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https://www.readbyqxmd.com/read/29776969/identifying-genetic-players-in-cell-sheet-morphogenesis-using-a-drosophila-deficiency-screen-for-genes-on-chromosome-2r-involved-in-dorsal-closure
#1
Richard D Mortensen, Regan P Moore, Stephanie M Fogerson, Hellen Y Chiou, Chimdindu V Obinero, Neel K Prabhu, Angela H Wei, Janice M Crawford, Daniel P Kiehart
Cell sheet morphogenesis characterizes key developmental transitions and homeostasis, in vertebrates and throughout phylogeny, including gastrulation, neural tube formation and wound healing. Dorsal closure, a process during Drosophila embryogenesis, has emerged as a model for cell sheet morphogenesis. ∼140 genes are currently known to affect dorsal closure and new genes are identified each year. Many of these genes were identified in screens that resulted in arrested development. Dorsal closure is remarkably robust and many questions regarding the molecular mechanisms involved in this complex biological process remain...
May 18, 2018: G3: Genes—Genomes—Genetics
https://www.readbyqxmd.com/read/29773790/crispr-cas9-mediated-gene-targeting-in-arabidopsis-using-sequential-transformation
#2
Daisuke Miki, Wenxin Zhang, Wenjie Zeng, Zhengyan Feng, Jian-Kang Zhu
Homologous recombination-based gene targeting is a powerful tool for precise genome modification and has been widely used in organisms ranging from yeast to higher organisms such as Drosophila and mouse. However, gene targeting in higher plants, including the most widely used model plant Arabidopsis thaliana, remains challenging. Here we report a sequential transformation method for gene targeting in Arabidopsis. We find that parental lines expressing the bacterial endonuclease Cas9 from the egg cell- and early embryo-specific DD45 gene promoter can improve the frequency of single-guide RNA-targeted gene knock-ins and sequence replacements via homologous recombination at several endogenous sites in the Arabidopsis genome...
May 17, 2018: Nature Communications
https://www.readbyqxmd.com/read/29773559/serrate-notch-signaling-regulates-the-size-of-the-progenitor-cell-pool-in-drosophila-imaginal-rings
#3
Sheng-An Yang, Wu-Min Deng
Drosophila imaginal rings are larval tissues composed of progenitor cells that are essential for the formation of adult foreguts, hindguts and salivary glands. Specified from subsets of ectoderm in the embryo, imaginal ring cells are kept quiescent until mid-second larval instar, and undergo rapid proliferation during the third instar to attain adequate numbers of cells that will replace apoptotic larval tissues for adult organ formation. Here we show that Notch signaling is activated in all three imaginal rings from middle embryonic stage to early pupal stage and that Notch signaling positively controls cell proliferation in all three imaginal rings during the third larval instar...
May 17, 2018: Genetics
https://www.readbyqxmd.com/read/29773558/genetic-analyses-of-elys-mutations-in-drosophila-show-maternal-effect-lethality-and-interactions-with-nucleoporin-genes
#4
Kazuyuki Hirai, Zhuo Wang, Kohei Miura, Takaaki Hayashi, Takeshi Awasaki, Moe Wada, Yoko Keira, Hiroyuki O Ishikawa, Kyoichi Sawamura
ELYS determines the subcellular localizations of Nucleoporins (Nups) during interphase and mitosis. We made loss-of-function mutations of Elys in Drosophila melanogaster and found that ELYS is dispensable for zygotic viability and male fertility but the maternal supply is necessary for embryonic development. Subsequent to fertilization, mitotic progression of the embryos produced by the mutant females is severely disrupted at the first cleavage division, accompanied by irregular behavior of mitotic centrosomes...
May 18, 2018: G3: Genes—Genomes—Genetics
https://www.readbyqxmd.com/read/29764918/a-feed-forward-relay-integrates-the-regulatory-activities-of-bicoid-and-orthodenticle-via-sequential-binding-to-suboptimal-sites
#5
Rhea R Datta, Jia Ling, Jesse Kurland, Xiaotong Ren, Zhe Xu, Gozde Yucel, Jackie Moore, Leila Shokri, Isabel Baker, Timothy Bishop, Paolo Struffi, Rimma Levina, Martha L Bulyk, Robert J Johnston, Stephen Small
The K50 (lysine at amino acid position 50) homeodomain (HD) protein Orthodenticle (Otd) is critical for anterior patterning and brain and eye development in most metazoans. In Drosophila melanogaster , another K50HD protein, Bicoid (Bcd), has evolved to replace Otd's ancestral function in embryo patterning. Bcd is distributed as a long-range maternal gradient and activates transcription of a large number of target genes, including otd Otd and Bcd bind similar DNA sequences in vitro, but how their transcriptional activities are integrated to pattern anterior regions of the embryo is unknown...
May 15, 2018: Genes & Development
https://www.readbyqxmd.com/read/29754814/llamatags-a-versatile-tool-to-image-transcription-factor-dynamics-in-live-embryos
#6
Jacques P Bothma, Matthew R Norstad, Simon Alamos, Hernan G Garcia
Embryonic cell fates are defined by transcription factors that are rapidly deployed, yet attempts to visualize these factors in vivo often fail because of slow fluorescent protein maturation. Here, we pioneer a protein tag, LlamaTag, which circumvents this maturation limit by binding mature fluorescent proteins, making it possible to visualize transcription factor concentration dynamics in live embryos. Implementing this approach in the fruit fly Drosophila melanogaster, we discovered stochastic bursts in the concentration of transcription factors that are correlated with bursts in transcription...
April 26, 2018: Cell
https://www.readbyqxmd.com/read/29753017/size-matters-aurora-a-controls-drosophila-larval-development
#7
Lucie Vaufrey, Christine Balducci, René Lafont, Claude Prigent, Stéphanie Le Bras
In metazoans, organisms arising from a fertilized egg, the embryo will develop through multiple series of cell divisions, both symmetric and asymmetric, leading to differentiation. Aurora A is a serine threonine kinase highly involved in such divisions. While intensively studied at the cell biology level, its function in the development of a whole organism has been neglected. Here we investigated the pleiotropic effect of Aurora A loss-of-function in Drosophila larval early development. We report that Aurora A is required for proper larval development timing control through direct and indirect means...
May 9, 2018: Developmental Biology
https://www.readbyqxmd.com/read/29751847/asf1-is-required-to-load-histones-on-the-hira-complex-in-preparation-of-paternal-chromatin-assembly-at-fertilization
#8
Béatrice Horard, Laure Sapey-Triomphe, Emilie Bonnefoy, Benjamin Loppin
BACKGROUND: Anti-Silencing Factor 1 (ASF1) is a conserved H3-H4 histone chaperone involved in both Replication-Coupled and Replication-Independent (RI) nucleosome assembly pathways. At DNA replication forks, ASF1 plays an important role in regulating the supply of H3.1/2 and H4 to the CAF-1 chromatin assembly complex. ASF1 also provides H3.3-H4 dimers to HIRA and DAXX chaperones for RI nucleosome assembly. The early Drosophila embryo is an attractive system to study chromatin assembly in a developmental context...
May 11, 2018: Epigenetics & Chromatin
https://www.readbyqxmd.com/read/29746464/rif1-prolongs-the-embryonic-s-phase-at-the-drosophila-mid-blastula-transition
#9
Charles A Seller, Patrick H O'Farrell
In preparation for dramatic morphogenetic events of gastrulation, rapid embryonic cell cycles slow at the mid-blastula transition (MBT). In Drosophila melanogaster embryos, down-regulation of cyclin-dependent kinase 1 (Cdk1) activity initiates this slowing by delaying replication of heterochromatic satellite sequences and extending S phase. We found that Cdk1 activity inhibited the chromatin association of Rap1 interacting factor 1 (Rif1), a candidate repressor of replication. Furthermore, Rif1 bound selectively to satellite sequences following Cdk1 down-regulation at the MBT...
May 10, 2018: PLoS Biology
https://www.readbyqxmd.com/read/29738712/drosophila-tnf-modulates-tissue-tension-in-the-embryo-to-facilitate-macrophage-invasive-migration
#10
Aparna Ratheesh, Julia Biebl, Jana Vesela, Michael Smutny, Ekaterina Papusheva, S F Gabriel Krens, Walter Kaufmann, Attila Gyoergy, Alessandra Maria Casano, Daria E Siekhaus
Migrating cells penetrate tissue barriers during development, inflammatory responses, and tumor metastasis. We study if migration in vivo in such three-dimensionally confined environments requires changes in the mechanical properties of the surrounding cells using embryonic Drosophila melanogaster hemocytes, also called macrophages, as a model. We find that macrophage invasion into the germband through transient separation of the apposing ectoderm and mesoderm requires cell deformations and reductions in apical tension in the ectoderm...
May 7, 2018: Developmental Cell
https://www.readbyqxmd.com/read/29735716/reducing-resistance-allele-formation-in-crispr-gene-drive
#11
Jackson Champer, Jingxian Liu, Suh Yeon Oh, Riona Reeves, Anisha Luthra, Nathan Oakes, Andrew G Clark, Philipp W Messer
CRISPR homing gene drives can convert heterozygous cells with one copy of the drive allele into homozygotes, thereby enabling super-Mendelian inheritance. Such a mechanism could be used, for example, to rapidly disseminate a genetic payload in a population, promising effective strategies for the control of vector-borne diseases. However, all CRISPR homing gene drives studied in insects thus far have produced significant quantities of resistance alleles that would limit their spread. In this study, we provide an experimental demonstration that multiplexing of guide RNAs can both significantly increase the drive conversion efficiency and reduce germline resistance rates of a CRISPR homing gene drive in Drosophila melanogaster We further show that an autosomal drive can achieve drive conversion in the male germline, with no subsequent formation of resistance alleles in embryos through paternal carryover of Cas9...
May 7, 2018: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/29732657/ocnus-is-essential-for-male-germ-cell-development-in-drosophila-melanogaster
#12
Y Zheng, J Bi, M-Y Hou, W Shen, W Zhang, H Ai, X-Q Yu, Y-F Wang
The ocnus (ocn) gene encodes a protein abundant in the testes, implying its role in testis development. When Drosophila melanogaster is infected with the endosymbiont wMel Wolbachia, which affects the spermatogenesis of its hosts, ocn is downregulated in the third-instar larval testes, suggesting a role of ocn in spermatogenesis. In this study, we knocked down ocn in the testes and found that the hatch rates of embryos derived from ocn-knockdown males were significantly decreased, and 84.38% of the testes were much smaller in comparison to controls...
May 6, 2018: Insect Molecular Biology
https://www.readbyqxmd.com/read/29731302/quantitative-control-of-gpcr-organization-and-signaling-by-endocytosis-in-epithelial-morphogenesis
#13
Ankita Jha, Thomas S van Zanten, Jean-Marc Philippe, Satyajit Mayor, Thomas Lecuit
Tissue morphogenesis arises from controlled cell deformations in response to cellular contractility. During Drosophila gastrulation, apical activation of the actomyosin networks drives apical constriction in the invaginating mesoderm and cell-cell intercalation in the extending ectoderm. Myosin II (MyoII) is activated by cell-surface G protein-coupled receptors (GPCRs), such as Smog and Mist, that activate G proteins, the small GTPase Rho1, and the kinase Rok. Quantitative control over GPCR and Rho1 activation underlies differences in deformation of mesoderm and ectoderm cells...
April 25, 2018: Current Biology: CB
https://www.readbyqxmd.com/read/29727464/patterns-of-chromatin-accessibility-along-the-anterior-posterior-axis-in-the-early-drosophila-embryo
#14
Jenna E Haines, Michael B Eisen
As the Drosophila embryo transitions from the use of maternal RNAs to zygotic transcription, domains of open chromatin, with relatively low nucleosome density and specific histone marks, are established at promoters and enhancers involved in patterned embryonic transcription. However it remains unclear how regions of activity are established during early embryogenesis, and if they are the product of spatially restricted or ubiquitous processes. To shed light on this question, we probed chromatin accessibility across the anterior-posterior axis (A-P) of early Drosophila melanogaster embryos by applying a transposon based assay for chromatin accessibility (ATAC-seq) to anterior and posterior halves of hand-dissected, cellular blastoderm embryos...
May 4, 2018: PLoS Genetics
https://www.readbyqxmd.com/read/29724820/temporal-and-spatial-post-transcriptional-regulation-of-zebrafish-tie1-mrna-by-long-noncoding-rna-during-brain-vascular-assembly
#15
Tamjid A Chowdhury, Chris Koceja, Shahram Eisa-Beygi, Benjamin P Kleinstiver, Suresh N Kumar, Chien-Wei Lin, Keguo Li, Shubhangi Prabhudesai, J Keith Joung, Ramani Ramchandran
OBJECTIVE: Tie1 (tyrosine kinase containing immunoglobulin and epidermal growth factor homology 1), an endothelial and hematopoietic cell-specific receptor tyrosine kinase, is an important regulator of angiogenesis and critical for maintaining vascular integrity. The post-transcriptional regulation of tie1 mRNA expression is not understood, but it might partly explain Tie1's differential expression pattern in endothelium. Following up on our previous work that identified natural antisense transcripts from the tie1 locus- tie1 antisense ( tie1AS ), which regulates tie1 mRNA levels in zebrafish-we attempted to identify the mechanism of this regulation...
May 3, 2018: Arteriosclerosis, Thrombosis, and Vascular Biology
https://www.readbyqxmd.com/read/29724758/evidence-for-the-temporal-regulation-of-insect-segmentation-by-a-conserved-sequence-of-transcription-factors
#16
Erik Clark, Andrew D Peel
Long-germ insects, such as the fruit fly Drosophila melanogaster , pattern their segments simultaneously, whereas short-germ insects, such as the beetle Tribolium castaneum , pattern their segments sequentially, from anterior to posterior. While the two modes of segmentation at first appear quite distinct, much of this difference might simply reflect developmental heterochrony. We now show here that, in both Drosophila and Tribolium , segment patterning occurs within a common framework of sequential Caudal, Dichaete, and Odd-paired expression...
May 3, 2018: Development
https://www.readbyqxmd.com/read/29716356/magnetic-tweezers-optimized-to-exert-high-forces-over-extended-distances-from-the-magnet-in-multicellular-systems
#17
L Selvaggi, L Pasakarnis, D Brunner, C M Aegerter
Magnetic tweezers are mainly divided into two classes depending on the ability of applying torque or forces to the magnetic probe. We focused on the second category and designed a device composed by a single electromagnet equipped with a core having a special asymmetric profile to exert forces as large as 230 pN-2.8 μm Dynabeads at distances in excess of 100 μm from the magnetic tip. Compared to existing solutions our magnetic tweezers overcome important limitations, opening new experimental paths for the study of a wide range of materials in a variety of biophysical research settings...
April 2018: Review of Scientific Instruments
https://www.readbyqxmd.com/read/29715549/selenophosphate-synthetase-1-and-its-role-in-redox-homeostasis-defense-and-proliferation
#18
REVIEW
Jiwoon Na, Jisu Jung, Jeyoung Bang, Qiao Lu, Bradley A Carlson, Xiong Guo, Vadim N Gladyshev, Jinhong Kim, Dolph L Hatfield, Byeong Jae Lee
Selenophosphate synthetase (SEPHS) synthesizes selenophosphate, the active selenium donor, using ATP and selenide as substrates. SEPHS was initially identified and isolated from bacteria and has been characterized in many eukaryotes and archaea. Two SEPHS paralogues, SEPHS1 and SEPHS2, occur in various eukaryotes, while prokaryotes and archaea have only one form of SEPHS. Between the two isoforms in eukaryotes, only SEPHS2 shows catalytic activity during selenophosphate synthesis. Although SEPHS1 does not contain any significant selenophosphate synthesis activity, it has been reported to play an essential role in regulating cellular physiology...
April 28, 2018: Free Radical Biology & Medicine
https://www.readbyqxmd.com/read/29691225/suppression-of-epithelial-folding-at-actomyosin-enriched-compartment-boundaries-downstream-of-wingless-signalling-in-drosophila
#19
Jose M Urbano, Huw W Naylor, Elena Scarpa, Leila Muresan, Bénédicte Sanson
Epithelial folding shapes embryos and tissues during development. Here, we investigate the coupling between epithelial folding and actomyosin-enriched compartmental boundaries. The mechanistic relationship between the two is unclear, because actomyosin-enriched boundaries are not necessarily associated with folds. Also, some cases of epithelial folding occur independently of actomyosin contractility. We investigated the shallow folds called parasegment grooves that form at boundaries between anterior and posterior compartments in the early Drosophila embryo...
April 24, 2018: Development
https://www.readbyqxmd.com/read/29688792/super-resolution-imaging-of-drosophila-tissues-using-expansion-microscopy
#20
Nan Jiang, Hyeon-Jin Kim, Tyler J Chozinski, Jorge E Azpurua, Benjamin A Eaton, Joshua C Vaughan, Jay Z Parrish
The limited resolving power of conventional diffraction-limited microscopy hinders analysis of small, densely packed structural elements in cells. Expansion Microscopy (ExM) provides an elegant solution to this problem, allowing for increased resolution with standard microscopes via physical expansion of the specimen in a swellable polymer hydrogel. Here, we apply, validate, and optimize ExM protocols that enable the study of Drosophila embryos, larval brains, and larval and adult body walls. We achieve a lateral resolution of ∼70 nm in Drosophila tissues using a standard confocal microscope, and we use ExM to analyze fine intracellular structures and intercellular interactions...
April 24, 2018: Molecular Biology of the Cell
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