Divalent cation and ATP dependent motility of Toxoplasma gondii tachyzoites after mild treatment with trypsin.
Large percentages of Toxoplasma gondii tachyzoites could be induced to display two types of movement associated with active invasive behavior by exposing them for 1 min to 0.002% trypsin in phosphate-buffered saline (PBS). The motile activity, consisting of clockwise rotation around the posterior end (about 20 revolutions per min) and twirling-gliding over a poly-L-lysine substrate (1.2 +/- 0.2 microns/s standard deviation), was observed and recorded by video-enhanced contrast microscopy. The number of active tachyzoites reached a maximum 1 min after trypsinization; the motile response of the population lasted for about 5 min. Activation was prevented by soybean trypsin-inhibitor, and could not be induced again in previously treated specimens. Electronmicroscopy of trypsinized tachyzoites fixed in the presence of ruthenium-red revealed discrete discontinuities of the plasma membrane, which sealed within 90 min after washing with PBS. Treated tachyzoites were able to invade cultured epithelial cells with a higher relative infectivity than that of untreated parasites. Perfusion of trypsinized tachyzoites with 1 mM of either CaCl2 or MgCl2 and 1 mM ATP increased the number of activated parasites to over 60%; on the other hand, all induced motility was inhibited or blocked by agents that chelate divalent cations. The present preparation, which provided the first serial illustrations of T. gondii movements induced by a defined chemical stimulus, may offer a useful experimental model for the study of motility in this parasite.
Mondragon
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Departamento de Biología Celular, Centro de Investigación y de Estudios Avanzados del IPN, México, D.F. México.
Meza
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Frixione
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D000255:Adenosine Triphosphate / Q000494:pharmacology*; D000818:Animals; D002413:Cations, Divalent / Q000494:pharmacology*; D002465:Cell Movement / Q000187:drug effects; D006801:Humans; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D014122:Toxoplasma / Q000187:drug effects / Q000502:physiology*; D014357:Trypsin / Q000494:pharmacology*
D002413:Cations, Divalent; D000255:Adenosine Triphosphate; D014357:Trypsin