1,721,011 research outputs found

    Sas-4 colocalizes with the ciliary rootlets of the drosophila sensory organs

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    The Drosophila eye displays peculiar sensory organs of unknown function, the mechanosensory bristles, that are intercalated among the adjacent ommatidia. Like the other Drosophila sensory organs, the mechanosensory bristles consist of a bipolar neuron and two tandemly aligned centrioles, the distal of which nucleates the ciliary axoneme and represents the starting point of the ciliary rootlets. We report here that the centriole associated protein Sas-4 colocalizes with the short ciliary rootlets of the mechanosensory bristles and with the elongated rootlets of chordotonal and olfactory neurons. This finding suggests an unexpected cytoplasmic localization of Sas-4 protein and points to a new underscored role for this protein. Moreover, we observed that the sheath cells associated with the sensory neurons also display two tandemly aligned centrioles but lacks ciliary axonemes, suggesting that the dendrites of the sensory neurons are dispensable for the assembly of aligned centrioles and rootlets

    A transient microtubule-based structure uncovers a new intrinsic asymmetry between the mother centrioles in the early Drosophila spermatocytes

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    Parent centrioles are characterized in most organisms by individual morphological traits and have distinct asymmetries that provide different functional properties. By contrast, mother and daughter centrioles are morphologically undistinguishable during Drosophila male gametogenesis. Here we report the presence of previously unrecognized microtubule-based structures that extend into the peripheral cytoplasm of the Drosophila polar spermatocytes at the onset of the first meiosis and are positive for the typical centriolar protein Sas-4 and for the kinesin-like protein Klp10A. These structures have a short lifespan and are no longer found in early apolar spermatocytes. Remarkably, each polar spermatocyte holds only one microtubule-based structure that is associated with one of the sister centriole pairs and specifically with the mother centriole. These findings reveal an inherent asymmetry between the parent centrioles at the onset of male meiosis and also uncover unexpected functional properties between the mother centrioles of the same cells

    Cell-to-Cell Interactions during Early Drosophila Oogenesis: An Ultrastructural Analysis

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    Drosophila oogenesis requires the subsequent growth of distinct egg chambers each containing a group of sixteen germline cells surrounded by a simple epithelium of follicle cells. The oocyte occupies a posterior position within the germ cells, thus giving a distinct asymmetry to the egg chamber. Although this disposition is critical for the formation of the anterior-posterior axis of the embryo, the interplay between somatic and germ cells during the early stages of oogenesis remains an open question. We uncover by stage 2, when the egg chambers leaved the germarium, some unique spatial interactions between the posterior follicle cells and the oocyte. These interactions are restricted to the surface of the oocyte over the centriole cluster that formed during early oogenesis. Moreover, the posterior follicle cells in front of the oocyte display a convoluted apical membrane with extensive contacts, whereas the other follicle cells have a flat apical surface without obvious surface protrusions. In addition, the germ cells located at the posterior end of the egg chamber have very elongated protrusions that come into contact with each other or with facing follicle cells. These observations point to distinct polarization events during early oogenesis supporting previous molecular data of an inherent asymmetry between the anterior and the posterior regions of the egg chambers

    The male stem cell niche of Drosophila melanogaster: Interactions between the germline stem cells and the hub

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    The Drosophila male stem cell niche is a well characterized structure in which a small cluster of somatic cells send self-renewal signals to neighbouring germ cells. Although the molecular information involved in the stem cell fate have been identified, much less is understand on the mechanisms driving their short-range specific release. Our ultrastructural analysis reveals distinct protrusions of the stem cell plasma membrane that interdigitate with membrane protrusions of the facing hub cells. Some of these protrusions are very elongated and extend into the hub and could correspond to the Mt-Nanotubes. Therefore, the interface between the stem cells and the hub appears more complex than previously reported and the membrane protrusions of the stem cells might represent specialized surface areas involved in the niche-stem cell communication. We also noticed the presence of clathrin-coated vesicles in the germline plasma membrane that might be also involved in delivering information from the hub

    The Microtubule-Depolymerizing Kinesin-13 Klp10A Is Enriched in the Transition Zone of the Ciliary Structures of Drosophila melanogaster

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    The precursor of the flagellar axoneme is already present in the primary spermatocytes of Drosophila melanogaster. During spermatogenesis each primary spermatocyte shows a centriole pair that moves to the cell membrane and organizes an axoneme-based structure, the cilium-like region (CLR). The CLRs persist through the meiotic divisions and are inherited by young spermatids. During spermatid differentiation the ciliary caps elongate giving rise to the sperm axoneme. Mutations in Klp10A, a kinesin-13 of Drosophila, results in defects of centriole/CLR organization in spermatocytes and of ciliary cap assembly in elongating spermatids. Reduced Klp10A expression also results in strong structural defects of sensory type I neurons. We show, here, that this protein displays a peculiar localization during male gametogenesis. The Klp10A signal is first detected at the distal ends of the centrioles when they dock to the plasma membrane of young primary spermatocytes. At the onset of the first meiotic prometaphase, when the CLRs reach their full size, Klp10A is enriched in a distinct narrow area at the distal end of the centrioles and persists in elongating spermatids at the base of the ciliary cap. We conclude that Klp10A could be a core component of the ciliary transition zone in Drosophila

    Early Drosophila Oogenesis: A Tale of Centriolar Asymmetry

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    Among the morphological processes that characterize the early stages of Drosophila oogenesis, the dynamic of the centrioles deserves particular attention. We re-examined the architecture and the distribution of the centrioles within the germarium and early stages of the vitellarium. We found that most of the germ cell centrioles diverge from the canonical model and display notable variations in size. Moreover, duplication events were frequently observed within the germarium in the absence of DNA replication. Finally, we report the presence of an unusually long centriole that is first detected in the cystoblast and is always associated with the developing oocyte. This centriole is directly inherited after the asymmetric division of the germline stem cells and persists during the process of oocyte selection, thus already representing a marker for oocyte identification at the beginning of its formation and during the ensuing developmental stages

    Microtubule organization during the early development of the parthenogenetic egg of the hymenopteran Muscidifurax uniraptor.

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    The origin of the zygotic centrosome is an important step in developmental biology. It is generally thought that sperm at fertilization plays a central role in forming the functional centrosome which subsequently organizes the first mitotic spindle. However, this view is not applicable in the case of parthenogenetic eggs which develop without the sperm contribution. To clarify the problem of the origin of the zygotic centrosome during parthenogenetic development, we studied a hymenopteran, Muscidifurax uniraptor. Antitubulin antibody revealed that after activation several asters assembled in the egg cytoplasm. The number of asters varied in relation to the cell cycle. They became visible from anaphase of the first meiotic division and increased in number as meiosis progressed, reaching a maximum at the first mitosis. From anaphase-telophase of the first mitosis they decreased in number and were no longer found during the third mitotic division. To elucidate the nature of these asters we performed an ultrastructural study with transmission electron microscopy and immunofluorescence with antibodies against anti-γ-tubulin and CP190. In this way we showed the presence in these asters of centrosomal components and centrioles. Our observations suggest that the cytoplasm of Muscidifurax eggs contains a pool of inactive centrosomal precursor proteins becoming able to nucleate microtubules into well-defined asters containing centrioles after activation

    Centriole and centrosome dynamic during the embryonic cell cycles that follow the formation of the cellular blastoderm in Drosophila

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    We have used immunofluorescence and electron microscopy to examine centrosome dynamics during the first postblastodermic mitoses in the Drosophila embryo. The centrosomal material, as recognized by antibodies against CP190 and γ-tubulin, does not show the typical shape changes observed in syncytial embryos, but remains compact throughout mitosis. Centrioles, however, behave as during the syncytial mitoses, with each daughter cell inheriting two separated centrioles at the end of telophase. During interphase in epithelial cells that have a distinct G1 phase, two isolated centrioles are found, suggesting that the separation of sister centrioles is tightly coupled to a mitotic oscillator in both the 'abbreviated' and the 'complete' embryonic division cycles. The centrioles of the Drosophila embryo sharply differed from the sperm basal body, having a cartwheel structure with nine microtubular doublets and a central tubule. This 'immature' centriolar morphology was shown to persist throughout embryonic development, clearly demonstrating that these centrioles are able to replicate despite their apparently neotenic structure

    Structural characterization of procentrioles in Drosophila spermatids

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    Male gametogenesis in insects is unusual in that the centrioles do not duplicate during the second meiosis and the differentiating spermatids inherit only one centriole. Here it is showed that a distinct procentriole is assembled close to the proximal region of the centriole in early S13 spermatids at the onion stage, confirming previous reports of a proximal centriole-like structure at the proximal end of the spermatid centriole. However, the procentrioles of Drosophila spermatids do not behave like true procentrioles, but their development is blocked at an early stage before the assembly of a complete A-tubule set. Therefore, they may represent early frozen stages of procentriole assembly that do not develop further and eventually disappear in late spermatids
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