We investigated the mechanisms of intermediate cell mass (ICM) expansion in zebrafish chordin (Chd) morphant embryos and examined the role of BMPs in relation to this phenotype. At 24 h post-fertilization (hpf), the expanded ICM of embryos injected with chd morpholino (MO) (Chd MO embryos) contained a monotonous population of hematopoietic progenitors. In situ hybridization showed that hematopoietic transcription factors were ubiquitously expressed in the ICM whereas vascular gene expression was confined to the periphery. BMP4 (but not BMP2b or 7) and smad5 mRNA were ectopically expressed in the Chd MO ICM. At 48 hpf, monocytic cells were evident in both the ICM and circulation of Chd MO but not WT embryos. While injection of BMP4 MO had no effect on WT hematopoiesis, co-injecting BMP4 with chd MOs significantly reduced ICM expansion. Microarray studies revealed a number of genes that were differentially expressed in Chd MO and WT embryos and their roles in hematopoiesis has yet to be ...
The zebrafish genome contains ten genes that encode class II cytokine-like peptides, of which the two that are related most closely to mammalian interferon gamma (IFN-γ) were named IFN-γ1 and IFN-γ2. Although the zebrafish has become a popular model system to study immune mechanisms, and although interferons are central regulators of immunity, which zebrafish cytokines correspond functionally to mammalian IFN-γ has not been established. We used zebrafish embryos to assay the functions of IFN-γ1 and IFN-γ2, and have identified a subset of zebrafish homologs of the mammalian IFN-responsive genes as IFN-γ targets in the zebrafish embryo: these genes are upregulated in response to raised levels of either IFN-γ1 or IFN-γ2. Infection studies using two different pathogens show that IFN-γ signalling is required for resistance against bacterial infections in the young embryo and that the levels of IFN-γ need to be regulated tightly: raising IFN-γ levels sensitizes fish embryos against ...
J Vis Exp. 2012 Dec 16;(70). pii: 4243. doi: 10.3791/4243. Chang JT, Sive H. Source Department of Biology, Whitehead Institute of Biomedical Research, Massachusetts Institute of Technology. Abstract Cerebrospinal fluid (CSF) is a protein rich fluid contained within the brain ventricles. It is present during early vertebrate embryonic development and persists throughout life. Adult CSF is thought to cushion the brain, remove waste, and carry secreted molecules(1,2). In the adult and older embryo, the majority of CSF is made by the choroid plexus, a series of highly vascularized secretory regions located adjacent to the brain ventricles(3-5). In zebrafish, the choroid plexus is fully formed at 144 hours post fertilization (hpf)(6). Prior to this, in both zebrafish and other vertebrate embryos including mouse, a significant amount of embryonic CSF (eCSF) is present . These data and studies in chick suggest that the neuroepithelium is secretory early in development and may be the major source of ...
Danio rerio (zebrafish) have rapidly gained popularity in the study of vertebrate development and gene function. Zebrafish are an excellent model for performing large-scale genetic and drug screens to be performed coupled with the visualization of downstream perturbations on live, developing organisms. Due to the fact that large numbers of embryos are required to perform these experiments, we investigated ways to efficiently increase zebrafish embryo production while not being overly invasive or using chemicals that might affect the developmental processes. To perform these experiments, we mated wild type (wt) zebrafish while exposing them to the different wavelengths of light. Fish were exposed to 12 h of dark, followed by 4 h with the different colored light treatments. Once this light cycle was over, embryo production was enumerated. Results indicated that exposing zebrafish to the different wavelengths of fluorescent and LED light had little effect on embryo production. Further research may
Associate Professor, Biology Department. Syracuse University (Syracuse, New York). Service: I have been actively involved in the zebrafish community for my whole career. Ive attended all but one of the international zebrafish meetings since my first international zebrafish conference in 1996, all of the strategic conferences for zebrafish investigators held in the US and a couple of the European PI meetings. Recently I hosted and co-organized (with Jeff Amack and Jason Myers) the first ever Central New York zebrafish conference. I have been a PhD examiner for many zebrafish theses, an ad hoc reviewer for zebrafish grants for 12 different funding agencies in the US, Europe and Hong Kong and an NSF panel member for IOS Neural Organization. I was an elected committee member for the British Society for Developmental Biology for 5 years and was a scientific organizer for the annual BSDB conference in 2010 and a Frontiers of Science Symposium co-funded by the Royal Society (UK) and Japanese Society ...
Welcome to the Zebrafish Welfare Terms Wiki. The zebrafish welfare terms WIKI provides a standardised approach to the language used across facilities when describing zebrafish characteristics, which have the potential to impact welfare. We hope that by using these standardised terms we raise awareness of potential health concerns specific to genetic background and environmental impacts, which affect the well-being of zebrafish in our facilities and help bridge communication between different constituents involved in animal care. The terms have been created, reviewed, and added to by experienced animal caretakers and veterinarians.. If you would like to add to or edit this document please email one of these contacts. Further information regarding zebrafish welfare terms can be found in: Standardised Welfare Terms for the Zebrafish Community (http://zfin.org/ZDB-PUB-160421-6).. Goodwin et al_zeb_2016_1248.pdf. ...
The Chi-Bin Chien Award was established by the zebrafish research community and the International Zebrafish Society (IZFS) in memory of Dr. Chi-Bin Chien (1965-2011). Chi-Bin was Professor of Neurobiology and Anatomy at the University of Utah and served the international zebrafish community in numerous ways, including his service as Director of the Zebrafish Neural Development and Genetics Course at the Woods Hole Marine Biological Laboratory and as an organizer of the International Conference on Zebrafish Genetics and Development. The Award celebrates Chi-Bins enthusiasm and love for the discussion of scientific ideas and the good things that grow out of interactions and open discourse among scientists, his efforts to mentor and support the development of young scientists, and the collaborative and generous spirit with which he contributed to advance research with the zebrafish. The award will be given to an outstanding graduate student, postdoctoral trainee, or recently appointed faculty ...
2. Center for Research Informatics, The University of Chicago, 5751 S. Woodlawn Avenue, Chicago, IL 60637, USA. Zebrafish excel as a model organism for developmental biology and disease modeling. Traditionally, laboratory zebrafish have been maintained as outbred populations with high genetic variability. Our recent work has examined the core MHC locus in zebrafish, where we found alternative pathways of antigen processing and presentation genes that are separated by 500 million years of evolution. Here we performed highcoverage genomic sequencing for two clonal lines of zebrafish, and one partially inbred zebrafish, to uncover additional sources of immune gene variation throughout these genomes. Pathway analysis identified immune genes as highly enriched among genes under positive selection, or associated with structural variation. Overall, zebrafish genomes are enriched by approximately 5 fold higher levels of variation compared with humans, including SNVs, small indels, and structural ...
INTRODUCTION. Zebrafish maintains a remarkably higher ability than mammals to repair complex tissues after injury, including the heart and the central nervous system. For this reason, zebrafish embryos and their stem cells have been increasingly studied to unravel the molecular mechanisms underlying such regenerative potential, or to dissect evolutionary conserved pathways that may account for the regenerative action afforded by stem cells across different species.. Transplantation of human cord blood-derived CD34+ (hCD34+) cells into pregastrulation zebrafish embryos revealed that these human cells cosegregated with presumptive zebrafish hemangioblasts, being involved in early development of the embryonic vasculature of the recipient 1. Conversely, postgastrulation transplant resulted in the recruitment of hCD34+ cells into developing vessels, where their biology was mainly shifted to a paracrine action 2. These human cells were also found to accelerate vascular repair in adult zebrafish, after ...
MicroRNAs are a well-studied class of non-coding RNA and are known to regulate developmental processes in eukaryotes. Their role in key biological processes such as vasculature development has attracted interest. However, a comprehensive understanding of molecular regulation of angiogenesis and vascular integrity during development remains less explored. Here we identified miRNAs involved in the development and maintenance of vasculature in zebrafish embryos using a reverse genetics approach. Using a combination of bioinformatics predictions and literature based evidences we mined over 701 Human and 329 Zebrafish miRNAs to derive a list of 29 miRNAs targeting vascular specific genes in zebrafish. We shortlisted eight miRNAs and investigated their potential role in regulating vascular development in zebrafish transgenic model. In this screen we identified three miRNAs, namely miR-1, miR-144 and miR-142a-3p that have the potential to influence vascular development in zebrafish. We show that miR-142a-3p
The present study focuses on a cDNA fragment of 2.6 kb coding for glial fibrillary acidic protein (GFAP) related gene in zebrafish. Earlier work in the lab revealed that this gene showed 72% homology, at the amino acid level, with the mammalian gene coding for GFAP. Due to variations observed in sequence of head domain of GFAP and in expression pattern compared to that of rodent GFAP, this gene was named as zebrafish GFAP like gene (zfgfap-l)( Gene Bank Accession No: AY 397679).In this project, the expression pattern of zfgfap-l was studied by RT-PCR and in situ hybridization. RT-PCR results showed that the expression of zfgfap-l started as early as 1.5 hours post-fertilization (hpf) and increased steadily up to 10 hpf, then became a constant level of expression till 30 hpf. In embryos at sphere stage, the expression was detected in the cells of the superficial layer by in situ hybridization. After gastrulation, the expression became restricted to the neural tube, particularly in the presumptive ...
THE zebrafish (Danio rerio) is a powerful tool for understanding vertebrate biology. The usefulness of this model organism is bolstered by the availability of a "finished" sequenced and annotated genome (Howe et al. 2013; Flicek et al. 2014). As a natural extension of this resource, there are several high-throughput efforts to systematically mutagenize all zebrafish protein-coding genes (Moens et al. 2008; Kettleborough et al. 2013; Varshney et al. 2013a,b).. In addition to such projects, the combination of a sequenced genome and developments in targeted nuclease technology mean that the zebrafish community is now able to rapidly take advantage of custom genome-editing technologies (Doyon et al. 2008; Bedell et al. 2012; Hruscha et al. 2013; Hwang et al. 2013; Jao et al. 2013). CRISPRs in particular provide an efficient, easy, and inexpensive means of manipulating and interrogating the genome (Jinek et al. 2012; Cong et al. 2013; Mali et al. 2013). However, because there are very few hardy ...
RUNX1/AML1/CBFA2 is essential for definitive hematopoiesis, and chromosomal translocations affecting RUNX1 are frequently involved in human leukemias. Consequently, the normal function of RUNX1 and its involvement in leukemogenesis remain subject to intensive research. To further elucidate the role of RUNX1 in hematopoiesis, we cloned the zebrafish ortholog (runx1) and analyzed its function using this model system. Zebrafish runx1 is expressed in hematopoietic and neuronal cells during early embryogenesis. runx1 expression in the lateral plate mesoderm co-localizes with the hematopoietic transcription factor scl, and expression of runx1 is markedly reduced in the zebrafish mutants spadetail and cloche. Transient expression of runx1 in cloche embryos resulted in partial rescue of the hematopoietic defect. Depletion of Runx1 with antisense morpholino oligonucleotides abrogated the development of both blood and vessels, as demonstrated by loss of circulation, incomplete development of vasculature ...
Huang H., Lu F.I., Jia S., Meng S., Cao Y., Wang Y., Ma W., Yin K., Wen Z., Peng J., Thisse C., Thisse B., Meng A.. Angiomotin (Amot), the founding member of the Motin family, is involved in angiogenesis by regulating endothelial cell motility, and is required for visceral endoderm movement in mice. However, little is known about biological functions of the other two members of the Motin family, Angiomotin-like1 (Amotl1) and Angiomotin-like2 (Amotl2). Here, we have identified zebrafish amotl2 as an Fgf-responsive gene. Zebrafish amotl2 is expressed maternally and in restricted cell types zygotically. Knockdown of amotl2 expression delays epiboly and impairs convergence and extension movement, and amotl2-deficient cells in mosaic embryos fail to migrate properly. This coincides with loss of membrane protrusions and disorder of F-actin. Amotl2 partially co-localizes with RhoB-or EEA1-positive endosomes and the non-receptor tyrosine kinase c-Src. We further demonstrate that Amotl2 interacts ...
Purpose: Differentiation of diverse photoreceptor phenotypes in the vertebrate retina requires multiple signaling pathways that activate cascades of gene expression. The signaling molecule retinoic acid (RA) is known to regulate rod and cone cell fate, differentiation, and survival. The purpose of the current study is to identify photoreceptor genes controlled by RA signaling in the embryonic retina of the zebrafish.. Methods: We treated embryos with RA at 48 hours post-fertilization (hpf) and isolated total RNA from eyes for microarray analysis at 75 hpf in order to identify genes responding to RA over the period of photoreceptor differentiation. Differentially expressed genes were validated by quantitative RT-PCR (qRT-PCR) and in situ hybridization. Wild-type zebrafish and those carrying an RA signaling reporter transgene (RARE:YFP) were used.. Results: We identified 180 genes with significantly altered gene expression. Of interest was the long wavelength sensitive opsin 1 (LWS1) gene, which ...
The early axon scaffolding in the embryonic vertebrate brain consists of a series of ventrally projecting axon tracts that grow into a single major longitudinal pathway connected across the midline by commissures. We have investigated the role of Brother of CDO (BOC), an immunoglobulin (Ig) superfamily member distantly related to the Roundabout (Robo) family of axon-guidance receptors, in the development of this embryonic template of axon tracts in the zebrafish brain. A zebrafish homologue of BOC was isolated and shown to be expressed predominantly in the developing neural plate and later in the neural tube and developing brain. Zebrafish boc was initially highly localized to discrete bands in the mid- and hindbrain, but, as the major brain subdivisions emerged, it became more evenly expressed along the rostrocaudal axis, particularly in dorsal regions. The function of zebrafish boc was examined by a loss-of-function approach. Analysis of embryos injected with antisense morpholinos designed against boc
Midline convergence of organ primordia is an important mechanism for shaping the vertebrate body-plan at various stages of development, such as the morphogenesis of the heart and endoderm. Down regulation of wnt or noncanonical wnt signalling components, such as dishelleved (Dvl) or RhoA GTPase (RhoA), impairs midline convergence of the heart primordia and endoderm in zebrafish. This suggests that wnt signaling plays an important role in regulating midline convergence. At the early patterning stage of the zebrafish kidney, the two pronephric-glomerular primodia (PGP), which derive from intermediate mesoderm, converge towards the midline and fuse to form a functional pronephros. In contrast, during development of the mammalian kidney, the pronephros degenerates as the mesonephros develops without midline convergence. The hypothesis is thus that there is/are mechanisms underlying midline convergence of PGP in zebrafish, which is/are in addition to the control of the non-canonical wnt/Dvl/RhoA ...
The development of new sequencing technologies permits the study of mRNA expression to an unprecedented level. Recent RNA-seq experiments have analysed the changing transcriptome in the early zebrafish embryo (Aanes et al., 2011; Vesterlund et al., 2011; Pauli et al., 2012), but the work described here provides the first systematic identification of maternal and paternal mRNAs in this species. Previous microarray analyses had suggested that transcription occurs before the MZT in the zebrafish (Mathavan et al., 2005) and RNA-seq experiments have also described changes in mRNA levels prior to the MZT, suggesting that such changes were due to the post-transcriptional regulation of mRNAs (Aanes et al., 2011). By identifying zygotically expressed genes (Fig. 2) and then applying tight temporal resolution (Fig. 5B,C), our work confirms that, consistent with previous radioactivity incorporation experiments (Kane and Kimmel, 1993), zygotic transcription begins after ten cell cycles. Through the ...
Hundreds of loci have been robustly associated with circulating lipids, atherosclerosis and coronary artery disease; but for most loci the causal genes and mechanisms remain uncharacterized. The overall aim of my thesis is to develop and validate novel in vivo model systems that are suitable for high-throughput, image-based genetic screens in coronary artery disease and related traits, and use these model systems to systematically characterize positional candidate genes.. In Study I, I developed an experimental pipeline to validate the suitability of zebrafish larvae as a model system for systematic, large-scale characterization of drugs and genes associated with dyslipidemia and atherosclerosis. Using this pipeline, I showed that five days of overfeeding and cholesterol supplementation have independent pro-atherogenic effects in zebrafish larvae, which could be diminished by concomitant treatment with atorvastatin and ezetimibe. CRISPR-Cas9-induced mutations in orthologues of proof-of-concept ...
Hi Jorge, AnaSpec sells rabbit polyclonal anti-SOX9 antibody (cat# 28138) which will work for zebrafish SOX9a. Warm regards, Daniel [Zbrafish] Antibody for SOX9a Zebrafish Jorge Soares via zbrafish%40net.bio.net (by j.s.soares from gmail.com) Wed Aug 1 08:07:04 EST 2007 a.. Next message: [Zbrafish] PH meter b.. Messages sorted by: [ date ] [ thread ] [ subject ] [ author ] -------------------------------------------------------------------------------- Hi everyone, Just recently joined this mailing list, mainly because Im searching for an Antibody (mono or poli) that binds to SOX9a from Zebrafish, still leaving the TFBS (transcription Factor Bionding Site) open for it to be able to link to the DNA im studying. Does anyone know of a company that produces it? Cheers. Jorge -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20070801/9cfd411f/attachment.html -- I am using the free version of SPAMfighter. We are a ...
including: Anesthesia and Euthanasia in Zebrafish Guidance on the housing and care of zebrafish Danio Rerio 2011 Katso my s kaloista yleens : NC3Rs - Fish ...
We present a method to collect cerebrospinal fluid (CSF) and to create a system which lacks CSF within the embryonic zebrafish brain...
Proteoglycans (PGs) modulate numerous signaling pathways during development through binding of their glycosaminoglycan (GAG) side chains to various signaling molecules, including fibroblast growth factors (FGFs). A majority of PGs possess two or more GAG side chains, suggesting that GAG multivalency is imperative for biological functions in vivo. However, only a few studies have examined the biological significance of GAG multivalency. In this report, we utilized a library of bis- and tris-xylosides that produce two and three GAG chains on the same scaffold, respectively, thus mimicking PGs, to examine the importance of GAG valency and chain type in regulating FGF/FGFR interactions in vivo in zebrafish. A number of bis- and tris-xylosides, but not mono-xylosides, caused an elongation phenotype upon their injection into embryos. In situ hybridization showed that elongated embryos have elevated expression of the FGF target gene mkp3 but unchanged expression of reporters for other pathways, ...
Due to its transparency, virtually every brain structure of the larval zebrafish is accessible to light-based interrogation of circuit function. Advanced stimulation techniques allow the activation of optogenetic actuators at different resolution levels, and genetically encoded calcium indicators report the activity of a large proportion of neurons in the CNS. Large datasets result and need to be analyzed to identify cells that have specific properties - e.g. activity correlation to sensory stimulation or behavior. Advances in three-dimensional functional mapping in zebrafish are promising; however the mere coordinates of implicated neurons are not sufficient. To comprehensively understand circuit function, these functional maps need to be placed into the proper context of morphological features and projection patterns, neurotransmitter phenotypes, and key anatomical landmarks. We discuss the prospect of merging functional and anatomical data in an integrated atlas from the perspective of our work on
Recently, chemokine gradients have been directly visualized in vivo in the context of Ccl21‐mediated dendritic cell migration in mice (Weber et al, 2013) and in zebrafish, where Cxcl8 mediates neutrophil migration (Sarris et al, 2012) (Fig 2B). Cxcl8 forms an extracellular, matrix‐bound gradient that extends at least 100 μm around the cell that expresses the chemokine. Interestingly, Cxcl8 protein was detected beyond this local tissue gradient and was found to be enriched along the venous vasculature, which includes the CHT from which Cxcl8 meditates the mobilization of neutrophils into the vasculature (Sarris et al, 2012). Since Cxcl8 binding to the venous vasculature is also required for neutrophil arrest on the blood vessel wall and to facilitate the subsequent extravasation (Middleton et al, 1997), it appears that Cxcl8 acts at several stages of neutrophil recruitment to sites of infection.. Binding of Cxcl8 to the extracellular matrix, or more specifically to heparan sulfate ...
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A transcriptomics-based hepatotoxicity comparison between the zebrafish embryo and established human and rodent in vitro and in vivo models using cyclosporine A, amiodarone and ...
Health, ...Research has shown that mutations in the psen1 gene are common in the ...Professor Pertti Panulas research team at the University of Helsinki ...In the study the zebrafish was used as a model organism. The rapidly d...The lack of Presenilin-1 protein induced a change in the behavior of t...,The,zebrafish,revealed,a,central,regulator,for,the,development,of,the,brain,histamine,system,medicine,medical news today,latest medical news,medical newsletters,current medical news,latest medicine news
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Hi, I want to seperate leukocytes from whole blood and do cytospins. I am having problems with cells lysing. I was wondering if anybody knows the osmolarity of zebrafish blood and if there is a recommended PBS, HBSS... for blood cells of zebrafish. Any ideas? Thanks ...
The EZRC archives zebrafish lines and provides biomedical researchers in Europe with fish, plasmids, sequencing, genetic and chemical screening services.
Lens development involves interactions with other tissues. Several zebrafish eye mutants are characterized by an abnormally small lens ...
The zebrafish possesses all of the interleukin 2 receptor family except interleukin 2 receptor alpha and removal of the common signalling component interleukin 2 receptor gamma causes a human like severe combined immunodeficiency ...
(2011) Ding et al. PLoS ONE. Screening and evaluating anti- hepatitis C virus (HCV) drugs in vivo is difficult worldwide, mainly because of the lack of suitable small animal models. We investigate whether zebrafish could be a model organism for HCV replication. To achieve NS5B-dependent replicati...
Methods in Cell Biology, Volume 59 - The Zebrafish: Biology H. William Detrich III (Editor), Monte Westerfield (Editor), Leonard I. Zon (Editor), Leslie Wilson (Series Editor)
Harvesting lab-raised zebrafish based on their size led to differences in the activity of more than 4,000 genes, as well as changes in allele frequencies of those genes, in the fish that remained.. 0 Comments. ...
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J:138147 Gros-Louis F, Kriz J, Kabashi E, McDearmid J, Millecamps S, Urushitani M, Lin L, Dion P, Zhu Q, Drapeau P, Julien JP, Rouleau GA, Als2 mRNA splicing variants detected in KO mice rescue severe motor dysfunction phenotype in Als2 knock-down zebrafish. Hum Mol Genet. 2008 Sep 1;17(17):2691-702 ...
Zebrafish have been widely used as a model system for studying developmental processes, but in the last decade, they have also emerged as a valuable system for modeling human disease. The development and function of zebrafish organs are strikingly similar to those of humans, and the ease of creating mutant or transgenic fish has facilitated the generation of disease models. Here, we highlight the use of zebrafish for defining disease pathways and for discovering new therapies. ...
Burnett KG, Bain LJ, Baldwin WS, Callard GV, Cohen S, Di Giulio RT, Evans DH, Gomez-Chiarri M, Hahn ME, Hoover CA, Karchner SI, Katoh F, MacLatchy DL, Marshall WS, Meyer JN, Nacci DE, Oleksiak MF, Rees BB, Singer TD, Stegeman JJ, Towle DW, Van Veld PA, Vogelbein WK, Whitehead A, Winn RN, Crawford DL. Fundulus as the premier teleost model in environmental biology: opportunities for new insights using genomics. Comparative Biochemistry and Physiology Part D: Genomics and Proteomics 2007;2(4):257-286 ...
CALM modulates theformation of tau-positivetangles in zebrafish.(a) Histological section to demonstrate the individual cell layers ofthe zebrafish retina. The p
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Nothing that sparkly should be food safe, it just isnt natural. Granted, it does make the fish shimmer in a semi-realistic--and somewhat menacing--way. Like one of those frogs or insects that use brightly colored markings to indicate (or mimic) toxicity and deter predators from eating them. I found myself reluctant to take a bite ...
The goal of this research is to apply an innovative technology for gene knockout to identify and understand developmental effects of drugs and other chemicals i...
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Weissman-Unni, along with Ian Lake B.A. 13, Leah Weston 14, and Kyla Hamling 14 (seated), work in the microinjection area within her research lab. Hamling uses a special dissection microscope to view zebrafish embryos ...
The advent of high-throughput sequencing has greatly accelerated the identification of inherited and de novo disease causing mutations. Following discovery the analysis of the developmental and cellular pathways of the affected genes is a crucial step on the path towards therapy with model organisms as the central tool. Traditionally, vertebrate model organisms such as mouse and zebrafish have been used on a gene-by-gene basis, however, in order to keep pace with the increasing speed of discovery, new approaches are needed. Owing to a high quality genome reference sequence and its genetic and embryological tractability the zebrafish is a vertebrate model especially suited for large scale studies. Previously having established methods to generate and identify disruptive zebrafish point mutations on a genome wide scale the Zebrafish Mutation Project is now assigning biological function to every protein-coding gene in the zebrafish genome. We submit alleles to a high-throughput assessment of ...