Premium
Actin cytoskeleton demonstration in Trichomonas vaginalis and in other trichomonads
Author(s) -
Brugerolle Guy,
Bricheux Geneviève,
Coffe Gérard
Publication year - 1996
Publication title -
biology of the cell
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.543
H-Index - 85
eISSN - 1768-322X
pISSN - 0248-4900
DOI - 10.1016/s0248-4900(97)86828-1
Subject(s) - biology , trichomonas vaginalis , actin , immunogold labelling , tritrichomonas foetus , trichomonas , phalloidin , microfilament , microbiology and biotechnology , cytoskeleton , ultrastructure , anatomy , biochemistry , cell , genetics , pregnancy , fetus
Summry— The flagellate form of Trichomonas vaginalis ( Tv ) transforms to amoeboid cells upon adherence to coverslips. They grow and their nuclei divide without undergoing cytokinesis, yielding giant cells and a monolayer of Tv F‐actin was demonstrated in Trichomonas vaginalis by fluorescence microscopy using phalloidin and an anti‐actin mAb which labelled the cytoplasm of both the flagellate and amoeboid forms. Comparative electrophoresis and immunoblotting established that the actin band has the same 42 kDa as muscle actin, but 2‐D electrophoresis resolved the actin band into four spots; the two major spots observed were superimposable with major muscle actin isoforms. Electron microscopy demonstrated an ectoplasmic microfibrillar layer along the adhesion zone of amoeboid Tv adhering to coverslips. Immunogold staining, using anti‐actin monoclonal antibodies demonstrated that this layer was mainly composed of actin microfilaments. A comparative immunoblotting study comprising seven trichomonad species showed that all trichomonads studied expressed actin. The mAb Sigma A‐4700 specific for an epitope on the actin C‐terminal sequence labelled only actin of Trichomonas vaginalis, Tetratrichomonas gallinarum, Trichomitus batrachorum and Hypotrichomonas acosta , but not the actin of Tritrichomonas foetus, Tritrichomonas augusta and Monocercomonas sp. This discrimination between a ‘trichomonas branch’ and a ‘tritrichomonas branch’ is congruent with inferred sequence phylogeny from SSu rRNA and with classical phylogeny of trichomonads.