Separation of calcareous corpuscles from plerocercoids of Spirometra mansoni and their binding proteins

Abstract

Calcareous corpuscles, known as mineral concretions in Platyhelminthes, may have special biological functions. In the present study we succeeded in the quantitative separation of calcareous corpuscles in plerocercoids of Spirometra mansoni (sparganum) using Ficoll. Purified calcareous corpuscles were reacted with crude extracts of Platyhelminthes, including sparganum. Proteins commonly binding to calcareous corpuscles were molecules of 10, 17, 22, and 35 kDa. A 95-kDa molecule was found to bind specifically in cystic fluid of Cysticercus cellulosae, as was a 40-kDa protein in crude extracts of Taenia saginata and a 27-kDa molecule in crude extracts of Paragonimus westermani and Clonorchis sinensis, respectively. This finding suggests that calcareous corpuscles might have a binding activity to proteins in crude extracts of sparganum.

A new method for concentration of proteins in the calcareous corpuscles separated from the spargana of Spirometra erinacei

“…Coomassie blue staining of gels did not allow visualization of the protein bands due to the low protein concentration, and therefore, the more sensitive silver staining was used. It is known that purified calcareous corpuscles of spargana can bind proteins from other species of Platyhelminthes as shown by an artificial protein binding assay (Yang, 2000). Compared to the results of the experimental protein binding assay, protein molecules extracted from calcareous corpuscles in the present study seemed to bind to the corpuscles passively due to their inorganic components.…”

“…2) differ considerably from those of Yang (2000) in molecular size and abundance. The proteins observed at 35, 22, 17 and 10 kDa were found to bind to corpuscles (Yang, 2000). In the present study, these seem to correspond to 35, 21, 16, and 10 kDa proteins, respectively.…”

“…In particular, the proteins are regarded as important components of the structural framework or to act as a substrate for mineral deposition (Nieland and von Brand, 1969). The presence of corpuscular proteins was first demonstrated in worm sections by histochemical staining (Chowdhury et al, 1955;von Brand et al, 1960), and purified corpuscles showed protein binding activities against crude extracts of Platyhelminthes (Yang, 2000). However, no reports are available on the protein profiles of calcareous corpuscles.…”

“…About 10 volumes of sterile PBS were then added and the remaining white pellet was re-suspended. Calcareous corpuscles were separated using Ficoll-Paque plus solution (Amersham Biosciences, Uppsala, Sweden) (Yang, 2000). Spargana debris was observed under a light microscope.…”

Ultrastructure of the neodermal sclerites of Gyrocotyle urna Grube and Wagener, 1852 (Gyrocotylidea, Cestoda)

“…amphilinids, monozoic and polyzoic tapeworms (Xylander 2001). Calcareous corpuscles are made up of an organic matrix incrusted with inorganic material and may store calcium and magnesium carbonate as well as phosphates and phospholipids (Smyth 1969;Chowdhury and DeRycke 1977;Mackiewicz and Ehrenpsis 1980;McCullough and Fairweather 1987;Yamane et al 1988;Smyth and McManus 1989;Smith and Richards 1993;Yang 2000;Machnicka et al 2005). Whereas the fine morphology of the calcareous corpuscles of both Amphilinidea as well as Eucestoda and that of Gyrocotyle sclerites appears to be quite similar, their location and, most probably, their production are different.…”