Intracellular alkalinisation in Vero cells parasitised by Trypanosoma cruzi
| Autor: | Agatangelo Soler-Diaz, Nieves Rodriguez-Cabezas, Jaime Lazuen, Javier Cifuentes, Antonio Osuna, Miguel Angel Ferrer González |
|---|---|
| Rok vydání: | 1998 |
| Předmět: |
Neutral red
Trypanosoma cruzi Intracellular pH Population Fluorescence spectrometry 4 4'-Diisothiocyanostilbene-2 2'-Disulfonic Acid Biology Host-Parasite Interactions Microbiology Amiloride chemistry.chemical_compound Chlorocebus aethiops Image Processing Computer-Assisted Animals Ouabain education Vero Cells education.field_of_study Hydrogen-Ion Concentration Flow Cytometry Molecular biology Quaternary Ammonium Compounds Spectrometry Fluorescence Infectious Diseases Vital stain chemistry Neutral Red Cell culture Vero cell Parasitology Intracellular |
| Zdroj: | International Journal for Parasitology. 28:1841-1851 |
| ISSN: | 0020-7519 |
| DOI: | 10.1016/s0020-7519(98)00148-9 |
| Popis: | We studied the intracellular pH of Vero cells parasitised by Trypanosoma cruzi, using different methods: fluorimetric measurement after labelling the cells with the pH-sensitive intracellular fluorescent dye 2',7',-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein, acetoxymethyl ester; flow cytometry; and image analysis after staining the cells with neutral-red vital stain. The results show that the intracellular pH of the parasitised cells rose in comparison with that of the uninfected control cells. A study of the population of parasitised cells made by flow cytometry allowed us to subdivide the cells from the infected cultures into two populations according to their pH as obtained by fluorimetric measurements. Image analysis showed that the cell cytoplasm was more alkaline in the vicinity of the sites containing parasites. Treatment of the parasitised cells with amiloride, ouabain, or with 4.4'-diisothiocyano-2,2'-stilbene disulphate consistently lowered the pH values of the parasitised cells, but not sufficiently to return to the values of the non-parasitised control cells. When the control cells were subject to similar treatments with the inhibitors, only amiloride acidified the cytoplasm to any extent. The basification undergone by the parasitised cells was independent of the transport systems and may be a consequence of the release of NH4+ by the intracellular amastigotes. |
| Databáze: | OpenAIRE |
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