After mating, spermatophores of signal crayfish are stored on the body of the female for a period before fertilization. This study compared the post-mating protein profile and pattern of protein tyrosine phosphorylation of the signal crayfish spermatophore to that of the freshly ejaculated spermatophore and found substantial differences. Two major bands of tyrosine-phosphorylated proteins of molecular weights 10 and 50kDa were observed in the freshly ejaculated spermatophore of the signal crayfish. While the tyrosine-phosphorylated protein band with molecular weight 10kDa was formed by protein(s) of similar pH, the band with molecular weight of 50kDa consisted of proteins of varying pH. In the post-mating spermatophore, the band with molecular weight of 50kDa was not detected, and an increase in the level of protein tyrosine phosphorylation was observed in the 10kDa band. The microtubular radial arms of the spermatozoon showed a positive reaction to an anti-tyrosine antibody conjugated with gold particles in both the freshly ejaculated and post-mating spermatophores. In conclusion, the male gamete of the signal crayfish undergoes molecular modification during post-mating storage on the body of the female including changes in the level of protein expression and protein tyrosine phosphorylation. Structural similarity of the radial arms in the crayfish immotile spermatozoon with flagellum, which is the main site of protein tyrosine phosphorylation in the mammalian motile spermatozoa, raises questions regarding evolution and function of such organelles across the animal kingdom that must be addressed in the future studies.

• MeSH
• fosforylace fyziologie   MeSH
• kopulace MeSH
• motilita spermií fyziologie   MeSH
• proteiny genetika   metabolismus   MeSH
• severní raci fyziologie   MeSH
• spermatogonie fyziologie   MeSH
• spermie fyziologie   MeSH
• tyrosin metabolismus   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

The microtubular cytoskeleton of most single-celled eukaryotes radiates from an organizing center called the flagellar apparatus, which is essential for locomotion, feeding and reproduction. The structure of the flagellar apparatus tends to be conserved within diverse clades of eukaryotes, and modifications of this overall structure distinguish different clades from each other. Understanding the unity and diversity of the flagellar apparatus provides important insights into the evolutionary history of the eukaryotic cell. Diversification of the flagellar apparatus is particularly apparent during the multiple independent transitions to parasitic lifestyles from free-living ancestors. However, our understanding of these evolutionary transitions is hampered by the lack of detailed comparisons of the microtubular root systems in different lineages of parasitic microbial eukaryotes and those of their closest free-living relatives. Here we help to establish this comparative context by examining the unity and diversity of the flagellar apparatus in six major clades containing both free-living lineages and endobiotic (parasitic and symbiotic) microbial eukaryotes: stramenopiles (e.g., Phytophthora), fornicates (e.g., Giardia), parabasalids (e.g., Trichomonas), preaxostylids (e.g., Monocercomonoides), kinetoplastids (e.g., Trypanosoma), and apicomplexans (e.g., Plasmodium). These comparisons enabled us to address some broader patterns associated with the evolution of parasitism, including a general trend toward a more streamlined flagellar apparatus.

• MeSH
• biologická evoluce * MeSH
• cytoskelet metabolismus   MeSH
• flagella metabolismus   ultrastruktura   MeSH
• lokomoce MeSH
• mikrotubuly metabolismus   MeSH
• molekulární evoluce MeSH
• paraziti klasifikace   fyziologie   MeSH
• rozmnožování MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

OBJECTIVES: Flubendazole (FLU), a member of benzimidazole family of anthelmintic drugs, is able to inhibit proliferation of various cancer cells. The aim of present study was to elucidate the mechanisms of antiproliferative effect of FLU on colorectal cancer cells in vitro. METHODS: The effect of FLU on proliferation, microtubular network, DNA content, caspase activation and senescence induction was studied in SW480 and SW620 cell lines. KEY FINDINGS: Flubendazole significantly affected cell proliferation in a pattern typical for mitotic inhibitor. This was accompanied by decrease in cyclin D1 levels, increase in cyclin B1 levels, activation of caspase 2 and caspase 3/7 and PARP cleavage. Morphological observations revealed disruption of microtubular network, irregular mitotic spindles, formation of giant multinucleated cells and increase in nuclear area and DNA content. In SW620 cell line, 37.5% giant multinucleated cells induced by FLU treatment showed positivity for SA-β-galactosidase staining. Cell lines were able to recover from the treatment and this process was faster in SW480 cells. CONCLUSION: Flubendazole in low concentration temporarily inhibits cell proliferation and induces mitotic catastrophe and premature senescence in human colon cancer cells in vitro.

• MeSH
• aparát dělícího vřeténka účinky léků   ultrastruktura   MeSH
• lidé MeSH
• mebendazol analogy a deriváty   farmakologie   MeSH
• mikrotubuly účinky léků   ultrastruktura   MeSH
• mitóza účinky léků   MeSH
• nádorové buněčné linie MeSH
• obrovské buňky účinky léků   ultrastruktura   MeSH
• proliferace buněk účinky léků   MeSH
• stárnutí buněk účinky léků   MeSH
• tubulin metabolismus   MeSH
• velikost buněčného jádra účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Many of the highly organized microtubular arrangements in ciliates are located in the cortical area containing membrane vesicles and vacuoles. In Tetrahymena thermophila and Paramecium caudatum, immunofluorescence microscopy with the monoclonal antibody TU-06, directed against beta-tubulin, revealed distinct staining of this cortical region alone, while the cilia and other microtubular structures were unstained. The specificity of the antibody was confirmed by immunoblotting and by preabsorption of the antibody with purified tubulin. Double-label immunofluorescence with antibodies against gamma-tubulin, detyrosinated alpha-tubulin, and centrin showed that the TU-06 epitope is localized outside the basal body region. This was also confirmed by immunogold electron microscopy of thin sections. Proteolytic digestion of porcine brain beta-tubulin combined with a peptide scan of immobilized, overlapping peptides disclosed that the epitope was in the beta-tubulin region beta81-95, a region which is phylogenetically highly conserved. As known posttranslational modifications of beta-tubulin are located outside this area, the observed staining pattern cannot be interpreted as evidence of subcellular sequestration of modified tubulin. The limited distribution of the epitope could rather reflect the dependence of TU-06 epitope exposition on conformations of tubulin molecules in microtubule arrangements or on differential masking by interacting proteins.

• MeSH
• buněčná membrána imunologie   MeSH
• buňky 3T3 MeSH
• epitopy analýza   imunologie   metabolismus   MeSH
• financování organizované MeSH
• imunoblotting MeSH
• mapování epitopu MeSH
• myši MeSH
• Paramecium imunologie   MeSH
• Tetrahymena thermophila imunologie   MeSH
• tubulin imunologie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH

• MeSH
• entomologie MeSH
• fyziologie buňky MeSH
• hematologie MeSH
• kompartmentace buňky MeSH
• nádorové procesy MeSH
• Publikační typ
• kongresy MeSH

• Konspekt
• Biologické vědy
• NLK Obory
• biologie
• biologie
• onkologie