Structural Analysis of Basal Bodies of The Isolated Oral Apparatus of Tetrahymena Pyriformis

1970 ◽  
Vol 6 (3) ◽  
pp. 679-700
Author(s):  
J. WOLFE
Keyword(s):  
Structural Analysis ◽  
Cell Membrane ◽  
Basal Body ◽  
Structural Integrity ◽  
Tetrahymena Pyriformis ◽  
Basal Plate ◽  
Basal Bodies ◽  
The Third ◽  
Ionic Detergent

The oral apparatus of Tetrahymena pyriformis was isolated using a non-ionic detergent to disrupt the cell membrane. The mouth consists largely of basal bodies and microfilaments. Each basal body is attached to the mouth by a basal plate which is integrated into the meshwork of microfilaments that confers upon the oral apparatus its structural integrity. Each basal body is composed of 9 triplet microtubules. Two of the 3 tubules, subfibres ‘A’ and ‘B’ are composed of filamentous rows of globules with a spacing of 4.5nm. The third tubule, subfibre ‘C’, is only one-third the length of the basal body.

Development of macrociliary cells in Beroe. I. Actin bundles and centriole migration

1988 ◽  
Vol 89 (1) ◽  
pp. 67-80
Author(s):  
S. Tamm ◽  
S.L. Tamm
Keyword(s):  
Cell Membrane ◽  
Basal Body ◽  
Actin Filaments ◽  
Close Association ◽  
Directed Assembly ◽  
Double Layers ◽  
Basal Bodies ◽  
Apical Surface ◽  
Actin Bundle ◽  
Microfilament Bundle

Differentiation of macrociliary cells on regenerating lips of the ctenophore Beroe was studied by transmission electron microscopy. In this study of early development, we found that basal bodies for macrocilia arise by an acentriolar pathway near the nucleus and Golgi apparatus, in close association with plaques of dense fibrogranular bodies. Procentrioles are often aligned side-by-side in double layers with the cartwheel ends facing outward toward the surrounding plaques of dense granules. Newly formed basal bodies then disband from groups and develop a long striated rootlet at one end. At the same time, an array of microfilaments arises in the basal cytoplasm. The microfilaments are arranged in parallel strands oriented toward the cell surface. The basal body-rootlet units are transported to the apical surface in close association with the assembling actin filament bundle. Microfilaments run parallel to and alongside the striated rootlets, to which they often appear attached. Basal body-rootlet units migrate at the heads of trails of microfilaments, as if they are pushed upwards by elongation of their attached actin filaments. Near the apical surface the actin bundle curves and runs below the cell membrane. Newly arrived basal body-rootlets tilt upwards out of the microfilament bundle to contact the cell membrane and initiate ciliogenesis. The basal bodies tilt parallel to the flat sides of the rootlets, and away from the direction in which the basal feet point. The actin bundle continues to enlarge during ciliogenesis. These results suggest that basal body migration may be driven by the directed assembly of attached actin filaments.

scholarly journals THE NUCLEIC ACIDS OF BASAL BODIES ISOLATED FROM TETRAHYMENA PYRIFORMIS

1965 ◽  
Vol 25 (2) ◽  
pp. 217-228 ◽  
Author(s):  
Eugene J. Hoffman
Keyword(s):  
Electron Microscope ◽  
Nucleic Acids ◽  
Basal Body ◽  
Tetrahymena Pyriformis ◽  
Isolation Procedure ◽  
Basal Bodies ◽  
Phenol Extraction ◽  
Body Self

Pellicular fragments were isolated from ethanol-fixed cells of the holotrichous ciliate Tetrahymena pyriformis by the action of digitonin. The isolated pellicles were further fragmented and the basal bodies of the cilia isolated from them by three methods. The preparations, examined in the electron microscope as embedded sections or negatively stained samples, consisted mainly of somewhat deformed pellicular material, the bulk of which was basal body. DNA was determined by the diphenylamine method and by reaction with DNase, and RNA, by the orcinol method. Nucleic acids were isolated by phenol extraction and analyzed spectrophotometrically and by reaction with RNase. The assays indicated 1.2 to 2.6 per cent RNA, similar to previously published work, but only 0.0 to 1.0 per cent DNA, near enough the sensitivity limits to render the presence of DNA in the preparations uncertain. Although the isolation procedure removed nuclear contents and ribosomes, the nucleic acids could still be a residual contaminant bound to the pellicle during the isolation. Hypotheses of basal body self-duplication, moreover, can be constructed both with and without nucleic acids.

Evidence for the presence of DNA at basal body sites in Tetrahymena pyriformis

1965 ◽  
Vol 162 (989) ◽  
pp. 473-491 ◽  
Keyword(s):  
Cell Division ◽  
Basal Body ◽  
Mitotic Cycle ◽  
Tritiated Thymidine ◽  
Tetrahymena Pyriformis ◽  
Cell Cortex ◽  
Basal Bodies ◽  
Temperature Shock ◽  
Large Numbers ◽  
Order Of Magnitude

The reasons that have led to a search for DNA in the basal body of Tetrahymena pyriformis are twofold: the well-known property of proliferation of this organelle and the possibility that basal body DNA might be involved in its morphogenesis. After a brief review of earlier work the methods employed in this paper are described. To ensure large numbers of cells in a particular state of development organisms were grown in synchronized culture. Animals required for autoradiographic studies were appropriately treated with tritiated thymidine. All investigations were made on the cell cortex or 'ghost’ in order to avoid confusion from cell contents. In addition to autoradiography of ghosts, tests were made with acridine orange in the fluorescence microscope. It is concluded from fluorescence tests that basal bodies of T. pyriformis strain S contain DNA . This DNA is not detectable for the first 2h of the temperature-shock cycle, but is detect­able thereafter until cell division. The presence of DNA is confirmed by the autoradiography experiments. The amount of DNA per basal body is estimated very roughly in order of magnitude as 2 × 10 -16 g. The origin of basal body DNA is discussed and the possibilities and consequences of the existence of DNA in the homologous centriole are examined in terms of the mitotic cycle, the amoeba-flagellate transformation in Naegleria , and artificial parthenogenesis. The paper concludes with a brief discussion of the genetic implications of basal body DNA .

scholarly journals Bld10/Cep135 stabilizes basal bodies to resist cilia-generated forces

2012 ◽  
Vol 23 (24) ◽  
pp. 4820-4832 ◽  
Author(s):  
Brian A. Bayless ◽  
Thomas H. Giddings ◽  
Mark Winey ◽  
Chad G. Pearson
Keyword(s):  
Basal Body ◽  
Structural Integrity ◽  
Basal Bodies ◽  
Ciliary Beating ◽  
Full Complement ◽  
Domain Protein ◽  
Motile Cilia ◽  
The Stability

Basal bodies nucleate, anchor, and organize cilia. As the anchor for motile cilia, basal bodies must be resistant to the forces directed toward the cell as a consequence of ciliary beating. The molecules and generalized mechanisms that contribute to the maintenance of basal bodies remain to be discovered. Bld10/Cep135 is a basal body outer cartwheel domain protein that has established roles in the assembly of nascent basal bodies. We find that Bld10 protein first incorporates stably at basal bodies early during new assembly. Bld10 protein continues to accumulate at basal bodies after assembly, and we hypothesize that the full complement of Bld10 is required to stabilize basal bodies. We identify a novel mechanism for Bld10/Cep135 in basal body maintenance so that basal bodies can withstand the forces produced by motile cilia. Bld10 stabilizes basal bodies by promoting the stability of the A- and C-tubules of the basal body triplet microtubules and by properly positioning the triplet microtubule blades. The forces generated by ciliary beating promote basal body disassembly in bld10Δ cells. Thus Bld10/Cep135 acts to maintain the structural integrity of basal bodies against the forces of ciliary beating in addition to its separable role in basal body assembly.

scholarly journals DEVELOPMENT OF THE FLAGELLAR APPARATUS OF NAEGLERIA

1966 ◽  
Vol 31 (1) ◽  
pp. 43-54 ◽  
Author(s):  
Allan D. Dingle ◽  
Chandler Fulton
Keyword(s):  
Cell Membrane ◽  
Cell Surface ◽  
Basal Body ◽  
Flagellar Apparatus ◽  
Basal Bodies ◽  
Naegleria Gruberi

Flagellates of Naegleria gruberi have an interconnected flagellar apparatus consisting of nucleus, rhizoplast and accessory filaments, basal bodies, and flagella. The structures of these components have been found to be similar to those in other flagellates. The development of methods for obtaining the relatively synchronous transformation of populations of Naegleria amebae into flagellates has permitted a study of the development of the flagellar apparatus. No indications of rhizoplast, basal body, or flagellum structures could be detected in amebae. A basal body appears and assumes a position at the cell surface with its filaments perpendicular to the cell membrane. Axoneme filaments extend from the basal body filaments into a progressive evagination of the cell membrane which becomes the flagellum sheath. Continued elongation of the axoneme filaments leads to differentiation of a fully formed flagellum with a typical "9 + 2" organization, within 10 min after the appearance of basal bodies.

scholarly journals THE MORPHOGENESIS OF BASAL BODIES AND ACCESSORY STRUCTURES OF THE CORTEX OF THE CILIATED PROTOZOAN TETRAHYMENA PYRIFORMIS

1969 ◽  
Vol 40 (3) ◽  
pp. 716-733 ◽  
Author(s):  
Richard D. Allen
Keyword(s):  
Basal Body ◽  
Tetrahymena Pyriformis ◽  
Basal Bodies ◽  
Proximal Surface ◽  
Body Development ◽  
Transmission Electron ◽  
Short Cylinder ◽  
Dividing Cells ◽  
Constant Distance ◽  
Single Tubules

Dividing cells of Tetrahymena pyriformis were observed by transmission electron microscopy for signs of morphogenesis of cortical structures. The earliest stage of basal body development observed was of a short cylinder of nine single tubules connected by an internal cartwheel structure. This is set perpendicular to the mature basal body at its anterior proximal surface under the transverse microtubules and next to the basal microtubules. Sequential stages show that the single tubules become triplet tubules and that the "probasal bodies" then elongate and tilt toward the organism's surface while maintaining a constant distance of 75–100 mµ with the "parent." The new basal body after it is fully extended contacts the pellicle, and then assumes a parallel orientation with and moves anterior to the parent basal body. The electron-opaque core in the lumen of the basal body and accessory structures around its outer proximal surface appear after the developing basal body has elongated. These accessory structures associating with their counterparts from other basal bodies and with the longitudinal microtubules may play a role in the final positioning of basal bodies and thus in the maintenance of cortical patterns. Observations on a second sequence of basal body formation suggest that the oral anlage arises by multiple duplication of somatic basal bodies.

scholarly journals PARTIAL PURIFICATION AND PHOSPHOTUNGSTATE SOLUBILIZATION OF BASAL BODIES AND KINETODESMAL FIBERS FROM TETRAHYMENA PYRIFORMIS

1973 ◽  
Vol 57 (3) ◽  
pp. 601-612 ◽  
Author(s):  
Robert W. Rubin ◽  
William P. Cunningham
Keyword(s):  
Basal Body ◽  
Gel Electrophoresis ◽  
Sucrose Gradient ◽  
Gradient Centrifugation ◽  
Sodium Dodecyl ◽  
Tetrahymena Pyriformis ◽  
Partial Purification ◽  
Basal Bodies ◽  
Thin Sectioning ◽  
Triton X

Previously devised methods for the isolation of basal bodies from ciliate protozoans were found to be inadequate for chemical analysis. We have modified and expanded these procedures and developed a method which gives preparations containing mainly basal bodies and kinetodesmal fibers. This procedure involved fixation of cells in 30% ETOH followed by digitonin or Triton X-100 solubilization and homogenization with a Brinkmann Polytron. This is followed by sucrose gradient centrifugation. Negative staining and thin sectioning revealed these preparations to be substantially more pure than those of previous workers. It was also found that neutralized phosphotungstate (PTA) solubilized many of the components present in fixed Tetrahymena. Neutralized 1.0% PTA solubilized axonemes, cortical, axonemal, and basal body microtubules as well as kinetodesmal fibers. These results have been confirmed by both electron microscope observations and gel electrophoresis of 100,000 g supernatants of the PTA extracts. A solution of 0.1% PTA did not affect the fibers but did solubilize basal bodies. Running 1.0% PTA extracts from our basal body fractions on sodium dodecyl sulfate (SDS) polyacrylamide gels allowed us to tentatively identify the peptides of basal bodies and kinetodesmal fibers. The latter structures appear to consist of a single 21,000 mol wt peptide. These results also suggest that great caution should be taken in interpreting PTA images, especially of microtubules and axonemes.

scholarly journals On Flagellar Structure in Certain Flagellates

1960 ◽  
Vol 7 (4) ◽  
pp. 697-716 ◽  
Author(s):  
I. R. Gibbons ◽  
A. V. Grimstone
Keyword(s):  
Basal Body ◽  
Basal Bodies ◽  
Complex Structures ◽  
The Third ◽  
Two Systems

This paper describes the structure of the flagella, basal bodies, and some of the associated fibre systems in three genera of complex flagellates, Trichonympha, Pseudotrichonympha, and Holomastigotoides. Three groups of longitudinal fibres occur in a flagellum: two central and nine outer fibres such as have been repeatedly described in other material, and an additional set of nine smaller secondary fibres not previously identified as such. Each central fibre shows a helical substructure; the pair of them are enveloped in a common sheath. Each outer fibre is a doublet with one subfibre bearing projections—called arms—that extend toward the adjacent outer fibre. The basal body is formed by a cylinder of nine triplet outer fibres. Two subfibres of each triplet continue into the flagellum and constitute the doublets. The third subfibre terminates at the transition of basal body to flagellum, possibly giving rise to the nine radial transitional fibres that seem to attach the end of the basal body to the surface of the organism. The central and secondary flagellar fibres are not present in the lumen of the basal body, but other complex structures occur there. The form of these intraluminal structures differs from genus to genus. The flagellar unit is highly asymmetrical. All the flagella examined have possessed the same one of the two possible enantiomorphic forms. At least two systems of fibres are associated with the basal bodies of all three genera.

scholarly journals Cep97 Is Required For Centriole Structural Integrity And Cilia Formation In Drosophila

2019 ◽  
Author(s):  
Jeroen Dobbelaere ◽  
Marketa Schmidt-Cernohorska ◽  
Martina Huranova ◽  
Dea Slade ◽  
Alexander Dammermann
Keyword(s):  
Drosophila Melanogaster ◽  
Basal Body ◽  
Structural Integrity ◽  
Cultured Cells ◽  
Full Length ◽  
Proper Function ◽  
Basal Bodies ◽  
Cell Divisions ◽  
Cilia Formation

SUMMARYCentrioles are highly elaborate microtubule-based structures responsible for the formation of centrosomes and cilia. Despite considerable variation across species and tissues, within any given tissue their size is essentially constant [1, 2]. While the diameter of the centriole cylinder is set by the dimensions of the inner scaffolding structure of the cartwheel [3], how centriole length is set so precisely and stably maintained over many cell divisions is not well understood. Cep97 and CP110 are conserved proteins that localize to the distal end of centrioles and have been reported to limit centriole elongation in vertebrates [4, 5]. Here, we examine Cep97 function in Drosophila melanogaster. We show that Cep97 is essential for formation of full-length centrioles in multiple tissues of the fly. We further identify the microtubule deacetylase Sirt2 as a Cep97 proximity interactor. Deletion of Sirt2 likewise affects centriole size. Interestingly, so does deletion of the acetylase Atat1, indicating that loss of stabilizing acetyl marks impairs centriole integrity. Cep97 and CP110 were originally identified as inhibitors of cilia formation in vertebrate cultured cells [6] and loss of CP110 is a widely used marker of basal body maturation. In contrast, in Drosophila Cep97 is only transiently removed from basal bodies and loss of Cep97 strongly impairs ciliogenesis. Collectively, our results support a model whereby Cep97 functions as part of a protective cap that acts together with the microtubule acetylation machinery to maintain centriole stability, essential for proper function in cilium biogenesis.

scholarly journals ULTRASTRUCTURAL ORGANIZATION OF CILIA AND BASAL BODIES OF THE EPITHELIUM OF THE CHOROID PLEXUS IN THE CHICK EMBRYO

1966 ◽  
Vol 29 (2) ◽  
pp. 333-345 ◽  
Author(s):  
Paul F. Doolin ◽  
Wesley J. Birge
Keyword(s):  
Chick Embryo ◽  
Basal Body ◽  
Osmium Tetroxide ◽  
Basal Plate ◽  
Ultrastructural Organization ◽  
Basal Bodies ◽  
Typical Pattern ◽  
Ultrastructural Studies ◽  
Choroidal Epithelium ◽  
Abnormal Cilia

Ultrastructural studies were performed on normal and abnormal cilia and basal bodies associated with the choroidal epithelium of the chick embryo. Tissues were prepared in each of several fixatives including: 1% osmium tetroxide, in both phosphate and veronal acetate buffers; 2% glutaraldehyde, followed by postfixation in osmium tetroxide; 1% potassium permanganate in veronal acetate buffer. Normal cilia display the typical pattern of 9 peripheral doublets and 2 central fibers, as well as a system of 9 secondary fibers. The latter show distinct interconnections between peripheral and central fibers. Supernumerary fibers were found to occur in certain abnormal cilia. The basal body is complex, bearing 9 transitional fibers at the distal end and numerous cross-striated rootlets at the proximal end. The distal end of the basal body is delimited by a basal plate of moderate density. The tubular cylinder consists of 9 triple fibers. The C subfibers end at the basal plate, whereas subfibers A and B continue into the shaft of the cilium. The 9 transitional fibers radiate out from the distal end of the basal body, ending in bulblike terminal enlargements which are closely associated with the cell membrane in the area of the basal cup. One or 2 prominent basal feet project laterally from the basal body. These structures characteristically show several dense cross-bands and, on occasion, are found associated with microtubules.

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