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April R Reedy, Liping Luo, Andrew S Neish, Rheinallt M Jones
A distinct taxon of the Drosophila microbiota, Lactobacillus plantarum , is capable of stimulating the generation of reactive oxygen species (ROS) within cells, and inducing epithelial cell proliferation. Here we show microbial-induced ROS generation within Drosophila larval stem cell compartments exhibits a distinct spatial distribution. Lactobacilli-induced ROS is strictly excluded from defined midgut compartments that harbor adult midgut progenitor (AMP) cells, forming a functional "ROS sheltered zone" (RSZ)...
January 18, 2019: Development
Boris Reznik, Steven A Cincotta, Rebecca G Jaszczak, Leslie J Mateo, Joel Shen, Mei Cao, Laurence Baskin, Ping Ye, Wenfeng An, Diana J Laird
Epigenetic resetting in germ cells during development de-represses transposable elements (TEs). piRNAs protect fetal germ cells by targeted mRNA destruction and deposition of repressive epigenetic marks. Here we provide the first evidence for an active piRNA pathway and TE repression in germ cells of human fetal testis. We identify pre-pachytene piRNAs with features of secondary amplification that map most abundantly to the long interspersed element type 1 (L1) family of TEs. L1-ORF1p expression is heterogeneous in fetal germ cells, peaks at mid-gestation and declines concomitantly with increases in piRNAs, nuclear localization of HIWI2, and H3K9me3...
January 18, 2019: Development
Elena M Popa, Marcela Buchtova, Abigail S Tucker
Most mammals have two sets of teeth (diphyodont), a deciduous dentition replaced by a permanent dentition, however the mouse possesses only one tooth generation (monophyodont). In diphyodonts the replacement tooth forms on the lingual side of the first tooth from the successional dental lamina. This lamina expresses the stem/progenitor marker Sox2 and has activated Wnt/B-catenin signalling at its tip. Although the mouse does not replace its teeth a transient rudimentary successional dental lamina (RSDL) still forms during development...
January 18, 2019: Development
Dimitrios K Papadopoulos, Kassiani Skouloudaki, Ylva Engström, Lars Terenius, Rudolf Rigler, Christoph Zechner, Vladana Vukojević, Pavel Tomancak
The variability in transcription factor concentration among cells is an important developmental determinant, yet how variability is controlled remains poorly understood. Studies of variability have focused predominantly on monitoring mRNA production noise. Little information exists about transcription factor protein variability, since this requires the use of quantitative methods with single-molecule sensitivity. Using Fluorescence Correlation Spectroscopy (FCS), we characterized the concentration and variability of 14 endogenously tagged TFs in live Drosophila imaginal discs...
January 14, 2019: Development
Noelia Muñoz-Martín, Rocío Sierra, Thomas Schimmang, Cristina Villa Del Campo, Miguel Torres
Myc is considered an essential transcription factor for heart development, but cardiac defects have only been studied in global Myc loss of function models. Here, we eliminated Myc by recombining a Myc floxed allele with the Nkx2.5Cre driver. We observed no anatomical, cellular or functional alterations in either fetuses or adult cardiac Myc-deficient mice. We re-examined Myc expression during development and found no expression in developing cardiomyocytes. In contrast, we confirmed that Mycn is essential for cardiomyocyte proliferation and cardiogenesis...
January 14, 2019: Development
Tabea Schilling, Aicha H Ali, Aljoscha Leonhardt, Alexander Borst, Jesús Pujol-Martí
In the Drosophila visual system, T4/T5 neurons represent the first stage in which the direction of visual motion is computed. T4 and T5 neurons exist in four subtypes, each responding to motion in one of the four cardinal directions and projecting axons into one of the four lobula plate layers. However, all T4/T5 neurons share properties essential for sensing motion. How T4/T5 neurons acquire their properties during development is poorly understood. We reveal that SoxN and Sox102F transcription factors control the acquisition of properties common to all T4/T5 neuron subtypes, i...
January 14, 2019: Development
Koji Ando, Weili Wang, Di Peng, Ayano Chiba, Anne Lagendijk, Lindsey Barske, J Gage Crump, Didier Y R Stainier, Urban Lendahl, Kaska Koltowska, Benjamin M Hogan, Shigetomo Fukuhara, Naoki Mochizuki, Christer Betsholtz
Mural cells (MCs) are essential for blood vessel stability and function; however, the mechanisms regulating MC development remain incompletely understood, particularly those involved in MC specification. Here, we investigated the first steps of MC formation in zebrafish utilizing transgenic reporters. Using pdgfrb and abcc9 reporters, we show that the onset of expression of abcc9 , a pericyte marker in adult mice and zebrafish, occurs almost coincidentally with an increment in pdgfrb expression in peri-arterial mesenchymal cells , suggesting that these transcriptional changes mark the specification of MC lineage cells from naïve pdgfrb low mesenchymal cells...
January 14, 2019: Development
Rui Yang, Huanmin Wang, Boxi Kang, Bin Chen, Yaoyao Shi, Shuchun Yang, Lihong Sun, Yufang Liu, Weidi Xiao, Tao Zhang, Juntao Yang, Ye Zhang, Mingzhao Zhu, Ping Xu, Yongsheng Chang, Yuyan Jia, Yue Huang
Protein modification by ubiquitin and ubiquitin-like proteins (UBLs) regulates numerous biological functions. The UFM1 system, a novel UBL conjugation system, is implicated in mouse development and hematopoiesis. However, its broad biological functions and working mechanisms remain largely elusive. CDK5RAP3, a possible ufmylation substrate, is essential for epiboly and gastrulation in zebrafish. Herein, we report a critical role of CDK5RAP3 in liver development and hepatic functions. Cdk5rap3 knockout mice displayed prenatal lethality with severe liver hypoplasia, as characterized by delayed proliferation and compromised differentiation...
January 11, 2019: Development
Weronika Fic, Celia Faria, Daniel St Johnston
The timing of Drosophila egg chamber development is controlled by a germline Delta signal that activates Notch in the follicle cells to induce them to cease proliferation and differentiate. Here we report that follicle cells lacking the RNA-binding protein IMP go through one extra division due to a delay in the Delta-dependent S2 cleavage of Notch. The timing of Notch activation has previously been shown to be controlled by cis-inhibition by Delta in the follicle cells, which is relieved when the miRNA pathway represses Delta expression...
January 11, 2019: Development
Matthew Hannaford, Nicolas Loyer, Francesca Tonelli, Martin Zoltner, Jens Januschke
Studying the function of proteins using genetics in cycling cells is complicated by the fact that there is often a delay between gene inactivation and the timepoint of phenotypic analysis. This is particularly true when studying kinases, that have pleiotropic functions and multiple substrates. Drosophila neuroblasts are rapidly dividing stem cells and an important model system to study cell polarity. Mutations in multiple kinases cause neuroblast polarity defects, but their precise functions at particular time points in the cell cycle are unknown...
January 11, 2019: Development
Harinarayana Ankamreddy, Hyehyun Min, Jae Yoon Kim, Xiao Yang, Eui-Sic Cho, Un-Kyung Kim, Jinwoong Bok
Defects in the middle ear ossicles - malleus, incus, and stapes - can lead to conductive hearing loss. During development, neural crest cells (NCCs) migrate from the dorsal hindbrain to specific locations in pharyngeal arch (PA) 1 and 2, to form the malleus-incus and stapes, respectively. It is unclear how migratory NCCs reach their proper destination in PA and initiate mesenchymal condensation to form specific ossicles. We show that secreted molecules sonic hedgehog (SHH) and bone morphogenetic protein 4 (BMP4) emanating from the pharyngeal endoderm are important in instructing regional-specific NCC condensation to form malleus-incus and stapes, respectively...
January 10, 2019: Development
Masanori Kawaguchi, Kota Sugiyama, Kazumi Matsubara, Che-Yi Lin, Shigehiro Kuraku, Shota Hashimoto, Yoshiaki Suwa, Luok Wen Yong, Koji Takino, Shota Higashida, Daisuke Kawamura, Jr-Kai Yu, Yoshiyuki Seki
Gene regulatory networks underlying cellular pluripotency are controlled by a core circuitry of transcription factors in mammals, including POU5F1. However, the evolutionary origin and transformation of pluripotency-related transcriptional networks have not been elucidated in deuterostomes. PR domain-containing protein 14 (PRDM14) is specifically expressed in pluripotent cells and germ cells, and required for establishing embryonic stem cells (ESCs) and primordial germ cells in mice. Here, we compared the functions and expression patterns of PRDM14 orthologues within deuterostomes...
January 10, 2019: Development
Florian Geisler, Richard A Coch, Christine Richardson, Martin Goldberg, Bernd Denecke, Olaf Bossinger, Rudolf E Leube
The enrichment of intermediate filaments in the apical cytoplasm of intestinal cells is evolutionary conserved forming a sheath that is anchored to apical junctions and positioned below the microvillar brush border suggestive of a protective intracellular barrier function. To test this, we used C. elegans , whose intestinal cells are endowed with a particularly dense intermediate filament-rich layer that is referred to as the endotube. We find alterations in endotube structure and intermediate filament expression upon infection with nematicidal Bacillus thuringiensis or treatment with its major pore-forming toxin crystal protein Cry5B...
January 10, 2019: Development
Yiyun Zhou, Sarah E Popadowski, Emily Deustchman, Marc S Halfon
Pleiotropic signaling pathways must somehow engender specific cellular responses. In the Drosophila mesoderm, Ras pathway signaling specifies muscle founder cells from among the broader population of myoblasts. For somatic muscles, this is an inductive process mediated by the ETS-domain downstream Ras effectors Pointed and Aop (Yan). We demonstrate here that for the circular visceral muscles, despite superficial similarities, a significantly different specification mechanism is at work. Not only is visceral founder cell specification not dependent on Pointed or Aop, but Ras pathway signaling in its entirety can be bypassed...
January 10, 2019: Development
Ana Luisa Rodrigues Toste de Carvalho, Alexandros Strikoudis, Hsiao-Yun Liu, Ya-Wen Chen, Tiago J Dantas, Richard B Vallee, Jorge Correia-Pinto, Hans-Willem Snoeck
Although strategies for directed differentiation of human pluripotent stem cells (hPSCs) into lung and airway have been established, terminal maturation of the cells remains a vexing problem. We show here that in Collagen I 3D cultures in the absence of glycogen synthase kinase 3 (GSK3) inhibition, hPSC-derived lung progenitors (LPs) undergo multilineage maturation into proximal cells, type I alveolar epithelial cells and morphologically mature type II cells. Enhanced cell cycling, one of the signaling outputs of GSK3 inhibition, plays a role in the maturation-inhibiting effect of GSK3 inhibition...
December 21, 2018: Development
Elizabeth E Capowski, Kayvan Samimi, Steven J Mayerl, M Joseph Phillips, Isabel Pinilla, Sara E Howden, Jishnu Saha, Alex D Jansen, Kimberly L Edwards, Lindsey D Jager, Katherine Barlow, Rasa Valiauga, Zachary Erlichman, Anna Hagstrom, Divya Sinha, Valentin M Sluch, Xitiz Chamling, Donald J Zack, Melissa C Skala, David M Gamm
Numerous protocols have been described that produce neural retina from human pluripotent stem cells (hPSCs), many of which are based on the culture of 3D organoids. While nearly all such methods yield at least partial segments of highly mature-appearing retinal structure, variabilities exist within and between organoids that can change over a protracted time course of differentiation. Adding to this complexity are potential differences in the composition and configuration of retinal organoids when viewed across multiple differentiations and hPSC lines...
December 19, 2018: Development
Georg Wolfstetter, Ina Dahlitz, Kathrin Pfeifer, Uwe Töpfer, Joscha Arne Alt, Daniel Christoph Pfeifer, Reinhard Lakes-Harlan, Stefan Baumgartner, Ruth H Palmer, Anne Holz
Basement membranes (BMs) are specialized layers of extracellular matrix (ECM) mainly composed of Laminin, type IV Collagen, Perlecan and Nidogen/entactin (NDG). Recent in vivo studies challenged the initially proposed role of NDG as major ECM linker molecule by revealing dispensability for viability and BM formation. Here, we report the characterization of the single Ndg gene in Drosophila. Embryonic Ndg expression was primarily observed in mesodermal tissues and the chordotonal organs, whereas NDG protein localized to all BMs...
December 19, 2018: Development
Filip J Wymeersch, Stavroula Skylaki, Yali Huang, Julia A Watson, Constantinos Economou, Carylyn Marek-Johnston, Simon R Tomlinson, Valerie Wilson
The elongating mouse anteroposterior axis is supplied by progenitors with distinct tissue fates. It is not known whether these progenitors confer anteroposterior pattern to the embryo. We have analysed the progenitor population transcriptomes in the mouse primitive streak and tail bud throughout axial elongation. Transcriptomic signatures distinguish three known progenitor types (neuromesodermal, lateral/paraxial mesoderm and notochord progenitors; NMPs, LPMPs and NotoPs). Both NMP and LPMP transcriptomes change extensively over time...
December 17, 2018: Development
Andrew M Hudson, Katelynn M Mannix, Julianne A Gerdes, Molly C Kottemann, Lynn Cooley
During Drosophila oogenesis, specialized actin-based structures called ring canals form and expand to accommodate growth of the oocyte. Previous work demonstrated that Kelch and Cullin 3 function together in a Cullin 3-RING ubiquitin ligase complex (CRL3Kelch ) to organize the ring canal cytoskeleton, presumably by targeting a substrate for proteolysis. Here, we use tandem affinity purification followed by mass spectrometry to identify HtsRC as the CRL3Kelch ring canal substrate. CRISPR-mediated mutagenesis of HtsRC revealed its requirement in the recruitment of the ring canal F-actin cytoskeleton...
December 17, 2018: Development
Anna F Gilles, Johannes B Schinko, Magdalena I Schacht, Camille Enjolras, Michalis Averof
Clonal marking techniques based on the Cre/lox and Flp/FRT systems are widely used in multicellular model organisms to mark individual cells and their progeny, in order to study their morphology, growth properties and developmental fates. The same tools can be adapted to introduce specific genetic changes in a subset of cells within the body, i.e. to perform mosaic genetic analysis. Marking and manipulating distinct cell clones requires control over the frequency of clone induction, which is sometimes difficult to achieve...
December 14, 2018: Development
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