Cytoskeletal discontinuities in the cell body cortex initiate basal body assembly and oral development in the ciliate Stentor

Development ◽  
1985 ◽  
Vol 87 (1) ◽  
pp. 249-257
Author(s):  
Nöel De Terra

My previous work has shown that disconnecting the oral apparatus of Stentor into two parts induces mass assembly of basal bodies on the ventral cell surface and thus initiates oral development. This operation severs the extensive microtubule tracts joining the oral membranelles at their bases. To determine whether basal body assembly and oral development are also induced by permanently disconnecting the longitudinal microtubule fibre tracts (mt fibre tracts) of the cell body cortex, I interposed a ring of inverted (heteropolar) cortex between the anterior and posterior halves of interphase stentors. When successful, this operation made it impossible for these fibre tracts to rejoin at the heteropolar boundaries and always induced basal body assembly and oral development in the graft complex. By contrast, tripartite homopolar graft complexes rarely initiated oral development; when they did, it was apparently in response to the presence of disproportionately small oral structures, which is the normal stimulus for oral development in Stentor. The mt fibre tracts of tripartite homopolar grafts also eventually became continuous. These results support the hypothesis that permanent, extensive discontinuities anywhere within the cortical cytoskeleton can trigger basal body assembly and oral development. Since the onset of these processes is known to initiate cell division in Stentor, the results also suggest that development of discontinuities within the cortical cytoskeleton during interphase growth may be the endogenous stimulus initiating cell division in Stentor.

Development ◽  
1985 ◽  
Vol 87 (1) ◽  
pp. 241-247
Author(s):  
Nöel De Terra

In the ciliate Stentor, many thousands of basal bodies assemble on the ventral cell surface to form a new oral apparatus during cell division, regeneration and reorganization (oral replacement during interphase). During interphase, oral development is normally inhibited by the presence of the anteriorly placed oral apparatus. A glass needle was used to cut the oral apparatus of interphase stentors in two so that the parts remained intact but separate at the anterior end of the cell. These cells initiated basal body assembly and oral development, usually within 8 h. Basal body assembly can therefore result from disconnection of fibrous structures within the oral apparatus but is unlikely to be regulated by an inhibitor diffusing from it.


2013 ◽  
Vol 24 (9) ◽  
pp. 1321-1333 ◽  
Author(s):  
Ana Lozano-Núñez ◽  
Kyojiro N. Ikeda ◽  
Thomas Sauer ◽  
Christopher L. de Graffenried

Polo-like kinases are important regulators of cell division, playing diverse roles in mitosis and cytoskeletal inheritance. In the parasite Trypanosoma brucei, the single PLK homologue TbPLK is necessary for the assembly of a series of essential organelles that position and adhere the flagellum to the cell surface. Previous work relied on RNA interference or inhibitors of undefined specificity to inhibit TbPLK, both of which have significant experimental limitations. Here we use an analogue-sensitive approach to selectively and acutely inhibit TbPLK. T. brucei cells expressing only analogue-sensitive TbPLK (TbPLKas) grow normally, but upon treatment with inhibitor develop defects in flagellar attachment and cytokinesis. TbPLK cannot migrate effectively when inhibited and remains trapped in the posterior of the cell throughout the cell cycle. Using synchronized cells, we show that active TbPLK is a direct requirement for the assembly and extension of the flagellum attachment zone, which adheres the flagellum to the cell surface, and for the rotation of the duplicated basal bodies, which positions the new flagellum so that it can extend without impinging on the old flagellum. This approach should be applicable to the many kinases found in the T. brucei genome that lack an ascribed function.


Development ◽  
1998 ◽  
Vol 125 (7) ◽  
pp. 1305-1314
Author(s):  
M. Jerka-Dziadosz ◽  
F. Ruiz ◽  
F. Beisson

In Paramecium the development of cell shape and surface pattern during division depends on a precise spatial and temporal pattern of duplication of the ciliary basal bodies which are the organizers of the cortical cytoskeleton. According to their localization, basal bodies will duplicate once, more than once or not all and this duplication is coupled with cell division, as is centrosomal duplication in metazoan cells. We describe here a monogenic nuclear recessive mutation, crochu1 (cro1), resulting in abnormal cell shape and cortical pattern and hypersensitivity to nocodazole. The cytological analysis, by immunofluorescence and electron microscopy, demonstrates that the mutation causes hyper duplication of basal bodies and releases both spatial and temporal control of duplication as basal bodies continue to proliferate in interphase and do so at ectopic locations, beneath the surface and in cortical territories where no duplication occurs in the wild type. However, the abnormal surface organization of cro1 cells does not affect the program of basal body duplication during division. By genetic analysis, no interaction was detected with the sm19 mutation which impairs basal body duplication. In contrast, the cro1 mutation suppresses the nocodazole resistance conferred by nocr1, a mutation in a beta-tubulin gene. This interaction suggests that the primary effect of the mutation bears on microtubule dynamics, whose instability, normally increased during division, would persist throughout the interphase and provide a signal for constitutive basal body duplication.


1978 ◽  
Vol 31 (1) ◽  
pp. 25-35
Author(s):  
M.A. Gillott ◽  
R.E. Triemer

The ultrastructure of mitosis in Euglena gracilis was investigated. At preprophase the nucleus migrates anteriorly and associates with the basal bodies. Flagella and basal bodies replicate at preprophase. Cells retain motility throughout division. The reservoir and the prophase nucleus elongate perpendicular to the incipient cleavage furrow. One basal body pair surrounded by a ribosome-free zone is found at each of the nuclear poles. The spindle forms within the intact nuclear envelope- Polar fenestrae are absent. At metaphase, the endosome is elongated from pole to pole, and chromosomes are loosely arranged in the equatorial region. Distinct, trilayered kinetochores are present. Spindle elongates as chromosomes migrate to the poles forming a dumb-bell shaped nucleus by telophase. Daughter nuclei are formed by constriction of the nuclear envelope. Cytokinesis is accomplished by furrowing. Cell division in Euglena is compared with that of certain other algae.


We have analysed the timing and order of events occurring within the cell division cycle of Trypanosoma brucei . Cells in the earliest stages of the cell cycle possess a single copy of three major organelles: the nucleus, the kinetoplast and the flagellum. The first indication of progress through the cell cycle is the elongation of the pro-basal body lying adjacent to the mature basal body subtending the flagellum. This newly elongated basal body occupies a posterior position within the cell when it initiates growth of the new daughter flagellum. Genesis of two new pro-basal bodies occurs only after growth of the new daughter flagellum has been initiated. Extension of the new flagellum, together with the paraflagellar rod, then continues throughout a major portion of the cell cycle. During this period of flagellum elongation, kinetoplast division occurs and the two kinetoplasts, together with the two flagellar basal bodies, then move apart within the cell. Mitosis is then initiated and a complex pattern of organelle positions is achieved whereby a division plane runs longitudinally through the cell such that each daughter ultimately receives a single nucleus, kinetoplast and flagellum. These events have been described from observations of whole cytoskeletons by transmission electron microscopy together with detection of particular organelles by fluorescence microscopy. The order and timing of events within the cell cycle has been derived from analyses of the proportion of a given cell type occurring within an exponentially growing culture.


1982 ◽  
Vol 60 (10) ◽  
pp. 2296-2308 ◽  
Author(s):  
Donald Jones ◽  
James D. Berger

Nine temperature-sensitive gene mutations affecting cellular morphogenesis were analysed and shown to be single recessive genes. Their phenotypes fall into three classes: small mutants (sm) which interfere with cell surface and basal body proliferation to produce short cells; defective fission zone mutants (dfz) which do not form a complete fission zone during cell division; and defective constriction mutants (dc) which form a normal fission zone, but do not constrict properly. In sm2 cells there is a reduction in the number of basal bodies and in the amount of cell surface produced preceding fission. This results in the production of truncated daughter cells in which most of the normal structures of either the anterior or posterior part of the cell are highly reduced or missing. Production of basal bodies in gullet primordia is also abnormal. The dfz mutants act early in the fission process to block the formation of the fission zone which precedes the formation of the fission furrow. The dc mutations act later in the fission process and lead to failure of daughter cell separation. One mutant, dc3, also shows slightly reduced proliferation of cell surface. This defect occurs prior to fission.


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 .


1977 ◽  
Vol 25 (1) ◽  
pp. 367-385
Author(s):  
R.K. Peck

The ultrastructure of the somatic cortex of the ciliate Pseudomicrothorax dubius is studied with emphasis on the epiplasm layer which lies immediately under the inner alveolar membrane and is continuous with the terminal plates of cortical basal bodies. In addition to a clearly demonstrable cytoskeletal role, the epiplasm appears to function as a comenting substance which integrates numerous cortical fibres and membranes. The kinetodesmal, postciliary and transverse fibre systems which originate at the proximal ends of basal bodies extend toward the cell surface and end at or in the epiplasm. Inner alveolar membranes and trichocyst membranes are attached to the epiplasm. Basal bodies are anchored into the epiplasm via their terminal plates. The epiplasm appears to be morphogenetically important as a matrix into which newly formed basal bodies insert. Electron-opaque arms occur at the terminal plate level of new basal bodies, and these arms fuse with the epiplasm when basal body insertion occurs. The position of trichocysts in the cortex is specified by the epiplasm. Evidence from numerous other ciliates tends to confirm both structural and morphogenetic roles of the epiplasm.


1961 ◽  
Vol 11 (1) ◽  
pp. 179-205 ◽  
Author(s):  
I. R. Gibbons

This paper describes the fine structure and its relationship to the direction of beat in four types of cilia on the gill of the fresh-water mussel Anodonta cataracta. The cilia contain nine outer, nine secondary, and two central fibers, such as have been described previously in other material. Each outer fiber is a doublet with one subfiber bearing arms. One particular pair of outer fibers (numbers 5 and 6) are joined together by a bridge. The two central fibers are enclosed by a central sheath; also present in this region is a single, small mid-fiber. The different groups of fibers are connected together by radial links that extend from the outer to the secondary fibers, and from the secondary fibers to the central sheath. The basal body consists of a cylinder of nine triplet fibers. Projecting from it on one side is a dense conical structure called the basal foot. The cylinder of outer fibers continues from the basal body into the cilium, passing through a complex transitional region in which five distinct changes of structure occur at different levels. There are two sets of fibers associated with the basal bodies: a pair of striated rootlets that extends from each basal body down into the cell, and a system of fine tubular fibers that runs parallel to the cell surface. The relationship between fine structure and direction of beat is the same in all four types of cilia examined. The plane of beat is perpendicular to the plane of the central fibers, with the effective stroke toward the bridge between outer fibers 5 and 6, and toward the foot on the basal body.


1966 ◽  
Vol 31 (1) ◽  
pp. 43-54 ◽  
Author(s):  
Allan D. Dingle ◽  
Chandler Fulton

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.


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