones, [Ca2+]i slowly decreased back down to the basal level. The cell size variation could be correlated to a phenotypic heterogeneity, linked to the differenciation stage of the cell. Peroxydase activity showed that small macrophages were in fact exudate macrophages and the largest ones were resident macrophages. Inhibition of the oscillatory patterns by a decrease in the extracellular calcium concentration ([Ca2+]ext) or by lanthanum chloride (LaCl3) addition is indicative of the important role of calcium influx in the triggering of oscillations. The calcium influx was transient and induced inositol phosphate (InsP3) production in macrophages. The maintainance of these calcium oscillations depended on calcium mobilization from intracellular calcium stores by InsP3, since neomycin and 8-(diethylamino) octyl 3,4,5-trimethoxybenzoate (TMB-8) abolished the oscillations. gamma-HCCH induced a transient calcium entry which triggered phospholipase C activation and the associated [Ca2+]i oscillations. However, we showed that differences in cell responses were observed in relationship with the differentiation stage of the mouse peritoneal macrophages, and with the extracellular calcium concentration.
Pinelli
E
E
Laboratoire de Toxicologie et Sécurité Alimentaire, ENSAT, Avenue de l'Agrobiopole Auzeville-Tolosane 31326 Castanet-Tolosan, France.
Pipy
B
B
Teissié
J
J
Gabriel
B
B
D000814:Aniline Compounds; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002121:Calcium Channel Blockers / Q000494:pharmacology; D002454:Cell Differentiation; D048429:Cell Size; D003600:Cytosol / Q000378:metabolism; D005260:Female; D005456:Fluorescent Dyes; D005707:Gallic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology; D001556:Hexachlorocyclohexane / Q000494:pharmacology*; D007295:Inositol Phosphates / Q000378:metabolism; D017737:Macrophages, Peritoneal / Q000187:drug effects* / Q000378:metabolism*; D051379:Mice; D009355:Neomycin / Q000494:pharmacology; D014966:Xanthenes
D000814:Aniline Compounds; D002121:Calcium Channel Blockers; D005456:Fluorescent Dyes; D007295:Inositol Phosphates; D014966:Xanthenes; C059715:Fluo-3; C006014:8-(N,N-diethylamino)octyl-3,4,5-trimethoxybenzoate; D001556:Hexachlorocyclohexane; D005707:Gallic Acid; D009355:Neomycin; D002118:Calcium