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Parasites & Vectors Feb 2023Limited data are currently available on protozoan parasites of the genus Sarcocystis that infect their avian hosts within the order Anseriformes (waterfowl). To date, no...
BACKGROUND
Limited data are currently available on protozoan parasites of the genus Sarcocystis that infect their avian hosts within the order Anseriformes (waterfowl). To date, no Sarcocystis species has been recorded in ducks in China.
METHODS
Leg muscles were sampled from 26 domestic ducks (Anas platyrhynchos) in China in 2021. Morphological characteristics of sarcocysts detected in the muscle tissue were described using light microscopy (LM) and transmission electron microscopy (TEM). Genomic DNA was extracted from single sarcocysts obtained from different ducks, and three genetic markers, 18S ribosomal DNA (18S rDNA), 28S ribosomal DNA (28S rDNA) and mitochondrial (mt) cytochrome oxidase subunit 1 (cox1), were amplified and cloned for sequence analyses.
RESULTS
Sarcocysts were observed by LM in only three of the 28 samples (10.7%). These sarcocysts had a thick cyst wall with numerous brush-like villar protrusions (vps) of 3.8-4.3 μm in length (n = 30) on the cyst surface. TEM observation showed that the sarcocysts had lanceolated vps. Each vps narrowed in the stalk and contained a bundle of microtubules that extended into the ground substance. Comparisons of the new sequences with those deposited in GenBank showed that the most similar sequences were those of Sarcocystis halieti in the great cormorant Phalacrocorax carbo and European starling Sturnus vulgaris, and Sarcocystis calchasi in the domestic pigeon (Columba livia) at the 18S rDNA (99.1% identity); Sarcocystis wenzeli from the domestic chicken Gallus gallus at the 28S rDNA (95.9-96.0% identity); and Sarcocystis speeri from the opossum at the mtcox1 (98.2% identity). The new 18S rDNA, 28S rDNA and mitochondrial cox1 sequences shared up to 99.0%, 95.6% and 97.7% identity, respectively, with those of Sarcocystis spp. obtained from Anseriformes avian hosts. Phylogenetic analysis inferred from the sequences of the three genetic markers placed the organism within a group of Sarcocystis spp. obtained from avian or carnivorous intermediate hosts and avian, marsupial or carnivorous definitive hosts. Based on the morphological observation and molecular analyses, the organism found in the Chinese domestic ducks was regarded as a new species and named Sarcocystis platyrhynchosi n. sp.
CONCLUSIONS
Based on morphology and sequence analyses, the microcysts diagnosed in the domestic ducks examined in this study were named as a new species. This is the first record of Sarcocystis spp. from waterfowl in China. Sarcocysts of similar morphology occur frequently in different Anseriformes birds, and the relationships among these species need to be further clarified in future studies using more molecular markers.
Topics: Animals; Sarcocystis; Ducks; Sarcocystosis; Sarcocystidae; Anseriformes; Columbidae; Phylogeny; Genetic Markers; RNA, Ribosomal, 18S; DNA, Ribosomal; Microscopy, Electron, Transmission; Chickens; China
PubMed: 36732867
DOI: 10.1186/s13071-023-05656-w -
BioRxiv : the Preprint Server For... Sep 2023Cells have complex and beautiful structures that are important for their function, but understanding the molecular mechanisms that produce these structures is a...
Cells have complex and beautiful structures that are important for their function, but understanding the molecular mechanisms that produce these structures is a challenging problem due to the gap in size scales between molecular interactions and cellular structures. The giant ciliate is a unicellular model organism whose large size, reproducible structure, and ability to heal wounds and regenerate has historically allowed the formation of structure in a single cell to be addressed using methods of experimental embryology. Such studies have shown that specific cellular structures, such as the oral apparatus, always form in specific regions of the cell, which raises the question: what is the source of positional information within this organism? By analogy with embryonic development, in which localized mRNA is often used to mark position, we asked whether position along the anterior-posterior axis of Stentor might be marked by specific regionalized mRNAs. By physically bisecting cells and conducting half-cell RNA sequencing, we were able to identify sets of messages enriched in either the anterior or posterior half. We repeated this analysis in cells in which a set of longitudinal microtubule bundles running down the whole length of the cell, known as KM-fibers, were disrupted by RNAi of b-tubulin. We found that many messages either lost their regionalized distribution or switched to an opposite distribution, such that anterior-enriched messages in control became posterior-enriched in the RNAi cells, or vice versa. This study indicates that mRNA can be regionalized within a single giant cell and that microtubules may play a role, possibly by serving as tracks for the movement of the messages.
PubMed: 36711710
DOI: 10.1101/2023.01.09.523364 -
International Journal of Molecular... Jan 2023Although the phagocytic activity of macrophages has long been studied, the involvement of microtubules in the process is not well understood. In this study, we improved...
Although the phagocytic activity of macrophages has long been studied, the involvement of microtubules in the process is not well understood. In this study, we improved the fixation protocol and revealed a dynamically rearranging microtubule network in macrophages, consisting of a basal meshwork, thick bundles at the cell edge, and astral microtubules. Some astral microtubules extended beneath the cell cortex and continued to form bundles at the cell edge. These microtubule assemblies were mutually exclusive of actin accumulation during membrane ruffling. Although the stabilization of microtubules with paclitaxel did not affect the resting stage of the macrophages, it reduced the phagocytic activity and membrane ruffling of macrophages activated with serum-MAF, which induced rapid phagocytosis. In contrast, the destabilization of microtubules with nocodazole enhanced membrane ruffling and the internalization of phagocytic targets suggesting an inhibitory effect of the microtubule network on the remodeling of the actin network. Meanwhile, the microtubule network was necessary for phagosome maturation. Our detailed analyses of cytoskeletal filaments suggest a phagocytosis control system involving Ca influx, the destabilization of microtubules, and activation of actin network remodeling, followed by the translocation and acidification of phagosomes on the microtubule bundles.
Topics: Actins; Phagocytosis; Macrophages; Microtubules; Cytoskeleton
PubMed: 36674886
DOI: 10.3390/ijms24021373 -
Physical Review. X 2022Mixtures of filaments and molecular motors form active materials with diverse dynamical behaviors that vary based on their constituents' molecular properties. To develop...
Mixtures of filaments and molecular motors form active materials with diverse dynamical behaviors that vary based on their constituents' molecular properties. To develop a multiscale of these materials, we map the nonequilibrium phase diagram of microtubules and tip-accumulating kinesin-4 molecular motors. We find that kinesin-4 can drive either global contractions or turbulentlike extensile dynamics, depending on the concentrations of both microtubules and a bundling agent. We also observe a range of spatially heterogeneous nonequilibrium phases, including finite-sized radial asters, 1D wormlike chains, extended 2D bilayers, and system-spanning 3D active foams. Finally, we describe intricate kinetic pathways that yield microphase separated structures and arise from the inherent frustration between the orientational order of filamentous microtubules and the positional order of tip-accumulating molecular motors. Our work reveals a range of novel active states. It also shows that the form of active stresses is not solely dictated by the properties of individual motors and filaments, but is also contingent on the constituent concentrations and spatial arrangement of motors on the filaments.
PubMed: 36643940
DOI: 10.1103/physrevx.12.031006 -
Current Biology : CB Feb 2023Micronuclei resulting from improper chromosome segregation foster chromosome rearrangements. To prevent micronuclei formation in mitosis, the dynamic plus ends of...
Micronuclei resulting from improper chromosome segregation foster chromosome rearrangements. To prevent micronuclei formation in mitosis, the dynamic plus ends of bundled kinetochore microtubules (k-fibers) must establish bipolar attachment with all sister kinetochores on chromosomes, whereas k-fiber minus ends must be clustered at the two opposing spindle poles, which are normally connected with centrosomes. The establishment of chromosome biorientation via k-fiber plus ends is carefully monitored by the spindle assembly checkpoint (SAC). However, how k-fiber minus-end clustering near centrosomes is maintained and monitored remains poorly understood. Here, we show that degradation of NuMA by auxin-inducible degron technologies results in micronuclei formation through k-fiber minus-end detachment from spindle poles during metaphase in HCT116 colon cancer cells. Importantly, k-fiber minus-end detachment from one pole creates misaligned chromosomes that maintain chromosome biorientation and satisfy the SAC, resulting in abnormal chromosome segregation. NuMA depletion also causes minus-end clustering defects in non-transformed Rpe1 cells, but it additionally induces centrosome detachment from partially focused poles, resulting in highly disorganized anaphase. Moreover, we find that NuMA depletion causes centrosome clustering defects in tetraploid-like cells, leading to an increased frequency of multipolar divisions. Together, our data indicate that NuMA is required for faithful chromosome segregation in human mitotic cells, generally by maintaining k-fiber minus-end clustering but also by promoting spindle pole-centrosome or centrosome-centrosome connection in specific cell types or contexts. Similar to erroneous merotelic kinetochore attachments, detachment of k-fiber minus ends from spindle poles evades spindle checkpoint surveillance and may therefore be a source of genomic instability in dividing cells.
Topics: Humans; Centrosome; Chromosome Segregation; Kinetochores; Microtubules; Mitosis; Spindle Apparatus; Spindle Poles
PubMed: 36626904
DOI: 10.1016/j.cub.2022.12.017 -
Plants (Basel, Switzerland) Dec 2022Anticlinal ordinary epidermal cell wall waviness is a widespread feature found in the leaves of a variety of land plant species. However, it has not yet been encountered...
Anticlinal ordinary epidermal cell wall waviness is a widespread feature found in the leaves of a variety of land plant species. However, it has not yet been encountered in leaves with multiple epidermides. Surprisingly, in leaves, ordinary epidermal cells in both layers of the bi-layered adaxial epidermis exhibit wavy anticlinal contour. During the development of the above cells, cortical microtubules are organized in anticlinally oriented bundles under the anticlinal walls, and radial arrays extending from the bundles at the edges of anticlinal and external periclinal walls, under the external periclinal walls. This microtubule pattern is followed by cell wall reinforcement with local thickenings, the cellulose microfibrils of which are parallel to the underlying microtubules. This specialized microtubule organization and concomitant cell wall reinforcement is initiated in the external epidermal layer, while hypodermis follows. The waviness pattern of each epidermal layer is unrelated to that of the other. The above findings are discussed in terms of morphogenetic mechanism induction and any implications in the functional significance of ordinary epidermal cell waviness.
PubMed: 36559549
DOI: 10.3390/plants11243437 -
Brain Research Bulletin Feb 2023Axons are the narrow, up-to-meter long cellular processes of neurons that form the biological cables wiring our nervous system. Most axons must survive for an organism's... (Review)
Review
Axons are the narrow, up-to-meter long cellular processes of neurons that form the biological cables wiring our nervous system. Most axons must survive for an organism's lifetime, i.e. up to a century in humans. Axonal maintenance depends on loose bundles of microtubules that run without interruption all along axons. The continued turn-over and the extension of microtubule bundles during developmental, regenerative or plastic growth requires the availability of α/β-tubulin heterodimers up to a meter away from the cell body. The underlying regulation in axons is poorly understood and hardly features in past and contemporary research. Here we discuss potential mechanisms, particularly focussing on the possibility of local tubulin biogenesis in axons. Current knowledge might suggest that local translation of tubulin takes place in axons, but far less is known about the post-translational machinery of tubulin biogenesis involving three chaperone complexes: prefoldin, CCT and TBC. We discuss functional understanding of these chaperones from a range of model organisms including yeast, plants, flies and mice, and explain what is known from human diseases. Microtubules across species depend on these chaperones, and they are clearly required in the nervous system. However, most chaperones display a high degree of functional pleiotropy, partly through independent functions of individual subunits outside their complexes, thus posing a challenge to experimental studies. Notably, we found hardly any studies that investigate their presence and function particularly in axons, thus highlighting an important gap in our understanding of axon biology and pathology.
Topics: Animals; Humans; Mice; Axons; Microtubules; Neurons; Tubulin
PubMed: 36535305
DOI: 10.1016/j.brainresbull.2022.12.005 -
ILEE: Algorithms and toolbox for unguided and accurate quantitative analysis of cytoskeletal images.The Journal of Cell Biology Feb 2023The eukaryotic cytoskeleton plays essential roles in cell signaling and trafficking, broadly associated with immunity and diseases in humans and plants. To date, most...
The eukaryotic cytoskeleton plays essential roles in cell signaling and trafficking, broadly associated with immunity and diseases in humans and plants. To date, most studies describing cytoskeleton dynamics and function rely on qualitative/quantitative analyses of cytoskeletal images. While state-of-the-art, these approaches face general challenges: the diversity among filaments causes considerable inaccuracy, and the widely adopted image projection leads to bias and information loss. To solve these issues, we developed the Implicit Laplacian of Enhanced Edge (ILEE), an unguided, high-performance approach for 2D/3D-based quantification of cytoskeletal status and organization. Using ILEE, we constructed a Python library to enable automated cytoskeletal image analysis, providing biologically interpretable indices measuring the density, bundling, segmentation, branching, and directionality of the cytoskeleton. Our data demonstrated that ILEE resolves the defects of traditional approaches, enables the detection of novel cytoskeletal features, and yields data with superior accuracy, stability, and robustness. The ILEE toolbox is available for public use through PyPI and Google Colab.
Topics: Humans; Algorithms; Image Processing, Computer-Assisted; Microtubules; Cytoskeleton; Plants; Animals
PubMed: 36534166
DOI: 10.1083/jcb.202203024 -
ACS Omega Dec 2022Active networks of biopolymers and motor proteins in vitro self-organize and exhibit dynamic structures on length scales much larger than the interacting individual...
Active networks of biopolymers and motor proteins in vitro self-organize and exhibit dynamic structures on length scales much larger than the interacting individual components of which they consist. How the dynamics is related across the range of length scales is still an open question. Here, we experimentally characterize and quantify the dynamic behavior of isolated microtubule bundles that bend due to the activity of motor proteins. At the motor level, we track and describe the motion features of kinesin-1 clusters stepping within the bending bundles. We find that there is a separation of length scales by at least 1 order of magnitude. At a run length of <1 μm, kinesin-1 activity leads to a bundle curvature in the range of tens of micrometers. We propose that the distribution of microtubule polarity plays a crucial role in the bending dynamics that we observe at both the bundle and motor levels. Our results contribute to the understanding of fundamental principles of vital intracellular processes by disentangling the multiscale dynamics in out-of-equilibrium active networks composed of cytoskeletal elements.
PubMed: 36506136
DOI: 10.1021/acsomega.2c04958 -
Proceedings of the National Academy of... Dec 2022Cellular morphogenesis and processes such as cell division and migration require the coordination of the microtubule and actin cytoskeletons. Microtubule-actin crosstalk...
Cellular morphogenesis and processes such as cell division and migration require the coordination of the microtubule and actin cytoskeletons. Microtubule-actin crosstalk is poorly understood and largely regarded as the capture and regulation of microtubules by actin. Septins are filamentous guanosine-5'-triphosphate (GTP) binding proteins, which comprise the fourth component of the cytoskeleton along microtubules, actin, and intermediate filaments. Here, we report that septins mediate microtubule-actin crosstalk by coupling actin polymerization to microtubule lattices. Superresolution and platinum replica electron microscopy (PREM) show that septins localize to overlapping microtubules and actin filaments in the growth cones of neurons and non-neuronal cells. We demonstrate that recombinant septin complexes directly crosslink microtubules and actin filaments into hybrid bundles. In vitro reconstitution assays reveal that microtubule-bound septins capture and align stable actin filaments with microtubules. Strikingly, septins enable the capture and polymerization of growing actin filaments on microtubule lattices. In neuronal growth cones, septins are required for the maintenance of the peripheral actin network that fans out from microtubules. These findings show that septins directly mediate microtubule interactions with actin filaments, and reveal a mechanism of microtubule-templated actin growth with broader significance for the self-organization of the cytoskeleton and cellular morphogenesis.
Topics: Septins; Actins; Microtubules
PubMed: 36475946
DOI: 10.1073/pnas.2202803119