title: Toxoplasma gondii: dithiol-induced Ca2+ flux causes egress of parasites from the parasitophorous vacuole.
abstract: Ca2+ is an essential activator of motility in the obligate intracellular parasite Toxoplasma gondii. Ca2+ ionophore A23187 and intracellular microinjection of Ca2+ initiate motility of parasites residing in parasitophorous vacuoles (PV). The source of Ca2+ and the mechanism by which it activates motility in vivo remain uncertain. Exposure of the parasites to dithiothreitol (DTT) can activate egress of previously nonmotile intravacuolar parasites within 60 sec. DTT is also known to activate both isoforms of the highly concentrated nucleoside triphosphate hydrolase (NTPase) produced by T. gondii. Using an adherent cell analysis system (ACAS) for Ca2+ imaging, a brief 15-50% increase in intra-PV fluorescence ratio was observed after exposure of infected fibroblasts to 5 mM DTT. Chelation of intracellular Ca2+ with BAPTA-AM and extracellular Ca2+ with EGTA blocked the DTT effect; however, this chelation did not prevent the activation of parasites nor the Ca2+ response to the Ca2+ ionophore ionomycin, suggesting that the Ca2+ that activates motility may reside near or within the parasite itself. This result demonstrates that an increase in Ca2+ within the vacuole precedes the onset of motility and the correlation of the DTT effect on motility and tachyzoite NTPase suggests that NTPase activation may be involved in the Ca2+ flux.
authors.affiliation: Department of Medicine (Section of Neurology), Dartmouth Medical School, Lebanon, New Hampshire 03756, USA.
authors.forename: E W
authors.initials: EW
authors.lastname: Stommel
authors.forename: K H
authors.initials: KH
authors.lastname: Ely
authors.forename: J D
authors.initials: JD
authors.lastname: Schwartzman
authors.forename: L H
authors.initials: LH
authors.lastname: Kasper
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002478:Cells, Cultured; D002614:Chelating Agents / Q000378:metabolism / Q000494:pharmacology; D004229:Dithiothreitol / Q000494:pharmacology*; D004347:Drug Interactions; D004533:Egtazic Acid / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology; D005347:Fibroblasts / Q000469:parasitology* / Q000648:ultrastructure; D005456:Fluorescent Dyes / Q000378:metabolism; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007211:Indoles / Q000378:metabolism; D015759:Ionomycin / Q000494:pharmacology; D007476:Ionophores / Q000494:pharmacology; D018613:Microscopy, Confocal; D015203:Reproducibility of Results; D012433:Ryanodine / Q000494:pharmacology; D013439:Sulfhydryl Reagents / Q000494:pharmacology*; D019284:Thapsigargin / Q000494:pharmacology; D014122:Toxoplasma / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D014617:Vacuoles / Q000469:parasitology*
chemical_list: D002614:Chelating Agents; D005456:Fluorescent Dyes; D007211:Indoles; D007476:Ionophores; D013439:Sulfhydryl Reagents; C070379:1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester; D012433:Ryanodine; D004533:Egtazic Acid; D015759:Ionomycin; D019284:Thapsigargin; C048960:indo-1; D002118:Calcium; D004229:Dithiothreitol