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Genesis: the Journal of Genetics and Development

Emmanuel Villanueva-Chimal, Laura S Salinas, Laura P Fernández-Cardenas, Gabriela Huelgas-Morales, Alejandro Cabrera-Wrooman, Rosa E Navarro
The d4 family of transcription factors consists of three members in mammals. DPF1/neuro-d4 is expressed mainly in neurons and the peripheral nervous system, and is important for brain development. DPF2/requiem/ubi-d4 is expressed ubiquitously and presumably functions as an apoptotic factor, especially during the deprivation of trophic factors. DPF3/cer-d4 is expressed in neurons and in the heart, and is important for heart development and function in zebrafish. In Drosophila, there is only one member, dd4, whose function is still unknown, but it is expressed in many tissues and is particularly abundant in the brain of developing embryos and in adults...
September 23, 2017: Genesis: the Journal of Genetics and Development
Jordan J Toutounchian, Joseph H McCarty
Perivascular astrocyte end feet closely juxtapose cerebral blood vessels to regulate important developmental and physiological processes including endothelial cell proliferation and sprouting as well as the formation of the blood-brain barrier (BBB). The mechanisms underlying these events remain largely unknown due to a lack of experimental models for identifying perivascular astrocytes and distinguishing these cell types from other astroglial populations. Megalencephalic leukoencephalopathy with subcortical cysts 1 (Mlc1) is a transmembrane protein that is expressed in perivascular astrocyte end feet where it controls BBB development and homeostasis...
September 20, 2017: Genesis: the Journal of Genetics and Development
Harunur Rashid, Haiyan Chen, Mohammad Hassan, Amjad Javed
Aggrecan (Acan), a large proteoglycan is abundantly expressed in cartilage tissue. Disruption of Acan gene causes dwarfism and perinatal lethality of homozygous mice. Due to sustained expression of Acan in the growth plate and articular cartilage, Agc(Cre) model has been developed for the regulated ablation of target gene in chondrocytes. In this model, the IRES-CreERT-Neo-pgk transgene is knocked-in the 3'UTR of the Acan gene. We consistently noticed variable weight and size among the Agc(Cre) littermates, prompting us to examine the cause of this phenotype...
September 18, 2017: Genesis: the Journal of Genetics and Development
Ana Paço, Renata Freitas
Genes in the 5' extremity of the HoxD cluster encode DNA-binding transcription factors essential for development of the autopod and digits, regulating primarily gene expression and, consequently, morphogenesis and skeletal differentiation. Comparative studies focused on their expression and regulation have led to the idea that evolution of a bimodal regulation of the HoxD cluster, mainly due to the activation of cis-regulatory units in the centromeric side of the cluster, was a fundamental mechanism that potentiated the fin-to-limb transition in vertebrates...
September 14, 2017: Genesis: the Journal of Genetics and Development
Nicholas W Plummer, Erica K Ungewitter, Kathleen G Smith, Humphrey H-C Yao, Patricia Jensen
Recombinase responsive mouse lines expressing diphtheria toxin subunit A (DTA) are well established tools for targeted ablation of genetically defined cell populations. Here we describe a new knock-in allele at the Gt(Rosa)26Sor locus that retains the best features of previously described DTA alleles-including a CAG promoter, attenuated mutant DTA cDNA, and ubiquitous EGFP labeling-with the addition of a Cre-dependent FLEx switch for tight control of expression. The FLEx switch consists of two pairs of antiparallel lox sites requiring Cre-mediated recombination for inversion of the DTA to the proper orientation for transcription...
September 5, 2017: Genesis: the Journal of Genetics and Development
Jolanda J D de Roo, Cor Breukel, Amiet R Chhatta, Margot M Linssen, Sandra A Vloemans, Daniela Salvatori, Harald M M Mikkers, Sjef J Verbeek, Frank J T Staal
The canonical Wnt signalling pathway has been implicated in organogenesis and self-renewal of essentially all stem cell systems. In vivo reporter systems are crucial to assess the role of Wnt signalling in the biology and pathology of stem cell systems. We set out to develop a Turquoise (TQ) fluorescent protein based Wnt reporter. We used a CRISPR-Cas9 approach to insert a turquoise fluorescent protein encoding gene into the general Wnt target gene Axin2, thereby establishing a Wnt reporter mouse similar to previously generated Wnt reporter mice but with the mTurquoise2 gene instead of E...
September 5, 2017: Genesis: the Journal of Genetics and Development
Mingmei Liao, Junmei Zhou, Fen Wang, Yasmin H Ali, Kelvin L Chan, Fei Zou, Stefan Offermanns, Zhisheng Jiang, Zhihua Jiang
The Myh11-CreER(T2) mouse line (Cre(+) ) has gained increasing application due to its high lineage specificity relative to other Cre drivers targeting smooth muscle cells (SMCs). This Cre allele, however, was initially inserted into the Y chromosome (X/Y(Cre+) ), which excluded its application in female mice. Our group established a Cre(+) colony from male ancestors. Surprisingly, genotype screening identified female carriers that stably transmitted the Cre allele to the following generations. Crossbreeding experiments revealed a pattern of X-linked inheritance for the transgene (k>1000), indicating that these female carries acquired the Cre allele through a mechanism of Y to X chromosome translocation...
August 28, 2017: Genesis: the Journal of Genetics and Development
Yacine Kherdjemil, Marie Kmita
Tetrapods are characterized by the presence of digits at the distal end of their limbs, which have emerged during the transition from fins to limbs. While variations in digit number are observed in extant tetrapods, most have five digits per limb and divergence from this pentadactyl ground state is always a reduction in digit number. Paleontological data revealed that stem-group tetrapods were polydactylous indicating that the evolution from fish fin to modern tetrapod limbs involved two major transitions; the emergence of digits and the shift from polydactyly to pentadactyly...
August 24, 2017: Genesis: the Journal of Genetics and Development
Danielson B Amaral, Igor Schneider
Limbs with digits evolved as sarcopterygian fish transitioned to a terrestrial life, giving rise to modern tetrapods. Since the Devonian, most of the sarcopterygian fish diversity became extinct, with the only surviving representatives being two coelacanth and six lungfish species. As the sister group of tetrapods, sarcopterygian fish constitute the ideal models to address questions regarding the transition of vertebrates from water to land. However, distantly-related yet experimentally-amenable teleost fish species have instead become the organisms of choice for geneticists and developmental biologists...
August 19, 2017: Genesis: the Journal of Genetics and Development
Mikiko Tanaka
The evolution from fins to limbs was one of the most successful innovations for vertebrates, allowing them to vastly expand their behaviors and habitats. Fossil records suggest that morphological changes occurred not only along the proximal-distal axis included appearance of the autopod, but also occurred along the anterior-posterior axis included reductions in the size and number of basal bones and digits. This review focuses on recent progress in developmental and genetic studies aimed at elucidating the mechanisms underlying alteration of anterior-posterior patterning and its accompanying changes along the proximal-distal axis during the fin-to-limb transition...
August 19, 2017: Genesis: the Journal of Genetics and Development
Fuki Gyoja
Basic helix-loop-helix (bHLH) transcription factors have attracted the attention of developmental and evolutionary biologists for decades because of their conserved functions in mesodermal and neural tissue formation in both vertebrates and fruit flies. Their evolutionary history is of special interest because it will likely provide insights into developmental processes and refinement of metazoan-specific traits. This review briefly considers advances in developmental biological studies on bHLHs/HLHs. I also discuss recent genome-wide surveys and molecular phylogenetic analyses of these factors in a wide range of metazoans...
August 14, 2017: Genesis: the Journal of Genetics and Development
Matthew E Bechard, Eric D Bankaitis, Alessandro Ustione, David W Piston, Mark A Magnuson, Christopher V E Wright
During pancreas organogenesis, Neurog3(HI) endocrine-committing cells are generated from a population of Sox9(+) mitotic progenitors with only a low level of Neurog3 transcriptional activity (Neurog3(TA.LO) ). Low-level Neurog3 protein, in Neurog3(TA.LO) cells, is required to maintain their mitotic endocrine-lineage-primed status. Herein, we describe a Neurog3-driven FUCCI cell-cycle reporter (Neurog3(P2A.FUCCI) ) derived from a Neurog3 BAC transgenic reporter that functions as a loxed cassette acceptor (LCA)...
August 2, 2017: Genesis: the Journal of Genetics and Development
Matthew Towers
The developing limbs of chicken embryos have served as pioneering models for understanding pattern formation for over a century. The ease with which chick wing and leg buds can be experimentally manipulated, while the embryo is still in the egg, has resulted in the discovery of important developmental organisers, and subsequently, the signals that they produce. Sonic hedgehog (Shh) is produced by mesenchyme cells of the polarizing region at the posterior margin of the limb bud and specifies positional values across the antero-posterior axis (the axis running from the thumb to the little finger)...
July 22, 2017: Genesis: the Journal of Genetics and Development
Bipin K Tripathi, Rituparna Das, Ashim Mukherjee, Mousumi Mutsuddi
Identification of Spoon as a suppressor of SCA8 associated neurodegeneration provides us a hint about its role in neuronal development and maintenance. However, a detailed molecular characterization of spoon has not yet been reported. Here, we describe spatial expression pattern of Spoon during Drosophila development. Quantitative real time-PCR and fluorescent RNA-RNA in situ hybridization indicate that Spoon is expressed at relatively high levels in larval brain and photoreceptors of eye-antennal discs. Immunostaining reveals that Spoon is subcellularly localized in the cytoplasm and is also membrane bound...
July 19, 2017: Genesis: the Journal of Genetics and Development
Yun He, Xiumei Sun, Li Wang, Yuji Mishina, Jun-Lin Guan, Fei Liu
Conditional gene knockout using the Cre/loxP system is instrumental in advancing our understanding of the function of genes in a wide range of disciplines. It is becoming increasingly apparent in the literature that recombination mediated by some Cre transgenes can occur in unexpected tissues. Dermo1-Cre (Twist2-Cre) has been widely used to target skeletal lineage cells as well as other mesoderm-derived cells. Here we report that Dermo1-Cre exhibits spontaneous male germline recombination activity leading to a Cre-mediated recombination of a floxed Ptk2 (Protein tyrosine kinase 2, also known as Fak [Focal adhesion kinase]) allele but not a floxed Rb1cc1 (RB1 inducible coiled-coil 1, also known as Fip200 [FAK-family Interacting Protein of 200 kDa]) allele at high frequency...
September 2017: Genesis: the Journal of Genetics and Development
Praween Kumar Choubey, Jagat Kumar Roy
Intracellular vesicular trafficking is one of the important tools in maintaining polarity, adhesion, and shape of epithelial cells. Rab11, a subfamily of the Ypt/Rab gene family of ubiquitously expressed GTPases and a molecular marker of recycling endosomes, transports different components of plasma membrane. Here, we report that Rab11 affects tubulogenesis of Malpighian tubules (MTs). MTs are simple polarized epithelial tubular structures, considered as functional analogue of human kidney. Rab11 has pleiotropic effects on MTs development as down-regulation of Rab11 in principal cells (PCs) of MTs from embryonic stages of development results in reduced endoreplication, clustering of cells, disorganized cytoskeleton, and disruption of polarity leading to shortening of MTs in third instar larvae...
August 2017: Genesis: the Journal of Genetics and Development
Thomas P Clements, Bhavna Tandon, Hendrik A Lintel, Joseph H McCarty, Daniel S Wagner
Application of CRISPR-Cas9 technology in diverse organisms has resulted in an explosion of genome modification efforts. To expand the toolbox of applications, we have created an E. coli Exonuclease I (sbcB)-Cas9 fusion that has altered enzymatic activity in zebrafish embryos. This Cas9 variant has increased mutation efficiency and favors longer deletions relative to wild-type Cas9. We anticipate that this variant will allow for more efficient screening for F0 phenotypes and mutation of a larger spectrum of genomic targets including deletion of regulatory regions and creating loss of function mutations in transcription units with poor sequence conservation such as lncRNAs where larger deletions may be required for loss of function...
August 2017: Genesis: the Journal of Genetics and Development
Simone Probst, Ray A Daza, Natalie Bader, Jonas F Hummel, Matthias Weiß, Yakup Tanriver, Robert F Hevner, Sebastian J Arnold
The T-box transcription factor Eomes (also known as Tbr2) shows short-lived expression in various localized domains of the embryo, including epiblast cells during gastrulation and intermediate progenitor cells in the cerebral cortex. In these tissues Eomes fulfills crucial roles for lineage specification of progenitors. To directly observe Eomes-dependent cell lineages in the living embryo, we generated a novel dual-fluorescence reporter allele that expresses a membrane-bound tdTomato protein for investigation of cell morphology and a nuclear GFP for cell tracing...
August 2017: Genesis: the Journal of Genetics and Development
Noriko Hamada-Kawaguchi, Daisuke Yamamoto
Drosophila Btk29A is a Tec family nonreceptor tyrosine kinase, the ortholog of which causes X-linked agammaglobulinemia in humans when mutant. In Btk29A(ficP) mutant ovaries, multiple defects are observed: extrapolar cells form ectopically; osk mRNA fails to accumulate posteriorly in mature oocytes; the shape and alignment of follicle cells are grossly distorted. All these phenotypes are rescued by selectively overexpressing the type 2 isoform of wild-type Btk29A in follicle cells. Expression of certain proteins enriched in adherens junctions is markedly affected in Btk29A(ficP) mutants; the anterior-posterior gradient normally observed in the expression of DE-Cadherin and Armadillo are lost and Canoe is sequestered from adherens junctions...
August 2017: Genesis: the Journal of Genetics and Development
Sara Ranjbarvaziri, Shuin Park, Ngoc B Nguyen, William B Gilmore, Peng Zhao, Reza Ardehali
Nkx2-5 is a homeobox-containing transcriptional regulator that serves as one of the earliest markers of cardiac lineage commitment. To study the role of Nkx2-5-expressing progenitors at specific time points in cardiac development, we have generated a novel and inducible NKX2-5 mouse line by knocking in a CreER cassette into the Nkx2-5 genomic locus, while preserving the endogenous Nkx2-5 gene to avoid haploinsufficiency. We evaluated the specificity and efficiency of CreER activity after 4-OHT injection by crossing Nkx2-5(CreER/+) mice with a Rosa26(tdT/+) reporter strain...
August 2017: Genesis: the Journal of Genetics and Development
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