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

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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.

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Acknowledgments

This work was supported by grants (IN201003, IN216107, IN201510, IX200610, and IN216213) from Dirección General de Apoyo al Personal Académico, Universidad Nacional Autónoma de México. Laser scanning confocal microscopy expertise was supported by Biólogo Gabriel Ortiz from the Instituto de Fisiología Celular, Universidad Nacional Autónoma de México and Cirujano Dentista María José Gómora Herrera from the Embryology Department, Facultad de Medicina. Laura Valverde-Islas is a PhD student of the Programa de Doctorado en Ciencias Biomédicas, Universidad Nacional Autónoma de México. We thank M.L.I. Rafael Ibarra for his encouragement and guidance.

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  1. Facultad de Medicina, Departamento de Microbiología y Parasitología, Universidad Nacional Autónoma de México, 04510, México, DF, México

    Olivia Reynoso-Ducoing, Laura Valverde-Islas, Cristina Paredes-Salomon, América Pérez-Reyes, Abraham Landa, Lilia Robert & Javier R. Ambrosio

  2. Facultad de Medicina, Departamento de Bioquímica, Universidad Nacional Autónoma de México, 04510, México, DF, México

    Guillermo Mendoza

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  1. Olivia Reynoso-Ducoing
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Correspondence to Javier R. Ambrosio.

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Cysticerci of T. crassiceps ORF strain Parasites were recovered from experimentally infected mice after infection for 3 months, washed in PBS to eliminate any host residue, and observed under a dissection microscope at ×3 magnification. After stimulation with light, the tapeworms showed intense movement of the bladder wall and intense motility. The cysticerci are full of vesicular fluid. The parasites have an elliptical form with two poles. One pole corresponds to the budding region (in the movie at the left) and the other corresponds to the pore (at the right) that is seen in Fig. 1b after the cytoskeleton of the cysticerci was fluorescently stained. (MPG 1,792 kb)

Flame cells of T. crassiceps ORF strain cysticerci Tissues of live cysticerci were observed with a laser scanning confocal microscope (Olympus FV1000, UPLSAPO; ×60 magnification; NA: 1.35) and the movement of the ciliary tufts of several flame cells was observed with the Fluoview FV1000 software. The ciliary tufts are those stained fluorescent green for α-tubulin in Figs. 1f and 2. (MPG 1,290 kb)

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Reynoso-Ducoing, O., Valverde-Islas, L., Paredes-Salomon, C. et al. Analysis of the expression of cytoskeletal proteins of Taenia crassiceps ORF strain cysticerci (Cestoda). Parasitol Res 113, 1955–1969 (2014). https://doi.org/10.1007/s00436-014-3846-4

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