Congress Abstracts - Society for Developmental Biology

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was improved, and post-prandial glucose was lower. We observed that in biotin supplemented animals, the islet was closer to achieve the adult typical morphology due to a faster cell remodeling as well a major expansion rate of β cell mass. The augmented proportion of β cell mass is caused by the increase of proliferation and neogenesis. This remarks finding in our data suggest that biotin accelerates postnatal maturation of islet. Program/Abstract # 583 Drosophila survival can be altered by protein diet Pena Rangel, Maria Teresa; Riesgo, Juan (UNAM-Querétaro, Mexico) Regulation of metabolism is a central feature of life. This encompasses growth, development, sexual maturation, fertility, health, survival, and overall lifespan. Dietary composition is critical in the conformation of the ultimate organismal energy balance. In many organisms like C. elegans, D. melanogaster, mice, and primates feeding under caloric restriction that do not lead to overt malnutrition has been found to extend lifespan. Reduced insulin signaling can also increase the longevity in flies and rodents. Despite this, little has been done to test the effect on survival of diets with different amounts of proteins. To test the effects of dietary protein restriction on survival in wild type flies, we evaluated isocaloric normal and low protein diets with or without the addition of methionine using both molasses-yeast based standard food recipe, and a chemically defined fly diet formulation. Our results show increased survival with the addition of methionine in the low protein molasses-yeast diet. We are currently extending the effects of the same dietary conditions on mutant genotypes of the insulin-signaling pathway. Program/Abstract # 584 Influence of Dietary Minerals on Sex Determination of Mice Embryos Faqih, Reham; Alhimaidi, Ahmad (King Saud Univ, Saudi Arabia) The embryo sex preselection before pregnancy received considerable attention at the level of human and animal with a controversy over the years. The application of the embryo sex preselection production systems have become essential at the level of human and animal and increasingly for medical and economical interest. There are many ways to preselect the sex of the embryos, including specific meals and minerals which it is perfect because of its simplicity, low cost and ethical popular acceptance. The aim of this study is investigate of the influence of dietary minerals on sex determination of mice embryos. Mice food formula was prepared by ARASC animal food company, for male preselection (Na and K) was increased about 20% more than the regular or control mice diet, while for female preselection (Ca, Mg and P) were increased about 20%. The three experimental animal SWR/J mice (Male, female preselection and the control group) were contain 11 females, each 2or 3 females were housed with one male. The offspring data were collected, liver samples and blood samples were collected from each group. Data were satirically analyzed by GraphPad InStat Program soft wear. The result show a significant differences (P < 0.05) in the male preselection diet group (Na, K) they produce (53 males 62.6% ) compared to the female offspring (30 female 37.2%). While the data show no significant differences in the female or control groups. In the blood parameters the results also showed the existence of significant differences at (P
cycle, no changes were observed in expression of p21 and proliferating cell nuclear antigen (PCNA) after irradiation. Since that DNA lesion may be caused by different damaging agents, we investigate if lesions caused by UVB radiation activate the p53 pathway. UVB irradiated embryos showed an overexpression of p53. Our data indicates that after irradiation, embryos at E7 were not able to repair DNA damage, but may activate a possible apoptosis pathway. Program/Abstract # 587 Spectral confocal imagining with aberration correction as a tool for 3D rendering from whole mouse embryos Moody, Sally A.; Popratiloff, Anastas; Oakley, Beverly; Maynard, Thomas; Meechan, Daniel; Wang, Xin; Mendelowitz, David; LaMantia, Anthony (George Washington U, USA) Although confocal microscopy has been broadly used for acquiring 3D data sets from whole embryos, a number of barriers limit the utility of this approach. Tissue transparency, auto-fluorescence and spherical aberration are among the leading restrictive factors. In an effort to refine existing approaches, we combined: use of clearing agents; proper aberration correction; and use of spectral confocal imaging with linear unmixing. Using this combination of refined approaches we investigated the utility of 3D registration of confocal data sets from whole mouse embryos. E10-11 day mouse embryos double immunolabeled (ICC) for neurofilament and PCAM or embryos expressing endogenous GFP were used. ICC embryos were cleared with BABB (benzyl alcohol, benzyl benzoate, 1:2) and imaged from a glass depression chamber in BABB with coverslip. Fixed embryos expressing GFP were cleared with Scaleview A2 solution. GFP fluorescence was imaged from whole embryos with or without a coverslip. Imaging was performed with multiimmersion 25x/0.8 objective lens using Zeiss 710. Using spectral linear unmixing the auto-fluorescence was readily separated from Alexa Fluor 488 and 647, thus allowing for unobstructed visualization of the blood vessels with PCAM and cranial nerves with neurofilament antibodies. Using a matching refractive index between the point source and the objective lens minimized the spherical aberration and provided reliable 3D registration. A similar approach was taken to produce 3D sets of scale-cleared embryos. A combination of tile- and z-scanning allowed for generation of 3D data sets from large areas of the embryo, while also preserving the resolution at a fine scale including individual axons and growth cones. Program/Abstract # 588 Doxycycline-controlled and Recombinase-Activated Gene OverexpressioN (DRAGON): an intersectional strategy for targeting precise and reversible gene expression in mice Zeni, Eliane; Silva, Heloisa; Maia, Guilherme; Müller, Yara; Ammar, Dib; Nazari, Evelise (Univ Fedl de Santa Catarina, Brazil) The combination of tissue-specific Cre drivers and Cre-responsive mouse lines has allowed the development of sophisticated genetic approaches for studies of gene function in the mouse. However, it is very common that the cell subpopulation of interest is not defined by the expression of a single marker, but by a combination of at least two. Here we present the initial characterization of a strategy that utilizes a Rosa26 knock-in allele to express a gene only in cells defined by the combination of tissue-specific Cre activity and regulated (r)tTA activity. We name this system Doxycycline-controlled and Recombinase-Activated Gene OverexpressioN (DRAGON). This intersectional strategy achieves two important goals: 1) expression of a gene is restricted to the subpopulation of cells that expresses Cre and (r)tTA; and 2) temporal control of gene expression is provided via doxycycline administration, allowing a gene to be turned on or off at will. The DRAGON allele is also designed with fluorescent markers to identify cells that have undergone Cre recombination and in which (r)tTA is active, while a different fluorescent protein is expressed in the remaining (Cre–) cells in which (r)tTA is active, enabling the different cell populations to be characterized or isolated for further molecular analysis. With great utility, the DRAGON system takes advantage of the growing number of Cre(ER) and (r)tTA alleles available, with the potential for almost unlimited cell-type specific targeting. In conclusion, this approach should provide the opportunity to address biological questions in vivo that were previously hampered by technical limitations. Program/Abstract # 589 Chemical control of Wnt/b catenin signalling during development Gonzalez Malagon, Sandra Guadalupe; Liu, Karen (King’s College London, UK) The ability to manipulate protein function is very important when studying biological events. A classic approach is to genetically eliminate the protein of interest. Although “knockout” models have been most successful in biological research, there are drawbacks. For example, if the protein has a significant role at different stages of development, the knockout model may not survive to the stage of interest. To overcome this problem, alternative conditional strategies have been developed; but again, they are limited. Small molecules would be ideal, as these have the advantage of being reversible and can be administered at specific time points. Nevertheless, the huge challenge is to find small molecules that are specific, with minimal secondary effects. We are testing a novel approach that makes use of engineered chemically regulated destabilizing domains. These domains confer drug-dependence to a protein of interest. In the absence of drug, the protein is rapidly degraded. We are assessing the advantages of this technology by targeting the key players of the Wnt signalling pathway, including b-catenin and GSK3b, which are both essential during development. In this way we aim to develop a highly useful tool for biologists in diverse research fields that will provide the advantages of specific and reversible control of protein function. 169
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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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Page 133 and 134: Program/Abstract # 462 Retinaldehyd
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Congress Abstracts - Society for Developmental Biology

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