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https://www.readbyqxmd.com/read/28760733/wasp-arp2-3-dependent-actin-polymerization-influences-fusogen-localization-during-cell-cell-fusion-in-c-elegans-embryos
#1
Yan Zhang, Yihong Yang, Zhiwen Zhu, Guangshuo Ou
Cell-cell fusion is essential for development and physiology. Actin polymerization was implicated in the C. elegans fusogen EFF-1 engagement in a reconstituted Drosophila cell culture system, and the actin-binding protein spectraplakin links EFF-1 to the actin cytoskeleton and promotes cell-cell fusions in C. elegans larvae. However, it remains unclear whether and how fusogens and the actin cytoskeleton are coordinated in C. elegans embryos. Here, we used live imaging analysis of GFP knock-in and RNAi embryos to study the embryonic cell-cell fusions in C...
July 31, 2017: Biology Open
https://www.readbyqxmd.com/read/28759579/dynactin-binding-to-tyrosinated-microtubules-promotes-centrosome-centration-in-c-elegans-by-enhancing-dynein-mediated-organelle-transport
#2
Daniel J Barbosa, Joana Duro, Bram Prevo, Dhanya K Cheerambathur, Ana X Carvalho, Reto Gassmann
The microtubule-based motor dynein generates pulling forces for centrosome centration and mitotic spindle positioning in animal cells. How the essential dynein activator dynactin regulates these functions of the motor is incompletely understood. Here, we dissect the role of dynactin's microtubule binding activity, located in the p150 CAP-Gly domain and an adjacent basic patch, in the C. elegans zygote. Analysis of p150 mutants engineered by genome editing suggests that microtubule tip tracking of dynein-dynactin is dispensable for targeting the motor to the cell cortex and for generating robust cortical pulling forces...
July 2017: PLoS Genetics
https://www.readbyqxmd.com/read/28739679/two-populations-of-cytoplasmic-dynein-contribute-to-spindle-positioning-in-c-elegans-embryos
#3
Ruben Schmidt, Lars-Eric Fielmich, Ilya Grigoriev, Eugene A Katrukha, Anna Akhmanova, Sander van den Heuvel
The position of the mitotic spindle is tightly controlled in animal cells as it determines the plane and orientation of cell division. Contacts between cytoplasmic dynein and astral microtubules (MTs) at the cell cortex generate pulling forces that position the spindle. An evolutionarily conserved Gα-GPR-1/2(Pins/LGN)-LIN-5(Mud/NuMA) cortical complex interacts with dynein and is required for pulling force generation, but the dynamics of this process remain unclear. In this study, by fluorescently labeling endogenous proteins in Caenorhabditis elegans embryos, we show that dynein exists in two distinct cortical populations...
July 24, 2017: Journal of Cell Biology
https://www.readbyqxmd.com/read/28701691/spatiotemporal-expression-profiling-of-long-intervening-noncoding-rnas-in-caenorhabditis-elegans
#4
Weihong Liu, Enchao Yu, Siyu Chen, Xiaopeng Ma, Yiwen Lu, Xiao Liu
To better understand the biological function of long noncoding RNAs, it is critical to determine their spatiotemporal expression patterns. We generated transgenic reporter strains for 149 out of the 170 annotated C. elegans long intervening noncoding RNAs (lincRNAs) and profiled their temporal activity. For the 68 lincRNAs with integrated reporter lines, we profiled their expression at the resolution of single cells in L1 larvae, and revealed that the expression of lincRNAs is more specific, heterogeneous and at lower level than transcription factors (TFs)...
July 12, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28701347/a-casein-kinase-1-prevents-expulsion-of-the-oocyte-meiotic-spindle-into-a-polar-body-by-regulating-cortical-contractility
#5
Jonathan R Flynn, Francis J McNally
During female meiosis, haploid eggs are generated from diploid oocytes. This reduction in chromosome number occurs through two highly asymmetric cell divisions resulting in one large egg and two small polar bodies. Unlike mitosis, where an actomyosin contractile ring forms between the sets of segregating chromosomes, the meiotic contractile ring forms on the cortex adjacent to one spindle pole, then ingresses down the length of the spindle to position itself at the exact midpoint between the two sets of segregating chromosomes...
July 12, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28701343/let-99-functions-in-the-astral-furrowing-pathway-where-it-is-required-for-myosin-enrichment-in-the-contractile-ring
#6
Kari L Price, Lesilee S Rose
The anaphase spindle determines the position of the cytokinesis furrow, such that the contractile ring assembles in an equatorial zone between the two spindle poles. Contractile ring formation is mediated by RhoA activation at the equator by the centralspindlin complex and midzone microtubules. Astral microtubules also inhibit RhoA accumulation at the poles. In the C. elegans one-cell embryo, the astral microtubule dependent pathway requires anillin, NOP-1 and LET-99. LET-99 is well characterized for generating the asymmetric cortical localization of the Gα-dependent force-generating complex that positions the spindle during asymmetric division...
July 12, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28701341/computer-simulations-reveal-mechanisms-that-organize-nuclear-dynein-forces-to-separate-centrosomes
#7
Alessandro De Simone, Pierre Gönczy
Centrosome separation along the surface of the nucleus at the onset of mitosis is critical for bipolar spindle assembly. Dynein anchored on the nuclear envelope is known to be important for centrosome separation, but it is unclear how nuclear dynein forces are organized in an anisotropic manner to promote the movement of centrosomes away from each other. Here, we use computational simulations of C. elegans embryos to address this fundamental question, testing three potential mechanisms by which nuclear dynein may act...
July 12, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28659600/comparative-genetic-proteomic-and-phosphoproteomic-analysis-of-c-elegans-embryos-with-a-focus-on-ham-1-stox-and-pig-1-melk-in-dopaminergic-neuron-development
#8
Sarah-Lena Offenburger, Dalila Bensaddek, Alejandro Brenes Murillo, Angus I Lamond, Anton Gartner
Asymmetric cell divisions are required for cellular diversity and defects can lead to altered daughter cell fates and numbers. In a genetic screen for C. elegans mutants with defects in dopaminergic head neuron specification or differentiation, we isolated a new allele of the transcription factor HAM-1 [HSN (Hermaphrodite-Specific Neurons) Abnormal Migration]. Loss of both HAM-1 and its target, the kinase PIG-1 [PAR-1(I)-like Gene], leads to abnormal dopaminergic head neuron numbers. We identified discrete genetic relationships between ham-1, pig-1 and apoptosis pathway genes in dopaminergic head neurons...
June 28, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28655887/comparative-proteome-analysis-between-c-briggsae-embryos-and-larvae-reveals-a-role-of-chromatin-modification-proteins-in-embryonic-cell-division
#9
Xiaomeng An, Jiaofang Shao, Huoming Zhang, Xiaoliang Ren, Vincy Wing Sze Ho, Runsheng Li, Ming-Kin Wong, Zhongying Zhao
Caenorhabditis briggsae has emerged as a model for comparative biology against model organism C. elegans. Most of its cell fate specifications are completed during embryogenesis whereas its cell growth is achieved mainly in larval stages. The molecular mechanism underlying the drastic developmental changes is poorly understood. To gain insights into the molecular changes between the two stages, we compared the proteomes between the two stages using iTRAQ. We identified a total of 2,791 proteins in the C. briggsae embryos and larvae, 247 of which undergo up- or down-regulation between the two stages...
June 27, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28637841/protein-tyrosine-phosphatase-%C3%AE-mediates-the-sema3a-induced-cortical-basal-dendritic-arborization-through-the-activation-of-fyn-tyrosine-kinase
#10
Fumio Nakamura, Takako Okada, Maria Shishikura, Noriko Uetani, Masahiko Taniguchi, Takeshi Yagi, Yoichiro Iwakura, Toshio Ohshima, Yoshio Goshima, Stephen M Strittmatter
Leukocyte common antigen-related (LAR) class protein tyrosine phosphatases (PTPs) are critical for axonal guidance; however, their relation to specific guidance cues is poorly defined. We here show that PTP-3, a LAR homolog in Caenorhabditis elegans, is involved in axon guidance regulated by Semaphorin-2A-signaling. PTPδ, one of the vertebrate LAR class PTPs, participates in the Semaphorin-3A (Sema3A)-induced growth cone collapse response of primary cultured dorsal root ganglion neurons from Mus musculus embryos...
July 26, 2017: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
https://www.readbyqxmd.com/read/28637747/f-actin-prevents-interaction-between-sperm-dna-and-the-oocyte-meiotic-spindle-in-c-elegans
#11
Michelle T Panzica, Harold C Marin, Anne-Cecile Reymann, Francis J McNally
Fertilization occurs during female meiosis in most animals, which raises the question of what prevents the sperm DNA from interacting with the meiotic spindle. In this study, we find that Caenorhabditis elegans sperm DNA stays in a fixed position at the opposite end of the embryo from the meiotic spindle while yolk granules are transported throughout the embryo by kinesin-1. In the absence of F-actin, the sperm DNA, centrioles, and organelles were transported as a unit with the yolk granules, resulting in sperm DNA within 2 µm of the meiotic spindle...
August 7, 2017: Journal of Cell Biology
https://www.readbyqxmd.com/read/28634273/the-c-elegans-vapb-homolog-vpr-1-is-a-permissive-signal-for-gonad-development
#12
Pauline A Cottee, Tim Cole, Jessica Schultz, Hieu D Hoang, Jack Vibbert, Sung Min Han, Michael A Miller
VAMP/synaptobrevin-associated proteins (VAPs) contain an N-terminal major sperm protein domain (MSPd) that is associated with amyotrophic lateral sclerosis. VAPs have an intracellular housekeeping function, as well as an extracellular signaling function mediated by the secreted MSPd. Here we show that the C. elegans VAP homolog VPR-1 is essential for gonad development. vpr-1 null mutants are maternal effect sterile due to arrested gonadogenesis following embryo hatching. Somatic gonadal precursor cells and germ cells fail to proliferate fully and complete their respective differentiation programs...
June 15, 2017: Development
https://www.readbyqxmd.com/read/28630487/multicolor-two-photon-imaging-of-endogenous-fluorophores-in-living-tissues-by-wavelength-mixing
#13
Chiara Stringari, Lamiae Abdeladim, Guy Malkinson, Pierre Mahou, Xavier Solinas, Isabelle Lamarre, Sébastien Brizion, Jean-Baptiste Galey, Willy Supatto, Renaud Legouis, Ana-Maria Pena, Emmanuel Beaurepaire
Two-photon imaging of endogenous fluorescence can provide physiological and metabolic information from intact tissues. However, simultaneous imaging of multiple intrinsic fluorophores, such as nicotinamide adenine dinucleotide(phosphate) (NAD(P)H), flavin adenine dinucleotide (FAD) and retinoids in living systems is generally hampered by sequential multi-wavelength excitation resulting in motion artifacts. Here, we report on efficient and simultaneous multicolor two-photon excitation of endogenous fluorophores with absorption spectra spanning the 750-1040 nm range, using wavelength mixing...
June 19, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28629355/biologically-constrained-optimization-based-cell-membrane-segmentation-in-c-elegans-embryos
#14
Yusuke Azuma, Shuichi Onami
BACKGROUND: Recent advances in bioimaging and automated analysis methods have enabled the large-scale systematic analysis of cellular dynamics during the embryonic development of Caenorhabditis elegans. Most of these analyses have focused on cell lineage tracing rather than cell shape dynamics. Cell shape analysis requires cell membrane segmentation, which is challenging because of insufficient resolution and image quality. This problem is currently solved by complicated segmentation methods requiring laborious and time consuming parameter adjustments...
June 19, 2017: BMC Bioinformatics
https://www.readbyqxmd.com/read/28623274/reduction-in-chromosome-mobility-accompanies-nuclear-organization-during-early-embryogenesis-in-caenorhabditis-elegans
#15
Ritsuko Arai, Takeshi Sugawara, Yuko Sato, Yohei Minakuchi, Atsushi Toyoda, Kentaro Nabeshima, Hiroshi Kimura, Akatsuki Kimura
In differentiated cells, chromosomes are packed inside the cell nucleus in an organised fashion. In contrast, little is known about how chromosomes are packed in undifferentiated cells and how nuclear organization changes during development. To assess changes in nuclear organization during the earliest stages of development, we quantified the mobility of a pair of homologous chromosomal loci in the interphase nuclei of Caenorhabditis elegans embryos. The distribution of distances between homologous loci was consistent with a random distribution up to the 8-cell stage but not at later stages...
June 16, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28619826/a-toolkit-for-gfp-mediated-tissue-specific-protein-degradation-in-c-elegans
#16
Shaohe Wang, Ngang Heok Tang, Pablo Lara-Gonzalez, Zhiling Zhao, Dhanya K Cheerambathur, Bram Prevo, Andrew D Chisholm, Arshad Desai, Karen Oegema
Proteins that are essential for embryo production, cell division and early embryonic events are frequently reused later in embryogenesis, during organismal development or in the adult. Examining protein function across these different biological contexts requires tissue-specific perturbation. Here, we describe a method that uses expression of a fusion between a GFP-targeting nanobody and a SOCS-box containing ubiquitin ligase adaptor to target GFP-tagged proteins for degradation. When combined with endogenous locus GFP tagging by CRISPR-Cas9 or with rescue of a null mutant with a GFP fusion, this approach enables routine and efficient tissue-specific protein ablation...
July 15, 2017: Development
https://www.readbyqxmd.com/read/28502614/partially-compromised-specification-causes-stochastic-effects-on-gut-development-in-c-elegans
#17
Hailey Choi, Gina Broitman-Maduro, Morris F Maduro
The C. elegans gut descends from the E progenitor cell through a series of stereotyped cell divisions and morphogenetic events. Effects of perturbations of upstream cell specification on downstream organogenesis have not been extensively investigated. Here we have assembled an allelic series of strains that variably compromise specification of E by perturbing the activation of the gut-specifying end-1 and end-3 genes. Using a marker that allows identification of all E descendants regardless of fate, superimposed with markers that identify cells that have adopted a gut fate, we have examined the fate of E lineage descendants among hundreds of embryos...
May 11, 2017: Developmental Biology
https://www.readbyqxmd.com/read/28500339/a-role-for-core-planar-polarity-proteins-in-cell-contact-mediated-orientation-of-planar-cell-division-across-the-mammalian-embryonic-skin
#18
Fazal Oozeer, Laura L Yates, Charlotte Dean, Caroline J Formstone
The question of how cell division orientation is determined is fundamentally important for understanding tissue and organ shape in both healthy or disease conditions. Here we provide evidence for cell contact-dependent orientation of planar cell division in the mammalian embryonic skin. We propose a model where the core planar polarity proteins Celsr1 and Frizzled-6 (Fz6) communicate the long axis orientation of interphase basal cells to neighbouring basal mitoses so that they align their horizontal division plane along the same axis...
May 12, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28492281/c-elegans-chromosomes-connect-to-centrosomes-by-anchoring-into-the-spindle-network
#19
Stefanie Redemann, Johannes Baumgart, Norbert Lindow, Michael Shelley, Ehssan Nazockdast, Andrea Kratz, Steffen Prohaska, Jan Brugués, Sebastian Fürthauer, Thomas Müller-Reichert
The mitotic spindle ensures the faithful segregation of chromosomes. Here we combine the first large-scale serial electron tomography of whole mitotic spindles in early C. elegans embryos with live-cell imaging to reconstruct all microtubules in 3D and identify their plus- and minus-ends. We classify them as kinetochore (KMTs), spindle (SMTs) or astral microtubules (AMTs) according to their positions, and quantify distinct properties of each class. While our light microscopy and mutant studies show that microtubules are nucleated from the centrosomes, we find only a few KMTs directly connected to the centrosomes...
May 11, 2017: Nature Communications
https://www.readbyqxmd.com/read/28482037/a-non-canonical-site-reveals-the-cooperative-mechanisms-of-microrna-mediated-silencing
#20
Mathieu N Flamand, Hin Hark Gan, Vinay K Mayya, Kristin C Gunsalus, Thomas F Duchaine
Although strong evidence supports the importance of their cooperative interactions, microRNA (miRNA)-binding sites are still largely investigated as functionally independent regulatory units. Here, a survey of alternative 3΄UTR isoforms implicates a non-canonical seedless site in cooperative miRNA-mediated silencing. While required for target mRNA deadenylation and silencing, this site is not sufficient on its own to physically recruit miRISC. Instead, it relies on facilitating interactions with a nearby canonical seed-pairing site to recruit the Argonaute complexes...
May 8, 2017: Nucleic Acids Research
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