Congress Abstracts - Society for Developmental Biology

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Program/Abstract # 590 In vivo imaging of Xenopus laevis development using an Ultra-Compact MRI Huebner, Kelli R. (Knox College, USA); McDowell, Andrew (ABQMR, Inc., USA); Thorn, Judith (Knox College, USA) Magnetic resonance imaging (MRI) is a noninvasive technique used to reconstruct a three dimensional image of an intact subject. MRI is necessary for in vivo imaging of internal development of Xenopus laevis embryos because they are opaque. An Ultra-Compact MRI (UC-MRI) was created specifically for imaging of X. laevis development in place of a superconducting MRI system which is both large and expensive for a research setting. The UC-MRI has successfully imaged various events in X. laevis development without affecting embryo viability. Germinal vesicle breakdown in a progesterone treated oocyte has been imaged over 13 hours. Early cell cleavage and gastrulation have been imaged in a time series both with and without a T1-weighted contrast agent— gadolinium chelated with diethylenetriaminepentaacetic acid (Gd-DTPA; MAGNEVIST). This identifies morphological changes to the blastocoel and epibolic movement of the ectodermal sheet. Furthermore, a chart of internal images throughout various stages of development which correlates to the external pictures of the Nieuwkoop and Faber series (1994) has been assembled. Current and future experiments involve manipulating the dorsal∕ventral axis patterning to see if failure to gastrulate can be detected internally before phenotypic changes occur and also in vivo fate mapping of tissues using magnetic nanoparticles. Program/Abstract # 591 Highly effective ex vivo gene manipulation to study kidney development using self-complementary adeno-associated viruses (scAAV) Zhou, Qin; Chen, Tielin; Wang, Honglian (West China Hospital, China); Gao, Guangping (U Mass Med Sch, USA) Ex vivo cultures of intact embryonic kidneys have become a powerful system for studying renal development. However, few methods are available for gene manipulation in those ex vivo intact tissue cultures and have impeded identification and studies of genes in this developmental process. Here we demonstrate that self-complementary adeno-associated virus(scAAV)is highly effective in delivering and expressing genes into the deep tissues of cultured kidney and that the effectiveness varies with different scAAV serotypes. These findings are expected to expedite and expand use of the ex vivo embryonic kidney cultures for kidney development research and our understanding of Kidney development. Program/Abstract # 592 Withdrawn Program/Abstract # 593 Development and evolution of vertebrate external genitalia Martin Cohn (U Florida, USA) Few morphological characters in the animal kingdom rival the diversity, complexity and evolvability of the external genitalia. Among animals with internal fertilization, genital morphology evolves rapidly, and the size, shape and anatomical details of the external genitalia can show dramatic variation, even between closely related species. External genital development begins with the initiation of paired genital swellings that merge to form the genital tubercle, the precursor of the penis and clitoris. While some aspects of an ancient appendage development network, including Shh, Fgf10-FgfR2, Wnt5a, Hoxa/d13 and Bmps, are involved in outgrowth and patterning of the genital tubercle, other genes known to play central roles in limb development, such as Fgf8, play no role in genital development. In addition to proximodistal outgrowth and dorsoventral patterning, the genital tubercle of mammals undergoes tubular morphogenesis of the endodermal epithelium to form the urethral tube. Formation of a closed urethral tube goes awry at a high frequency in humans, with incomplete closure (hypospadias) affecting ~1:250 live births. Urethral tube closure is a relatively recent evolutionary innovation that occurs only in mammals; in non-mammalian amniotes, the phallus forms an open groove (sulcus) rather than a tube. Turtles form such a phallus with a sulcus whereas squamate reptiles (snakes and lizards) have paired phalluses, known as hemipenes, situated on either side of the cloaca. In birds, the phallus was reduced or lost in ~97% of species. The developmental mechanisms responsible for the evolution of external genitalia are unknown. By integrating mouse developmental genetics with comparative studies of external genital development across a range of taxa, we are gaining new insights into the molecular mechanisms of external genital defects in mammals and the diversification of genital form during vertebrate evolution. Program/Abstract # 594 Cardiac gene regulatory networks in health and disease Romaric Bouveret, Mirana Ramialison, Reena Singh, Nicole Schonrock (Victor Chang Cardiac Res. Inst., Australia); Antoine Bondu (U Libre de Bruxelles, Belgium); Li Xin, Gavin Chapman, Sally L. Dunwoodie (Victor Chang Cardiac Res. Inst., Australia); Cedric Blanpain (U Libre de Bruxelles, Belgium); Richard P. Harvey (Victor Chang Cardiac Res. Inst., Australia) Our previous studies have identified a number of highly conserved transcriptional factors (TFs), signalling molecules and non-coding RNAs that are the controlling elements of the gene regulatory networks essential for heart specification, morphogenesis and function. However, the architecture of the cardiac gene regulatory networks as they evolve in time and space remains ill-defined. Progress in this area is essential for understanding metazoan biology and for realising the promise of new therapies for a host of diseases. The cisregulatory interactions between TFs and their target gene promoters/enhancers is a major determinant of network architecture. We 170
have begun to document the targets of a large number of developmentally-active and tissue-restricted cardiac TFs, and mutant versions, using the DamID technique and the HL-1 cardiomyocyte cell line, which beats homogeneously in culture. Our findings suggest that ubiquitous TFs are embedded within the cardiomyocyte network at an executive level, and that mutant TFs bind a surprising number of targets through co-factor interaction, including to hundreds of “off-targets” never seen by the wildtype factor and which may play a role in congenital heart disease pathology. Program/Abstract # 595 Defining mechanisms of imprinted expression at Igf2r/Airn during mouse gastrulation Marcho, Chelsea; Bevilacqua, Ariana; Veeramani, Swarna; Mager, Jesse (Univ of Massachusetts-Amherst, USA) Genomic imprinting is an epigenetic mechanism resulting in differential gene expression in a parent-of-origin manner. In the mouse genome, there are approximately 100 imprinted genes, many of which occur in imprinted gene clusters. DNA methylation and histone modifications have been shown to be allele specific at imprinted loci, corresponding with imprinted (allele specific) expression. At the Igf2r/Airn cluster (which contains 3 maternally expressed genes Igf2r, Slc22a2, and Slc22a3), the paternally expressed non-coding RNA Airn is thought to be responsible for silencing the paternal alleles of Igf2r, Slc22a2, and Slc22a3, resulting in reciprocal imprinting at the locus. Here we examined the changes in imprinted gene expression in a tissue-specific and time-dependent manner during gastrulation. We found that in embryonic tissue prior to gastrulation, Igf2r is biallelic and Airn is not expressed. Once gastrulation commences, Igf2r and Airn become reciprocally expressed and imprinted. To examine the epigenetic mechanisms involved in the establishment of Igf2r/Airn imprinting, we characterized changes in DNA methylation by bisulfite sequencing of two differentially methylated regions (DMR) at the locus. Consistent with ES cell models, we found spreading of DNA methylation at DMR2 during the start of gastrulation, corresponding to the changes in expression. Additionally, expression of CTCF, a factor involved in imprinting regulation at the H19/Igf2 locus, is only present once gastrulation begins, suggesting CTCF may also play a role in imprinting at Igf2r/Airn. Our data suggest a model similar to H19/Igf2 in which DNA methylation changes at onset of gastrulation, blocking the binding of the methylation sensitive factor CTCF, which allows paternal Airn expression and silencing of paternal Igf2r. Program/Abstract # 596 Vascular plastic response in the primary somatosensory cortex of birth-enucleated rats Silvia Zenteno De León, Raquel Martínez Méndez, Gabriel Gutiérrez Ospina (UNAM, México) Blind individuals display an expansion of the primary somatosensory cortex (S1) and increments of its vascular density. Even though both S1 expansion and increased vascular density in birth enucleated rats are thought to result from increased sensory experience, we have recently published evidence that sensory experience is not increased in birth-enucleated rodents (Fetter-Pruneda et al., 2013). The mechanisms leading to S1 vascular plasticity must then be carefully revised. We then evaluated vascular density in 7 days old (PD7) and adult rats enucleated at birth and in those having intact sight. Although we confirmed previous results showing increments in vascular density in the adult S1 of birth-enucelated rats, we were able to see such increments only in 20% of the sampled birth enucleated rats at PD7. Interestingly, this change occurs when all birth enucleated rats had already displayed an expanded S1. Our results thus support that increased vascular density is a consequence of S1 expansion in birth enucleated rats and that is not necessarily related with increments in the inflow of sensory experience. Financial aid came from CONACYT 82879. Program/Abstract # 597 Morphogenesis in the sea urchin: linking dynamically remodeling network states to protease function in development of skeletogenic and non-skeletogenic mesoderm Lyons, Deirdre C.; Dougherty, Mark; Saunders, Lindsay; McClay, David (Duke University, USA) In the sea urchin embryo 64 Primary Mesenchyme Cells (PMCs) synthesize a calcium carbonate skeleton that defines the shape of the larva. The PMC lineage comprises 4 cells at the vegetal pole in the early embryo. These cells then divide, invade the blastocoel via an epithelial-to-mesenchymal transition (EMT), and undergo directed migration in response to ectodermal cues. Next, PMCs fuse forming a syncytium within which the skeletal rudiment is secreted and subsequently elaborated by branching and elongation. Previous research established a gene regulatory network (GRN) for PMC specification. We focus on uncovering subroutines controlling morphogenetic processes. We and others identified subsets of transcription factors in the PMC GRN that are necessary for behaviors such as cell invasion, migration, cell fusion and skeletal patterning. Using GRN analysis and time-lapse imaging we connect these subnetworks to cell biological machinery proteins that carry out specific cellular processes. For example, metalloproteases are necessary for invasion, migration and cell fusion in many cell types in other systems. We find that several families of metalloproteases are dynamically expressed in the PMCs during EMT, migration and fusion, and are even expressed by certain PMC nuclei at the growing rod tips. These patterns suggest a highly regulated spatiotemporal program of extra cellular matrix (ECM) modification by the PMCs during successive step of differentiation. This work establishes the PMCs as a model for studying how a dynamically remodeling GRN launches successive cellular behaviors within a single cell type. Furthermore, protease expression is also observed in non-skeletogenic mesodermal cell types that are known to undergo later EMTs and remodel the ECM presumably to enable cell migration and tissue fusion. Treating embryos with MMP inhibitors causes the PMCs to fail to make skeleton, and prevents proper mouth formation and pigment cell differentiation, which involve non-skeletogenic mesoderm. These data suggest that proteases play a 171
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International Society of Developmen
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neural crest cells (hNCC)) human em
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activity may determine the orientat
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Developmental cell signaling pathwa
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opposed roles during early gastrula
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functions by the recruitment of add
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function validated with explant stu
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Program/Abstract # 48 Modeling regu
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surface of the embryo. In zebrafish
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morphologies. Our analyses not only
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junctional proteins (RPE patterning
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Program/Abstract # 78 In vivo recep
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Program/Abstract # 85 Guts and gast
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Program/Abstract # 92 The C. elegan
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Program/Abstract # 98 A gradient of
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Program/Abstract # 106 Developing a
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NSCs, but not in neurons, bind not
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abnormalities in the development of
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Tbx4 and contributes to Tbx4 repres
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downregulated by polycomb-group pro
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Zac1 expression in the developing c
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understanding the mechanisms that u
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Program/Abstract # 156 Systematic I
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Program/Abstract # 163 Size regulat
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TACC3 was localized around the DNA
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Penaeid shrimp represent an importa
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tubule. Using live imaging of cultu
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show that cell polarity is disrupte
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process. However, a clear mechanist
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Zhang, Haoran; Wong, Elaine Yee-man
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condition. Kanyon embryos have a mi
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Program/Abstract # 218 Lfng regulat
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works in concert with basal cues fr
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medaka genome. These miRs are encod
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facilitan cambio en patrón morfoge
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coordinately regulate the expressio
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downstream asymmetric signals. The
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viability and morphology of over 20
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owser. The genomic differences obse
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evolutionary analysis to study the
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teleostei. Our data evidenced that
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ontogeny. The typical condition for
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examples whose Shh expression requi
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insights into the ancestral role of
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Program/Abstract # 308 Mechanistic
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Patients with Joubert Syndrome and
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Program/Abstract # 323 Drosophila g
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signalling during gut and enteric n
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normally form, hence producing prox
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survival in Shh mutant embryos foll
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conserved in amniotes other than ch
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maturation and function. We strive
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Urness, Lisa D; Bleyl, Steven B (Un
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development is already unknown. Emb
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in A-P and P-D patterning by establ
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perturbed in arco piccolo mutants w
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Hoxb7-Cre line, leads to multiple u
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differentiation from common progeni
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investigate this issue, we used tar
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Page 133 and 134: Program/Abstract # 462 Retinaldehyd
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Congress Abstracts - Society for Developmental Biology

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