Inside and outside of the trypanosome flagellum:a multifunctional organelle

Amongst the earliest eukaryotes, trypanosomes have developed conventional organelles but sometimes with extreme features rarely seen in other organisms. This is the case of the flagellum, containing conventional and unique structures whose role in infectivity is still enigmatic.     

The architecture of outer dynein arms in situ

Outer dynein arms, the force generators for axonemal motion, form arrays on microtubule doublets in situ, although they are bouquet-like complexes with separated heads of multiple heavy chains when isolated in vitro. To understand how the three heavy chains are folded in the array, we reconstructed the detailed 3D structure of outer dynein arms of Chlamydomonas flagella in situ by electron cryo-tomography and single-particle averaging. The outer dynein arm binds to the A-microtubule through three interfaces on two adjacent protofilaments, two of which probably represent the docking complex. The three AAA rings of heavy chains, seen as stacked plates, are connected in a striking manner on microtubule doublets. The tail of the alpha-heavy chain, identified by analyzing the oda11 mutant, which lacks alpha-heavy chain, extends from the AAA ring tilted toward the tip of the axoneme and towards the inside of the axoneme at 50 degrees , suggesting a three-dimensional power stroke. The neighboring outer dynein arms are connected through two filamentous structures: one at the exterior of the axoneme and the other through the alpha-tail. Although the beta-tail seems to merge with the alpha-tail at the internal side of the axoneme, the gamma-tail is likely to extend at the exterior of the axoneme and join the AAA ring. This suggests that the fold and function of gamma-heavy chain are different from those of alpha and beta-chains.     

Comparative immunogold analysis of tubulin isoforms in the mouse sperm flagellum: Unique distribution of glutamylated tubulin

The distribution of different tubulin isoforms in the mouse sperm flagellum was studied using four site-directed antibodies to tubulin: DM1A and DM1B general anti α and β-tubulin, 6-11B-1 anti-acetylated α-tubulin, and GT335 anti-glutamylated α and β-tubulin. Quantitative immunogold analyses were performed on five regions of the flagellum: the middle piece, three successive regions of the principal piece, and the terminal piece. A uniform labeling was observed with DM1A and DM1B along the entire flagellum both for peripheral doublets and the central pair. Similar results were obtained with 6-11B-1 directed to acetylated α-tubulin, an N-terminal-modified tubulin isoform. In contrast, the labeling for glutamylated α and β-tubulin, C-terminal modified isoforms, was not uniform. The highest intensity was found in the middle piece and the terminal piece. The labeling which decreased significantly both for peripheral doublets and central pair along the principal piece was considered as a loss of glutamylated tubulin accessibility. From the middle piece to the end of the principal piece, this labeling was predominant in doublets 1-5-6, corresponding to the plane of the flagellar wave. However, the labeling for doublets 2-3-4-7-8-9 was heterogeneous, showing an increasing asymmetry. These results suggest that in the mammalian sperm cell model, the glutamylated tubulin might be involved in a functional heterogeneity among peripheral doublets of the flagellum. © 1996 Wiley-Liss, Inc.     

The axoneme is involved in the elongation of the spermatid nucleus of Drosophila subobscura

Summary

Our analysis of spermiogenesis of Drosophila subobscura indicates that the axoneme takes part in the elongation of the spermatid nucleus, as follows. In sperm of D. subobscura the axoneme accompanies the nucleus in its full length up to the acrosome. Before the elongation of the nucleus begins, the centriole contacts the nuclear membrane, and is orientated with its axis to the centre of the spherical nucleus. Later in development, at the beginning of nucleus elongation, the axis of the centriole does no longer point to the centre of the nucleus but is dislocated more to one side of the nucleus. Subsequently, the axoneme which is growing from the centriole, pushes the nucleus which develops a cap-like structure over the anterior end of the centriole. By the continuosly growing axoneme stretching forces are applied to the anterior part of the nucleus. Consequently the elongating nucleus gets a smaller diameter anteriorly than posteriorly. And the longer the total length of the sperm is the longer is the nucleus. During elongation the chromatin shows a network-like structure. Nucleus elongation stops when the chromatin is fully condensed but the axoneme continues to grow. Thereupon the cap is no longer seen, and the anterior part of the nucleus which previously was the cap, forms now a bulge beside the centriole and the axoneme.     

Sperm axoneme: a tale of tubulin posttranslation diversity

Two microtubule-containing structures are assembled during spermiogenesis: a transient manchette and a stable axoneme. Both structures contain microtubules enriched in posttranslationally modified tubulins. Despite the existence of a spectrum of tubulin isotypes postulated by the multi-tubulin hypothesis, further extended by an elaborated array of posttranslational modifications, it is unknown how this diversity influences microtubule function. There is increasing evidence that different alpha beta-tubulin isotypes can affect the structure and function of microtubules. It is also becoming increasingly clear that eukaryotic cells encode other tubulin proteins expressed by the tubulin superfamily: gamma, delta epsilon, zeta eta, and FtsZ have been identified so far. Although the role of gamma-tubulin in the nucleation of microtubule assembly is well established, the function of delta-, epsilon-, zeta-, eta-, and FtsZ-tubulins is less understood. The members of the tubulin superfamilies found in spermatids include the alpha beta-tubulin dimer, in addition to gamma-tubulin in the centrosome, and delta-tubulin in the perinuclear ring region of the mouse spermatid manchette, the centrosome region, and flagellum. Posttranslational modifications in tubulin isotypes are predominant in the C-terminus exposed on the outside surface of the microtubule. This target site may influence the interaction of microtubule-associated proteins, including motor proteins, and therefore determine the functional specificity of tubulin isotypes. It remains to be determined whether other newcomers to the superfamily of tubulins contain sites prone to posttranslational modification.     

家蚕初级精母细胞内四个基体——轴丝复合体(英语)

应用细胞整装制备和穿透电子显微镜技术,在家蚕4龄幼虫睾丸育精囊的初级精母幼胞内检查出四个基体—轴丝复合体结构.呈两对,每对结构是由连结纤维将基体牢固地联结并使其相互成直角排列.生长的复合体呈现出丰满的远心端膨大.在此阶段,轴丝是由9个具有马达蛋白臂的双微管组成,缺乏辐射轴和两个中央单微管.在基体的三联体微管和轴丝的双微管的接合点,观察到连结节.