Analysis of the expression of cytoskeletal proteins of Taenia crassiceps ORF strain cysticerci (Cestoda)

Abstract

The Taenia crassiceps ORF strain is used to generate a murine model of cysticercosis, which is used for diagnosis, evaluation of drugs, and vaccination. This particular strain only exists as cysticerci, is easily maintained under in vivo and in vitro conditions, and offers an excellent model for studying the cytoskeletons of cestodes. In this study, several experimental approaches were used to determine the tissue expression of its cytoskeletal proteins. The techniques used were microscopy (video, confocal, and transmission electron), one-dimensional (1D) and two-dimensional (2D) electrophoresis, immunochemistry, and mass spectrometry. The tissue expression of actin, tubulin, and paramyosin was assessed using microscopy, and their protein isoforms were determined with 1D and 2D electrophoresis and immunochemistry. Nineteen spots were excised from a proteomic gel and identified by liquid chromatography-tandem mass spectrometry and immunochemistry. The proteins identified were classic cytoskeletal proteins, metabolic enzymes, and proteins with diverse biological functions, but mainly involved in detoxification activities. Research suggests that most noncytoskeletal proteins interact with actin or tubulin, and the results of the present study suggest that the proteins identified may be involved in supporting the dynamics and plasticity of the cytoskeleton of T. crassiceps cysticerci. These results contribute to our knowledge of the cellular biology and physiology of cestodes.

In VitroAnalyses Reveal the Effect of Synthetic Cytokinin Forchlorfenuron (FCF) on a Septin-Like Protein of Taeniid Cysticerci

“…In the present study, changes in the fluorescence of polymerized actin or tubulin were observed in cysticerci tissues following FCF treatment (Supplementary Figure 7). These gross observations may indicate that FCF-induced alterations in SEPT4_Tsm disrupt the polymerization of actin and tubulin in T. crassiceps cysticerci, as we have previously shown [20]. Future studies on the interactions between septins and these cytoskeletal proteins should demonstrate the exact nature of this relationship and its importance for the survival of cysticerci.…”

“…Then, cysticerci were fixed with 4% paraformaldehyde and observed under a microscope at 20× magnification (Diaphot; Nikon, Chiyoda-ku, Tokyo, Japan) equipped with an epifluorescence accessory and the appropriate filters. The indirect effects of FCF over actin and α -tubulin of T. crassiceps cysticerci were evaluated in cryosections with phalloidin–rhodamine and commercial DM1A antibody as described in other studies performed by us [20, 22].…”

“…Then, the resultant suspension was desalted and precipitated at -20°C in a solution containing acetone, 10% tricarboxylic acid, and 20 mM dithiothreitol. The suspension was centrifuged at 28,000 × g for 15 min at 4°C and the resultant pellet was recovered and resuspended in cytoskeletal buffer at 4°C as previously reported [20]. Protein concentration was determined with the DC Protein Assay reagent (Bio-Rad, Hercules, CA, USA).…”

“…Thus, changes to the classical morphology of cestodes could derive from alterations to the appearance of subtegumental and flame cells, as well as the complex and intricate protonephridial ducts visible by electron microscopy. As with actin, tubulin, myosin, and paramyosin [17–20], the effect of any compound on the expression of cytoskeletal proteins can be assessed also in the cysticerci of the T. crassiceps ORF strain.…”

Proteome interrogation using gold nanoprobes to identify targets of arctigenin in fish parasites

“…Loss-of function unc-89 mutants display disorganized myosin thick filaments and reduced motility [58]. Besides, myosin-2 is mainly expressed at muscle fibrils in the parasite tegument and the function is to provide motile force through ATP hydrolysis [59]. Therefore, inhibition of myosin-2 or UNC-89 in parasites results in rapid declines in motility, which are in agreement with our previous observations that bath treatment with ARG resulted in rapid declines of G. kobayashii motility [24].…”