title: Isolation and properties of the axopodial cytoskeleton of a heliozoan, Echinosphaerium akamae.
abstract: The axopodia of a large heliozoan, Echinosphaerium akamae, were efficiently liberated from the cell body by treatment with 65% D(2)O solution containing 5 mM MgCl(2),2.1 mM EGTA, 1 mM KCl and 5 mM HEPES (pH 6.9). After D20 treatment, the cell bodies were removed by centrifugation at a low speed for 30 sec and the resulting supernatant was recentrifuged at 10,000 × g for 10 min. The axopodia were obtained as the pellet fraction without any contamination from the cell body. The isolated axopodia maintained their regular arrangement of cytoskeletal microtubules and were shortened by treatment with Ca(2+). When the isolated axopodia was subjected to SDS-polyacrylamide gel electrophoresis, two major protein bands were detected. The molecular weights of the proteins, tentatively identified as the heliozoan tubulins, were estimated to be about 46 and 50 kD, and an antibody against rat brain tubulin reacted with only the 46 kD protein species.
authors.affiliation: Institute of Pharmaceutical Sciences, Hiroshima University School of Medicine, Hiroshima, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Sugiyama
authors.forename: S
authors.initials: S
authors.lastname: Ikegawa
authors.forename: E
authors.initials: E
authors.lastname: Masuyama
authors.forename: T
authors.initials: T
authors.lastname: Suzaki
authors.forename: M
authors.initials: M
authors.lastname: Ishida
authors.forename: Y
authors.initials: Y
authors.lastname: Shigenaka