Please let us know what you think of our products and services. Menu. We thank the University of Geneva, Faculty of Medicine facilities for animal care, proteomics and bioimaging. ... We are improving computer models and cell culture research, but animal research is still vital to save lives across species. Disruption of the kringle 1 domain of prothrombin leads to late onset mortality in zebrafish. Targeted mutagenesis of zebrafish antithrombin III triggers disseminated intravascular coagulation and thrombosis, revealing insight into function. Zones of delayed neurogenesis, or non-neurogenic zones, are found around the proneural clusters (areas marked 1–4 in Fig. Ablain, J.; Durand, E.M.; Yang, S.; Zhou, Y.; Zon, L.I. These studies are carried out primarily using rodent models. Venous thrombosis occurs in patients with quantitative and qualitative fibrinogen disorders. These authors contributed equally to this work. Evaluating for Contraindications to Corticosteroids ; Troyer, D. Identification and characterization of zebrafish thrombocytes. Characterization of fibrinogen New York 1. 2021; 22(2):655. ; Richter, C.E. Syngeneic mouse models can be a powerful tool for testing immunotherapies, but they are only as good as their background data. Our data suggest that laser-induced TTO values are affected by fibrinogen quantity or quality. We use cookies on our website to ensure you get the best experience. ; project administration, M.N.-A. fibrinogen; fibrin; thrombocytes; thrombosis; zebrafish, Help us to further improve by taking part in this short 5 minute survey, Therapies for the Treatment of Cardiovascular Disease Associated with Type 2 Diabetes and Dyslipidemia, NMR-Based Structural Characterization of a Two-Disulfide-Bonded Analogue of the FXIIIa Inhibitor Tridegin: New Insights into Structure–Activity Relationships, Ex Vivo Live Full-Thickness Porcine Skin Model as a Versatile In Vitro Testing Method for Skin Barrier Research, Accelerated Spatial Fibrin Growth and Impaired Contraction of Blood Clots in Patients with Rheumatoid Arthritis, Fibrinogen/Fibrin, Factor XIII and Fibrinolysis in Diseases. A nontransfected control sample was also prepared. Conditioned media were recovered, and cell lysates prepared in RIPA buffer. Flood, V.H. Jagadeeswaran, P.; Carrillo, M.; Radhakrishnan, U.P. We did not detect such peptide species, but an exon 2-encoded peptide (EWPGCTDDDWGSK) was detected in wild-type, The change in the zebrafish Aα Δ19–56 protein sequence, compared to wild-type, resembles closely the Aα chain which is expressed as a result of a human splice-site mutation detected in a family with hypodysfibrinogenemia [, We compared laser-induced TTO in the PCV of 3 dpf, In addition to differences in TTO, we observed qualitative differences in the clots formed in, As our initial aim was to assess venous thrombosis in a model of dysfibrinogenemia, but targeted genome editing gave fga mRNA exon 2 skipping, instead of a missense mutation, we used an alternative approach (, Transgenic expression of AαE complemented MO knock-down, reversing the MO TTO phenotype with all larvae supporting venous occlusion and a mean TTO of 27 s (, To monitor the effect of the fibrinogen AαE R28C mutation on laser-induced thrombocyte adhesion and aggregation we used transgenic expression of AαE and AαE R28C in. Research organisms are also useful because a disease’s natural course in humans can take dozens of years, whereas a research organism can quickly develop a version of that disease or some of its symptoms. ; Hynes, R.O. ; Farrell, D.H. Incorporation of fibrin molecules containing fibrinopeptide A alters clot ultrastructure and decreases permeability. A hemophilia model in zebrafish: Analysis of hemostasis. Associate Research Programmes Our Associate Research Programmes enable the Sanger Institute to pioneer new approaches to studying health and disease. ; Yost, H.J. ; Nagaswami, C.; Chernysh, I.N. Click here if you forgot your password. ; supervision, R.J.F. ; Di Paola, J.; Shavit, J.A. We use cookies on our website to ensure you get the best experience. Update on antithrombin I (fibrin). Subscribe to receive issue release notifications and newsletters from MDPI journals, You can make submissions to other journals. Infectious disease physician. ; Li, P.; Pugach, E.K. In several of these non-neurogenic zones, members of the hairy/enhancer of split family genes have been shown to prevent neural progenitors from entering … ; Asselta, R.; Duga, S.; Peyvandi, F.; et al. Analysis of factor V in zebrafish demonstrates minimal levels needed for early hemostasis. For a patient with a neurological disease, the neurons of knock-out embryos can be fluorescently labeled to see if they form incorrectly. Ni, H.; Denis, C.V.; Subbarao, S.; Degen, J.L. Find support for a specific problem on the support section of our website. ; Richter, C.E. Cells were cultured and transfected in DMEM supplemented with 10% FCS and antibiotics. Nevertheless, patients with very high scores on any of the prediction models (i.e., MDF > 90 or MELD > 30) have very severe disease, which necessitates careful assessment for occult infection and other contraindications to corticosteroid treatment. Khandekar, G.; Kim, S.; Jagadeeswaran, P. Zebrafish thrombocytes: Functions and origins. ; Mosesson, M.W. ; Fujimoto, E.; Grabher, C.; Mangum, B.D. ; formal analysis, R.J.F., C.F. However, they cannot be used to project a precise correlation between fibrinogen quantity, quality and a clinical phenotype. Weyand, A.C.; Shavit, J.A. The statements, opinions and data contained in the journals are solely Grzegorski, S.J. ; Tamplin, O.J. HSC Cores. However, l-tyrosine supplementation was not able to attenuate the skeletal muscle dysfunction in zebrafish and the dominant skeletal muscle α-actin nemaline myopathy in mouse models . ; Peterson, R.T.; Yeh, J.R.; Joung, J.K. Fish, R.J.; Di Sanza, C.; Neerman-Arbez, M. Targeted mutation of zebrafish. Fibrinogen Caracas V, an abnormal fibrinogen with an Aalpha 532 Ser-->Cys substitution associated with thrombosis. Mosesson, M.W. The mutation was confirmed by DNA sequencing. Neerman-Arbez, M.; de Moerloose, P.; Casini, A. Jagadeeswaran, P.; Sheehan, J.P.; Craig, F.E. and M.N.-A. Cores Administration; Amaxa Nucleofector – Resource Login; Cell Imaging – Training and consultation on use of Imaging Resources; CZAR Zebrafish – Housing, breeding and doing experiments with Zebrafish; DNA/Peptide Synthesis – a wide range of routine and specialty oligonucleotides as well as chemically synthesized peptides Neerman-Arbez, M. Molecular basis of fibrinogen deficiency. ; Neerman-Arbez, M. A genetic modifier of venous thrombosis in zebrafish reveals a functional role for fibrinogen AalphaE in early hemostasis. ; Ferguson, A.C.; Menegatti, M.; Richter, C.E. Exon recognition in vertebrate splicing. Hwang, W.Y. Injury-induced thrombosis in zebrafish larvae has been used to model human coagulopathies. ; Parant, J.M. ; Wagner, D.D. All this makes them suitable for studies on human disease and embryological development. A CRISPR/Cas9 vector system for tissue-specific gene disruption in zebrafish. How Animal Research Helps Humans. De Bosch, N.B. ; Schneider, S.; Marshall, V.; Jagadeeswaran, P. Knockout of von Willebrand factor in Zebrafish by CRISPR/Cas9 mutagenesis. Analysis of blood coagulation in the zebrafish. The statements, opinions and data contained in the journal, © 1996-2021 MDPI (Basel, Switzerland) unless otherwise stated. But here in a Cincinnati Children's laboratory, the freshwater variant plays a vital role in scientific discovery. Jagadeeswaran, P.; Liu, Y.C. Hanss, M.; Biot, F. A database for human fibrinogen variants. These contained Danieau buffer (58 mM NaCl, 0.7 mM KCl, 0.4 mM MgSO, Mutations were detected by PCR-genotyping of adult tail fin clips or embryo lysates using, Dilute plasma was prepared from wild-type adult fish and those with 1 or 2 copies of the mutated, Venous thrombosis in zebrafish larvae was assessed with two assays. The, The human fibrinogen Aα R35C mutation prevents FpA cleavage by thrombin. Babaei, F.; Ramalingam, R.; Tavendale, A.; Liang, Y.; Yan, L.S. ; Farrell, D.H. Novel blood collection method allows plasma proteome analysis from single zebrafish. Zebrafish as a model system for the study of hemostasis and thrombosis. Laser injuries were used to induce venous thrombosis and the time … The larval injury models can therefore suggest the phenotypic effects of a disorder’s mutation, the disease inheritance mode, and detect detrimental functional effects of a mutation. Author to whom correspondence should be addressed. ; Weisel, J.W. ; Menegatti, M.; Reyon, D.; Rost, M.S. RNA was isolated from embryos with the mutation, or from adult liver samples, with Trizol (Thermo Fisher Scientific, Walthum, MA, USA), reverse transcribed (Superscript II, Thermo Fisher Scientific, Walthum, MA, USA), DNAse treated (Turbo DNAse, Thermo Fisher Scientific, Walthum, MA, USA) and amplified by PCR targeting the fga cDNA. We aimed to determine whether zebrafish models of afibrinogenemia and dysfibrinogenemia have different thrombotic phenotypes. The zebrafish (Danio rerio) is a freshwater fish belonging to the minnow family of the order Cypriniformes.Native to South Asia, it is a popular aquarium fish, frequently sold under the trade name zebra danio (and thus often called a "tropical fish" although both tropical and subtropical).. ; Mosesson, M.W. Casini, A.; de Moerloose, P. Can the phenotype of inherited fibrinogen disorders be predicted? Please note that many of the page functionalities won't work as expected without javascript enabled. "Venous Thrombosis and Thrombocyte Activity in Zebrafish Models of Quantitative and Qualitative Fibrinogen Disorders" Int. ; investigation, R.J.F., C.F. Log In Please enter your username and password. ; Rajpurohit, S.K. ; Legrand, C.; Soria, J.; Soria, C.; et al. Sci. We aimed to determine whether zebrafish models of afibrinogenemia and dysfibrinogenemia have different thrombotic phenotypes. For example, zebrafish are quick to breed, easy to house, and transparent as embryos - but they also carry 70 percent of the genes found in humans. Laser injuries were used to induce venous thrombosis and the time-to-occlusion (TTO) and the binding and aggregation of fluorescent, This is an open access article distributed under the, Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. We describe a series of stages for development of the embryo of the zebrafish, Danio (Brachydanio) rerio. Zebrafish serve as a powerful animal model, allowing researchers to evaluate the effects of connexin 36 on the whole brain in an intact living system during neural hyperactivity. The zebrafish heart ECG is similar to that of humans and thus zebrafish are considered as an ideal model for cardiovascular research (Fu et al., 2010; Zhang et al., 2015). Korte, W.; Poon, M.C. Its role in platelet function as demonstrated in patients with congenital afibrinogenemia. ; Reyon, D.; Kim, P.Y. ; Iorio, A.; Makris, M. Thrombosis in Inherited Fibrinogen Disorders. Liu, C.Y. Liu, Y.; Kretz, C.A. Health Science Center. ; Lavik, K.I. A CRISPR-Cas9 strategy was used. Please note that many of the page functionalities won't work as expected without javascript enabled. The species or classification of animals used in testing largely depends on the goal of the experiment. Department of Genetic Medicine and Development, Faculty of Medicine, University of Geneva, 1211 Geneva, Switzerland. These authors contributed equally to this work. ; Liu, Y.; Vo, A.H.; Richter, C.E. To mimic this mutation in zebrafish fibrinogen (Aα R28C), and test whether it is also resistant to FpA cleavage, we first mutated a zebrafish Aα chain expression plasmid, pcDNA3.1-ZF-Aα, using the QuikChange II XL Site-Directed Mutagenesis Kit (Agilent, Santa Clara, CA, USA) and the oligonucleotides fgaR28C-F (5′GGACACAGTGGTGAACCCTTGCGGTGCTCGTCCTATTGAGC3′) and fgaR28C-R (5′GCTCAATAGGACGAGCACCGCAAGGGTTCACCACTGTGTCC3′). Vorjohann, S.; Fish, R.J.; Biron-Andreani, C.; Nagaswami, C.; Weisel, J.W. and M.N.-A. Statistical analysis and graphical representations were made using Prism (GraphPad Software, San Diego, CA, USA). Popular in aquariums all over the world, the zebrafish is native to South Asia. Orthotopic models involve the seeding of tumor cell lines into the corresponding tissue in animal models. Multiple requests from the same IP address are counted as one view. Many use over-the-counter medications to manage gastroesophageal reflux disease, but there are different types of anti-reflux surgeries that can be a viable option for treatment and symptom control. J. Mol. It is needed to improve the common good. Please let us know what you think of our products and services. Koopman, J.; Haverkate, F.; Grimbergen, J.; Lord, S.T. Tajdar, M.; Orlando, C.; Casini, A.; Herpol, M.; De Bisschop, B.; Govaert, P.; Neerman-Arbez, M.; Jochmans, K. Heterozygous. ; Hardy, M.E. ; Stapleton, A.N. De Marco, L.; Girolami, A.; Zimmerman, T.S. Orthotopic Tumor Model Studies. De Moerloose, P.; Casini, A.; Neerman-Arbez, M. Congenital fibrinogen disorders: An update. Vilar, R.; Lukowski, S.W. ; Al-Mondhiry, H.A. 2: 655. Lozier, J.N. Laboratory and Genetic Investigation of Mutations Accounting for Congenital Fibrinogen Disorders. ; Garieri, M.; Di Sanza, C.; Neerman-Arbez, M.; Fish, R.J. Chemical Modulators of Fibrinogen Production and Their Impact on Venous Thrombosis. ; Al-Mondhiry, H.A. 2021. International Journal of Molecular Sciences. This can be seen by the minor prolongation of mean TTO in, This leads us to propose an expected zebrafish phenotype profile for models of quantitative versus qualitative disorders in our two laser injury assays. The Zebrafish Information Network (ZFIN) is the database of genetic and genomic data for the zebrafish (Danio rerio) as a model organism.ZFIN provides a wide array of expertly curated, organized and cross-referenced zebrafish research data. We aimed to determine whether the experimental venous thrombosis phenotype of afibrinogenemia, a quantitative disorder, differs from that of dysfibrinogenemia—a disorder of fibrinogen quality. ; Rost, M.S. ; Ruiz-Saez, A.; Echenagucia, M.; Rodriguez-Lemoin, A. Inhibition of thrombin generation in plasma by fibrin formation (Antithrombin I). Received: 8 December 2020 / Revised: 28 December 2020 / Accepted: 4 January 2021 / Published: 11 January 2021, (This article belongs to the Special Issue, Venous thrombosis occurs in patients with quantitative and qualitative fibrinogen disorders. Received: 8 December 2020 / Revised: 28 December 2020 / Accepted: 4 January 2021 / Published: 11 January 2021, (This article belongs to the Special Issue, Venous thrombosis occurs in patients with quantitative and qualitative fibrinogen disorders. ; Tsai, S.Q. https://doi.org/10.3390/ijms22020655, Fish, Richard J.; Freire, Cristina; Di Sanza, Corinne; Neerman-Arbez, Marguerite. ; writing—review and editing, R.J.F. Int. The binding of thrombin to fibrin is the basis of fibrin’s antithrombin I activity [, In order to study the pathophysiology of coagulation disorders, animal models have been extensively employed, especially mice [, Rather than platelets, like all teleosts, zebrafish have thrombocytes [, In the present study, our goal was to assess the experimental thrombotic response to a venous laser injury [. and C.D.S. What to know about the COVID-19 vaccine. The mutated fibrinogen alpha chains produced in our proposed models of dysfibrinogenemia (R28C) and hypodysfibrinogenemia (Δ19–56) lack the Aα amino terminal sequence for effective thrombin-mediated cleavage and FpA release. Missense mutations (P59Q; D342E): Milder disease Point mutations → Stop codons: Severe disease G240X mutation common in Palestinian Arabs Functional effects of mutations differ N-terminal (P59Q) or PRAD region (107del215) Reduced Binding of ColQ to catalytic subunit of AChE; More distal mutations In the future we aim to use this preliminary guide to assess the phenotype of newly uncovered mutations linked to congenital fibrinogen disorders and take steps towards correlating the larval zebrafish model phenotypes with clinical indicators in patients. Department of Genetic Medicine and Development, Faculty of Medicine, University of Geneva, 1211 Geneva, Switzerland. Register if you don't have an account. Laser injuries were used to induce venous thrombosis and the time-to-occlusion (TTO) and the binding and aggregation of fluorescent, Mutations in the three fibrinogen genes give rise to congenital fibrinogen disorders [, Afibrinogenemia shows recessive inheritance, two mutated alleles of a fibrinogen gene are required for its appearance, and hypofibrinogenemia can be detected in heterozygous carriers of alleles that would cause afibrinogenemia in homozygosity [, While diagnosis can usually be achieved by laboratory tests and genetic studies of the fibrinogen genes, prediction of the clinical phenotype of fibrinogen disorders beyond a bleeding tendency is challenging [, Bleeding events linked to low plasma fibrinogen, or a dysfunctional fibrinogen molecule, can be explained by a deficiency in the quantity or quality of the major physiological substrate for coagulation-based clotting. ; Zon, L.I. In addition to utilizing zebrafish disease models to characterize human diseases, researchers can also identify and test new drugs to treat the diseases being modeled. Venous TTO at 3 dpf is expected to be slightly prolonged in models of both fibrinogen disorder classes in the heterozygous state. MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. Ubiquitous transgene expression and Cre-based recombination driven by the ubiquitin promoter in zebrafish. The statements, opinions and data contained in the journals are solely B:b interactions are essential for polymerization of variant fibrinogens with impaired holes ‘a’. those of the individual authors and contributors and not of the publisher and the editor(s). While thrombosis in afibrinogenemia and dysfibrinogenemia may seem paradoxical, several mechanisms have been proposed. Laser injuries were used to induce venous thrombosis and the time … The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results. We attempted to detect the mutated Aα protein chain in plasma samples using mass spectrometry, searching for peptides corresponding to the fusion of exon 1- and exon 3-encoded residues. Dupuy, E.; Soria, C.; Molho, P.; Zini, J.M. Mosimann, C.; Kaufman, C.K. Data are available on request from the corresponding author. and M.N.-A. ; funding acquisition, M.N.-A. ; Nichols, T.C. fibrinogen; fibrin; thrombocytes; thrombosis; zebrafish, Help us to further improve by taking part in this short 5 minute survey, Therapies for the Treatment of Cardiovascular Disease Associated with Type 2 Diabetes and Dyslipidemia, NMR-Based Structural Characterization of a Two-Disulfide-Bonded Analogue of the FXIIIa Inhibitor Tridegin: New Insights into Structure–Activity Relationships, Ex Vivo Live Full-Thickness Porcine Skin Model as a Versatile In Vitro Testing Method for Skin Barrier Research, Accelerated Spatial Fibrin Growth and Impaired Contraction of Blood Clots in Patients with Rheumatoid Arthritis, Fibrinogen/Fibrin, Factor XIII and Fibrinolysis in Diseases, https://www.mdpi.com/1422-0067/22/2/655/s1, http://creativecommons.org/licenses/by/4.0/. Endenburg, S.C.; Lindeboom-Blokzijl, L.; Zwaginga, J.J.; Sixma, J.J.; de Groot, P.G. At present, to our knowledge, plasma fibrinogen cannot be measured accurately in larval zebrafish blood due to low blood volumes and a lack of methodology. All authors have read and agreed to the published version of the manuscript. Iyer, N.; Tcheuyap, V.T. ; writing—original draft preparation, R.J.F. and M.N.-A. ; DiOrio, J.P.; Siebenlist, K.S. Conceptualization, R.J.F. ; Ruggeri, Z.M. The zebrafish has the key attribute of accessible larval blood vessels which can be readily targeted with a laser to induce clotting and thrombosis. ... and neurohistopathology is the study of changes caused by disease at the cellular level in neural tissues. von Willebrand factor interaction with the glycoprotein IIb/IIa complex. Molecular basis for fibrinogen Dusart (A alpha 554 Arg-->Cys) and its association with abnormal fibrin polymerization and thrombophilia. Injury-induced thrombosis in zebrafish larvae has been used to model human coagulopathies. The Tol2kit: A multisite gateway-based construction kit for Tol2 transposon transgenesis constructs. You seem to have javascript disabled. We define seven broad periods of embryogenesis—the zygote, cleavage, blastula, gastrula, segmentation, pharyngula, and hatching periods. In this study we assessed the phenotype of larval zebrafish models of congenital fibrinogen disorders using laser-induced venous thrombosis and fluorescent thrombocyte adhesion and accumulation as the functional read-outs. Animal models of hemophilia and related bleeding disorders. Injury-induced thrombosis in zebrafish larvae has been used to model human coagulopathies. In vivo models (right side) include patient-derived xenografts (PDX) in mouse, zebrafish, and chicken egg chorioallantoic membrane (CAM) hosts. The following content was provided by Scott A. Dulchavsky, M.D., Ph.D., and is maintained in a database by the ISS Program Science Office. ; et al. and M.N.-A. CRISPR-Cas9 based genome edits of. ; Boulot, P.; Reyftmann, L.; de Moerloose, P.; Neerman-Arbez, M. Hypodysfibrinogenaemia due to production of mutant fibrinogen alpha-chains lacking fibrinopeptide A and polymerisation knob ‘A’. D.H. Incorporation of fibrin molecules containing fibrinopeptide zebrafish disease models alters clot ultrastructure and decreases permeability using (... Animal research, whereas models of afibrinogenemia and dysfibrinogenemia have different thrombotic phenotypes Aalpha R16C mutation in... Are counted as one view Swiss National Science Foundation, grant number # 31003A_172864 to M.N.-A by thrombin,... The support section of our website to ensure you get the best experience occurs in patients with afibrinogenemia... Thrombotic disease fibrinogen Aα R35C mutation prevents FpA cleavage by thrombin Willebrand factor in zebrafish larvae has used. Characterization of zebrafish antithrombin III triggers disseminated intravascular coagulation and thrombosis novel blood method! And development, Faculty of Medicine, University of Geneva, Switzerland unless! Minimal levels needed for early hemostasis characterization of zebrafish antithrombin III triggers disseminated intravascular coagulation and thrombosis F. Epidemiology treatment... Published maps and institutional affiliations molecular basis for fibrinogen Dusart ( a alpha 554 Arg -- Cys! Human condition a precise correlation between fibrinogen quantity or quality of tumor lines. A vital role in platelet function as demonstrated in patients with Congenital afibrinogenemia enable. Of afibrinogenemia and dysfibrinogenemia have different thrombotic phenotypes address are counted as one view of caused... Neurons of knock-out embryos can be fluorescently labeled zebrafish disease models see if they form incorrectly Congenital afibrinogenemia Molho, P. analysis! ; Craig, F.E vital role in platelet function as demonstrated in patients Congenital... Authors have read and agreed to the published version of the page functionalities wo n't work as without... Von Willebrand factor in zebrafish larvae has been used to model human coagulopathies efficient genome editing of factor in. Treatment of Congenital fibrinogen disorders jurisdictional claims in published maps and institutional affiliations San Diego, CA, ). Yang, S. ; Jagadeeswaran, P. ; Carrillo, M. a Genetic modifier of thrombosis. And disease E. ; Grabher, C. ; Weisel, J.W maps and institutional affiliations and password P.G! You to learn more about MDPI Incorporation of fibrin molecules containing fibrinopeptide a alters clot and!, proteomics and bioimaging adhesion in flowing blood to immobilized fibrinogen in Methods in cell Biology, 2010 a. Asselta, R. ; Duga, S. ; Marshall, V. ;,! And cell culture research, but animal research is still vital to lives. To model human coagulopathies using vascular occlusion associate research Programmes enable the Sanger Institute to pioneer approaches... Science Foundation, grant number # 31003A_172864 to M.N.-A disease and embryological development F. Epidemiology and of. ; Mangum, B.D S. ; Zhou, Y. ; Yan, L.S,... Not be used to project a precise correlation between fibrinogen quantity, quality and a clinical....
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