Localization of acetylated a-tubulin in Tritrichomonas foetus and Trichomonas vaginalis.

Batista, Claudia; Benchimol, Marlene; Cunha-e-Silva, Narcisa L.; Souza, Wanderley de

doi:10.1247/csf.13.445

Cited by 17 publications

(11 citation statements)

References 19 publications

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“…According to these authors, the axostyle would be composed of labile microtubules, which could be stabilized by incubation of the cells in the presence of taxol (Juliano et al 1986a). However, there are several observations which suggest that the axostyle is formed by stable microtubules: (1) incubation of T. foetus in the presence of colchicine or nocodazole does not disturb the axostyle; (2) the axostyle/pelta complex is a very stable microtubular structure as previously observed during the whole division process and also previously using anti-acetylated-tubulin antibodies (Benchimol and DeSouza 1987;Batista et al 1988;Ribeiro et al 2000;Lopes et al 2001); (3) our observations of trichomonads in all phases of the cell cycle always showed the presence of axostyles, even when pseudocysts are formed under stress conditions (Pereira-Neves et al 2003). Our findings conflict with the observations of others because: (1) the electron micrographs presented by these authors did not enable axostyle visualization; (2) there is no mention of the pseudocyst phase, a form that trichomonads assume under drug treatment (Granger et al 2000;PereiraNeves et al 2003); (3) one anti-tubulin antibody was used whereas our group used a panel of a dozen different antibodies, both monoclonal and polyclonal, obtained from different sources (Lopes et al 2001); (4) the fact that, depending on the cell cycle phase, some microtubules, although present, are not detected by immunofluorescence, was overlooked (Boggild et al 2002;Pereira-Neves et al 2003).…”

Section: Discussionsupporting

confidence: 54%

The effect of drugs on cell structure of Tritrichomonas foetus

Costa

Benchimol

2004

Parasitology Research

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The effects of the microtubule affecting drugs taxol, nocodazole and colchicine on the cell cycle and ultrastructure of Tritrichomonas foetus, a protist parasite of cattle, were studied. Alterations in the cytoskeleton, motility and organellar ultrastructure were followed using anti-tubulin antibodies and fluorescence microscopy, scanning- and transmission-electron microscopy. Flow cytometry was also used to analyze the effect of the drugs on the cell cycle. T. foetus was treated with 10 microM taxol, 15 microM nocodazole or 1.5 mM colchicine for 12 h. The first effect observed was pseudocyst formation and alterations in cell motility. The cell cycle was affected and the cells have blocked cytokinesis, but not karyokinesis. The behavior of Golgi, hydrogenosomes and vacuoles was analyzed. The following effects were seen following drug treatments: (1) cell motility was altered and flagella internalized; (2) microtubules of the pelta-axostyle complex were not depolymerized and the axostyle assumed a curved form; (3) hydrogenosomes were of abnormal size and shape; (4) cells became multinucleate; (5) the division process was blocked in cytokinesis; (6) autophagic vacuoles containing a large amount of microtubules were seen; (7) axoneme organization was altered; (8) zoids were formed; (9) signs of cell death, such as membrane blebbing, were observed.

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Section: Discussionsupporting

confidence: 54%

The effect of drugs on cell structure of Tritrichomonas foetus

Costa

Benchimol

2004

Parasitology Research

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“…Pole-tonucleus microtubules attach to the nuclear envelope, possibly constituting a subset of membrane tubulins. Our previous studies were in disagreement with these reports, providing evidence that the pelta-axostyle complex is a stable structure and that the two axostyles are present throughout the mitosis (Batista et al 1988;Ribeiro et al 2000;Silva-Filho and de Souza 1986). Some authors (Viscogliosi and Brugerolle 1994) also stated that the flagella were assembled during mitosis.…”

Section: Introductionmentioning

confidence: 65%

Immunolocalization of tubulin isoforms and post-translational modifications in the protists Tritrichomonas foetus and Trichomonas vaginalis

Lopes

Ribeiro

Benchimol

2001

Histochem Cell Biol

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In the present report we show the distribution of multiple tubulin isoforms in Trichomonas vaginalis and Tritrichomonas foetus, flagellated parasitic protists of the urogenital tracts of human and cattle, respectively, using immunofluorescence and immunoelectron microscopy. We used several monoclonal and polyclonal anti-tubulin antibodies from different sources and recognizing variant tubulin isoforms. Our results demonstrate that: (1) there is a heterogeneous distribution of the different tubulin isoforms in the main microtubular cell structures, such as axostyle, flagella, basal bodies, and mitotic spindle, (2) the axostylepelta junction is a structure with high affinity for glutamylated tubulin antibodies in T. foetus, (3) the spindle labeling is positive to anti-glutamylated tubulin and anti-αtubulin (TAT1 and purchased from Amersham) antibodies in T. vaginalis but it is negative in T. foetus, (4) the nuclear matrix and the cytosol presented positive reaction using glutamylated and TAT1 (anti-α-tubulin) antibodies only in T. vaginalis, and (5) the Golgi complex exhibited staining using the glutamylated tubulin antibody. The present data corroborate with the idea of the existence of a heterogeneous population of microtubules in these protists and of a subset of intracytoplasmic microtubules. Microtubule diversity may reflect distinct tubulins, diverse microtubuleassociated proteins, or a combination of both.

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“…The monoclonal antibody 61 1B1 characterized as reacting specifically with acetylated a-tubulin has been used in a broad spectrum of unicellular and multicellular organisms. The 61 1B 1 epitope is present in metamonads such as Giardia lamblia [Soltys and Gupta, 19941 and trichomonads such as Tritrichomonas foetus and Trichomonas vaginalis [Batista et al, 1988; In view of the distribution of post-translational modifications of tubulin and keeping in mind the limitation of the immunological approach for establishing evolutionary conclusions, it seems that primitive eukaryotic cells may have possessed acetylated and glutamylated but non-tyrosinated and non-glycylated microtubules. Hence they would have had, like many existing protists, a highly ordered and cross-linked microtubular cytoskeleton in which microtubules showed a low level of subunit exchange dynamics.…”

Section: Lmmunolocalization Of Glutamylated Tubulinmentioning

confidence: 99%