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Xin Geng, Sandra Acosta, Oleg Lagutin, HyeaJin Gil, Guillermo Oliver
Holoprosencephaly (HPE) is defined as the incomplete separation of the two cerebral hemispheres. The pathology of HPE is variable and based on the severity of the defect, HPE is divided into alobar, semilobar, and lobar. Using a novel hypomorphic Six3 allele, we demonstrate that variability in Six3 dosage results in different HPE phenotypes. Furthermore, we also show that while the semilobar phenotype results from severe downregulation of Shh expression in the rostral diencephalon ventral midline, the alobar phenotype is caused by downregulation of Foxg1 expression in the anterior neural ectoderm...
October 21, 2016: Development
Guangxia Miao, Shigeo Hayashi
Extrinsic branching factors promote the elongation and migration of tubular organs. In the Drosophila tracheal system, Branchless/Fibroblast Growth Factor (FGF) stimulates the branching program by specifying tip cells that acquire motility and lead branch migration to a specific destination. Tip cells have two alternative cell fates: the terminal cell (TC), which elongates the cytoplasmic extension with intracellular lumen, and the fusion cell (FC), which mediates branch connections to form tubular networks...
October 14, 2016: Development
Mara Cucinotta, Silvia Manrique, Andrea Guazzotti, Nadia E Quadrelli, Marta A Mendes, Eva Benkova, Lucia Colombo
The developmental program of the pistil is under the control of both auxin and cytokinin, which crosstalk converges on the regulation of the auxin carrier PIN-FORMED 1 (PIN1). Here we show that in the triple transcription factor mutant cytokinin response factor 2 (crf2) crf3 crf6 both pistil length and ovule number were reduced. PIN1 expression was also lower in the triple mutant and the phenotypes couldn't be rescued by exogenous cytokinin application. pin1 complementation studies using genomic PIN1 constructs showed that the pistil phenotypes were only rescued when the PCRE1 domain, to which CRFs bind, was present...
October 13, 2016: Development
Miquel Sureda-Gómez, José M Martín-Durán, Teresa Adell
The βcatenin-dependent Wnt pathway exerts multiple context-dependent roles in embryonic and adult tissues. In planarians, βcatenin-1 is thought to specify posterior identities through the generation of an anteroposterior gradient. However, the existence of such gradient has not been directly demonstrated. Here, we use a specific polyclonal antibody to demonstrate that nuclear βCATENIN-1 exists as an anteroposterior gradient from the pre-pharyngeal region to the tail of the planarian Schmidtea polychroa High levels in the posterior region steadily decrease towards the pre-pharyngeal region but then increase again in the head region...
October 13, 2016: Development
Hongyuan Shen, Elke Bocksteins, Igor Kondrychyn, Dirk Snyders, Vladimir Korzh
The brain ventricular system is essential for neurogenesis and brain homeostasis. Its neuroepithelial lining effects these functions, but the underlying molecular pathways remain to be understood. We found that the K channels expressed in neuroepithelial cells determine formation of the ventricular system. The phenotype of a novel zebrafish mutant characterized by denudation of neuroepithelial lining of the ventricular system and hydrocephalus is mechanistically linked to Kcng4b, the homologue of the "silent" voltage-gated K channel alpha-subunit Kv6...
October 11, 2016: Development
Richard E Heinz, Michael C Rudolph, Palani Ramanathan, Nicole S Spoelstra, Kiel T Butterfield, Patricia G Webb, Beatrice L Babbs, Hongwei Gao, Shang Chen, Michael A Gordon, Steve M Anderson, Margaret C Neville, Haihua Gu, Jennifer K Richer
Profiling of RNA from mouse mammary epithelial cells (MECs) isolated on pregnancy day 14 (P14) and lactation day 2 (L2) revealed that the majority of differentially expressed microRNA declined precipitously between late pregnancy and lactation. The decline in miR-150, which exhibited the greatest fold decrease, was verified quantitatively and qualitatively. To test the hypothesis that the decline in miR-150 is critical for lactation, MEC-specific constitutive miR-150 was achieved by crossing ROSA26-lox-STOP-lox-miR-150 mice with WAP-driven Cre recombinase mice...
October 11, 2016: Development
T Weber, K Namikawa, B Winter, K Müller-Brown, R Kühn, W Wurst, R W Köster
The zebrafish is a well-established model organism to study in vivo mechanisms of cell communication, differentiation and function. Existing cell ablation methods are either invasive thereby creating additional tissue damage and potential infection sites, or they rely on the cellular expression of prokaryotic enzymes and the use of antibiotic drugs as cell-death-inducing compounds. We have recently established a novel inducible genetic cell ablation system that is based on Tamoxifen-inducible Caspase8-activity, thereby exploiting mechanisms of cell death intrinsic to most cell types...
October 11, 2016: Development
Xing Ma, Yingying Han, Xiaoqing Song, Trieu Do, Zhihao Yang, Jianquan Ni, Ting Xie
Stem cells in adult tissues are constantly exposed to genotoxic stress and also accumulate DNA damage with age. DNA damage has been proposed to cause stem cell loss and cancer formation. However, it remains a mystery how DNA damage leads to both stem cell loss and cancer formation. In this study, we use germline stem cells (GSCs) in the Drosophila ovary to show that DNA damage retards stem cell self-renewal and lineage differentiation in a CHK2 kinase-dependent manner. Both heatshock-inducible endonuclease I-CreI expression and X-ray irradiation can efficiently introduce double-strand breaks in GSCs and their progeny, resulting in a rapid GSC loss and an accumulation of ill-differentiated GSC progeny...
October 11, 2016: Development
Qiang Shao, Stephanie Herrlinger, Si-Lu Yang, Fan Lai, Julie M Moore, Melinda A Brindley, Jian-Fu Chen
Zika virus (ZIKV) infection of pregnant women can result in fetal brain abnormalities. It has been established that ZIKV disrupts neural progenitor cells (NPCs) and leads to embryonic microcephaly. However, the fate of other cell types in the developing brain and their contributions to ZIKV-associated brain abnormalities remain largely unknown. Using intracerebral inoculation of embryonic mouse brains, we found that ZIKV infection leads to postnatal growth restriction including microcephaly. In addition to cell cycle arrest and apoptosis of NPCs, ZIKV infection causes massive neuronal death and axonal rarefaction which phenocopy fetal brain abnormalities in humans...
October 11, 2016: Development
Emily Swanzey, Matthias Stadtfeld
Genomic imprinting results in the monoallelic expression of genes that encode important regulators of growth and proliferation. Dysregulation of imprinted genes, such as those within the Dlk1-Dio3 locus, is associated with developmental syndromes and specific diseases. Our ability to interrogate causes of imprinting instability has been hindered by the absence of suitable model systems. Here, we describe a Dlk1 knockin reporter mouse that enables single-cell visualization of allele-specific expression and prospective isolation of cells, simultaneously...
October 11, 2016: Development
Dolores Irala, Antonela Bonafina, Paula Aldana Fontanet, Fernando Cruz Alsina, Gustavo Paratcha, Fernanda Ledda
The formation of synaptic connections during nervous system development requires the precise control of dendrite growth and synapse formation. Although Glial Cell line-Derived Neurotrophic Factor (GDNF) and its receptor GFRα1 are expressed in the forebrain, the role of this system in hippocampus remains unclear. In this work, we investigated the consequences of GFRα1 deficiency for the development of hippocampal connections. Analysis of conditional GFRα1-knockout mice shows a reduction in dendritic length and complexity, as well as a decrease in postsynaptic density (PSD) specializations and in the synaptic localization of postsynaptic proteins in hippocampal neurons...
October 5, 2016: Development
Kaoru S Imai, Clare Hudson, Izumi Oda-Ishii, Hitoyoshi Yasuo, Yutaka Satou
Many animal embryos use nuclear β-catenin (nβ-catenin) during the segregation of endomesoderm (or endoderm) from ectoderm. This mechanism is thus likely to be evolutionarily ancient. In the ascidian embryo, nβ-catenin reiteratively drives binary fate decisions between ectoderm and endomesoderm at the 16-cell stage, and then between endoderm and margin (mesoderm and caudal neural) at the 32-cell stage. At the 16-cell stage, nβ-catenin activates endomesoderm genes in the vegetal hemisphere. At the same time, nβ-catenin suppresses the DNA-binding activity of a maternal transcription factor Gata...
October 5, 2016: Development
Jose Arturo Gutierrez-Triana, Juan L Mateo, David Ibberson, Soojin Ryu, Joachim Wittbrodt
DNA adenine methyltransferase identification (DamID) has emerged as an alternative method to profile protein-DNA interactions; however, critical issues limit its widespread applicability. Here we present iDamIDseq, a protocol that improves specificity and sensitivity by inverting the steps DpnI-DpnII and adding steps involving a phosphatase and exonuclease. To determine genome-wide protein-DNA interactions efficiently, we present the analysis tool iDEAR (iDamIDseq Enrichment Analysis with R). The combination of DamID and iDEAR permits the establishment of consistent profiles for transcription factors, even in transient assays, as we exemplify using the small teleost medaka (Oryzias latipes)...
October 5, 2016: Development
Anna Melkov, Raju Baskar, Yehonatan Alcalay, Uri Abdu
Intrinsic cell microtubule (MT) polarity, together with molecular motors and adaptor proteins, determines mitochondrial polarized targeting and MT-dependent transport. In polarized cells, such as neurons, mitochondrial mobility and transport require the regulation of kinesin and dynein by two adaptor proteins, Milton and Miro. Recently, we found that dynein heavy chain 64C (Dhc64C) is the primary motor protein for both anterograde and retrograde transport of mitochondria in the Drosophila bristle. In this study, we revealed that a molecular lesion in the Dhc64C allele that reduced bristle mitochondrial velocity generated a variant that acts as a "slow" dynein in a MT gliding assay, indicative of dynein directly regulating mitochondrial transport...
October 5, 2016: Development
Alys M Cheatle Jarvela, Kristen A Yankura, Veronica F Hinman
How neural stem cells generate the correct number and type of differentiated neurons in appropriate places is an important question in developmental biology. Although nervous systems are diverse across phyla, many taxa have a larva that forms an anterior concentration of serotonergic neurons, or apical organ. The number of neurons in these organs is highly variable. Previous work demonstrated that the sea star embryo initially has a pan-neurogenic ectoderm, but the genetic mechanism that directs only a subset of these cells to generate serotonergic neurons in a particular location had not been resolved...
October 5, 2016: Development
Amir Mizbani, Edlira Luca, Elisabeth J Rushing, Jan Krützfeldt
MicroRNAs (miRNAs) are important regulators of skeletal muscle regeneration, but the underlying mechanisms are still incompletely understood. Here, comparative miRNA sequencing analysis of myogenic progenitor cells (MPs) and non-myogenic fibroblast-adipocyte progenitors (FAPs) during cardiotoxin (CTX)-induced muscle injury uncovered miR-501 as a novel muscle-specific miRNA. miR-501 is an intronic miRNA and its expression levels in MPs correlated with its host gene, chloride channel, voltage-sensitive 5, Clcn5 Pharmacological inhibition of miR-501 dramatically blunted the induction of embryonic myosin heavy chain (MYH3) and, to a lesser extent, adult myosin isoforms during muscle regeneration and promoted small-diameter neofibers...
October 5, 2016: Development
Daniel D Shaye, Iva Greenwald
The C. elegans excretory cell (EC) is a powerful model for tubulogenesis, a conserved process that requires precise cytoskeletal regulation. EXC-6, an ortholog of the disease-associated formin INF2, coordinates cell outgrowth and lumen formation during EC tubulogenesis by regulating F-actin at the tip of the growing canal and the dynamics of basolateral microtubules. EXC-6 functions in parallel to EXC-5/FGD, a predicted activator of the Rho GTPase Cdc42. Here we identify the parallel pathway: EXC-5 functions through CDC-42 to regulate two other formins: INFT-2, another INF2 ortholog, and CYK-1, the sole ortholog of mammalian Diaphanous (mDia)...
October 3, 2016: Development
Courtney R Bone, Yu-Tai Chang, Natalie E Cain, Shaun P Murphy, Daniel A Starr
Cellular migrations through constricted spaces are a critical aspect of many developmental and disease processes including hematopoiesis, inflammation, and metastasis. A limiting factor in these events is nuclear deformation. Here, we establish an in vivo model where nuclei can be visualized while moving through constrictions and use it to elucidate mechanisms for nuclear migration. C. elegans hypodermal P-cell larval nuclei traverse a narrow space about 5% their width. This constriction is blocked by fibrous organelles, structures connecting the muscles to cuticle through P cells...
October 3, 2016: Development
Michaela Matthes, Ramón A Torres-Ruiz
Several observations suggest that the micronutrient boron (B) has a stabilizing role in the plasma membrane (PM), supporting functions in PM-linked (hormone) signalling processes. However, this role is poorly characterized. This study shows, that boronic acids, specific competitors of B, induce phenocopies of the Arabidopsis thaliana rootless pattern mutant monopteros At least in part, this is caused by phenylboronic acid (PBA)-induced internalisation of the membrane-localized auxin efflux carrier PINFORMED1 (PIN1) in the early embryo...
October 3, 2016: Development
Keren Gueta, Ahuvit David, Tsadok Cohen, Yotam Menuchin-Lasowski, Hila Nobel, Ginat Narkis, LiQi Li, Paul Love, Jimmy de Melo, Seth Blackshaw, Heiner Westphal, Ruth Ashery-Padan
The Lim domain binding proteins (Ldbs) are key cofactor proteins that assemble with LIM domains of the LMO/LIM-HD family to form functional complexes that regulate cell proliferation and differentiation throughout the CNS. Here, using conditional mutagenesis and comparative phenotypic analysis, we analyze the function of Ldb1 and Ldb2 in mouse retinal development, and demonstrate overlapping and specific functions of both genes. We show that Ldb1 interacts with Lhx2 in the embryonic retina and that both Ldb1 and Ldb2, probably functioning with Lhx2 in a complex, play a key role in maintaining the pool of retinal progenitor cells...
October 3, 2016: Development
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