Isolation of pure pellicles containing intact basal bodies of Tetrahymena pyriformis. 1985

A Tiedtke

A new procedure for mass isolation of pure pellicles containing intact basal bodies of Tetrahymena pyriformis is reported. The success of the procedure depends on the elimination of the sticky mucocyst contents before fractionation of the cells, which is induced by Alcian Blue 8GS. Under appropriate ionic conditions greater than 95% of the cells are able to form a capsule by simultaneous extrusion of all mature mucocysts. About 50% of these cells are able to escape from their capsules, which are now devoid of mature mucocysts. These cells are separated from the empty capsules and encapsulated cells by passage through layers of gauze of 10 microns pore size. The fractionation of mucocyst-free cells in homogenization buffer yields pure pellicles, which are retained when the homogenate is sieved through steel sieves of 5 microns pore size. Electron-microscopic controls show that the isolated pellicles are not contaminated with subcellular particles. Cells homogenized in the presence of low concentrations of Triton X-100 yield pellicles consisting of the known cell-surface-associated cytoskeletal elements, together with basal bodies. The cilia are detached just above the kinetosomal plate. The basal bodies of isolated pellicles are obviously undamaged, since all the known structures of native basal bodies are preserved. Even the granular matrix, a labile structure in the lumen of the basal body that probably contains RNA, is preserved.

UI MeSH Term Description Entries
D008854 Microscopy, Electron Microscopy using an electron beam, instead of light, to visualize the sample, thereby allowing much greater magnification. The interactions of ELECTRONS with specimens are used to provide information about the fine structure of that specimen. In TRANSMISSION ELECTRON MICROSCOPY the reactions of the electrons that are transmitted through the specimen are imaged. In SCANNING ELECTRON MICROSCOPY an electron beam falls at a non-normal angle on the specimen and the image is derived from the reactions occurring above the plane of the specimen. Electron Microscopy
D002458 Cell Fractionation Techniques to partition various components of the cell into SUBCELLULAR FRACTIONS. Cell Fractionations,Fractionation, Cell,Fractionations, Cell
D002478 Cells, Cultured Cells propagated in vitro in special media conducive to their growth. Cultured cells are used to study developmental, morphologic, metabolic, physiologic, and genetic processes, among others. Cultured Cells,Cell, Cultured,Cultured Cell
D002502 Centrioles Self-replicating, short, fibrous, rod-shaped organelles. Each centriole is a short cylinder containing nine pairs of peripheral microtubules, arranged so as to form the wall of the cylinder. Centriole
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D013769 Tetrahymena pyriformis A species of ciliate protozoa used extensively in genetic research. Tetrahymena pyriformi,pyriformi, Tetrahymena

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