e DMP1-Cre-ERT2 mice expression has been shown to be specific to osteocytes and a few mature osteoblasts. (36) Before beginning of tamoxifen treatment, body weight (Supporting Fig. 3A), BMD (Supporting Fig. 3B), bone mineral content (Supporting Fig. 3C), body lean mass (Supporting Fig. 3D), and fat mass (Supporting Fig. 3E) were Cited by: 35. We welcome your input and comments. Please use is form to recommend updates to e information in ZFIN. We appreciate as much detail as possible and references as appropriate. ZEBRAFISH meeting attendees will have e opportunity to present eir research rough oral and poster presentations and to hear e latest updates. ZEBRAFISH organizers are confident at we will offer a strong scientific programme, including many opportunities to learn, share, network and interact wi our sponsors at e virtual exhibition. 01, · Generation of Cre/loxP transgenic zebrafish for tracing tumor cell lineage To trace e fate of tumor hepatocytes, a Cre/loxP-based fate-tracing approach was adopted.As shown in Fig. 1A, a el transgenic line, Tg(fabp: loxP-EGFP-stop-loxP-DsRed. TRE: CreER T2) (abbreviated to CreER), was established and crossed wi e existing xmrk transgenic zebrafish to obtain e CreER/xmrk Cited by: 2. 30, · e meeting will begin wi dinner and an evening session on Wednesday ember 20, and conclude wi lunch on Saturday, ember 23, . is meeting will provide a forum for presentation and discussion of e most in ative and exciting research currently ongoing in neuroscience using e zebrafish model. e zebrafish genetics toolkit has been missing a particularly handy piece of kit: a promoter to drive ubiquitous transgene expression roughout development, equivalent to e Rosa26 locus used in mouse genetics. But no longer, for in one of Development's inural Technical papers (p. 169), Leonard Zon and co-workers report at e zebrafish ubiquitin (ubi) promoter can drive constitutive. Protein Leng Homeobox-like domain superfamily Paired-box protein 2 C-terminal Paired DNA-binding domain Paired domain PAX family Winged helix-like DNA-binding domain superfamily. 24, 2009 · Apoptosis plays important roles in embryogenesis, tissue homeostasis, and immune system regulation. e zebrafish (Danio rerio) is a powerful vertebrate model organism at has been extensively used to study apoptotic cell dea during normal development and under conditions of cellular stress. In e past 5 years, a detailed picture has begun to emerge of e molecular underpinnings of . Please acknowledge e Zebrafish International Resource Center in all publications resulting from e materials and/or services we are providing to you. ank you. Zebrafish International Resource Center (ZIRC) 5274 University of Oregon Eugene, OR 97403-5274, USA Phone: 541-346-6028. INTRODUCTION. A ubiquitous promoter/enhancer to drive transgenes is a key component of a complete transgenesis toolkit in any model organism. Enhancer trap screening in e mouse previously identified e Rosa26 locus, which became e current gold standard for ubiquitous transgene expression and for Cre/loxP-based lineage tracing at all stages of development and in all adult tissues. e DMP1‐Cre‐ERT2 mice expression has been shown to be specific to osteocytes and a few mature osteoblasts. 36 Before beginning of tamoxifen treatment, body weight (Supporting Fig. 3A), BMD (Supporting Fig. 3B), bone mineral content (Supporting Fig. 3C), body lean mass (Supporting Fig. 3D), and fat mass (Supporting Fig. 3E) were similar. To fur er confirm e specificity of is antibody, we analyzed a mouse spermatogonial stem cell line harboring floxed alleles for e Foxos and a tamoxifen-inducible Cre ERT2 transgene. e principal band disappeared following tamoxifen administration, confirming at is band corresponds to Foxo1 . col1a2 Cre-ERT2. ubi:Switch embryos were pulsed wi 4-OHT for 2 h at 3 dpf to mosaicly label col1a2 + MPCs, and e mosaicism allows for reliable cell tracing over multiple time points. At 24 h after e 4-OHT pulse, a small fraction of col1a2 + MPCs on e surface of e somite was labeled by e mCherry expression, but no myofibers were. Zebrafish can regenerate after brain injury, and e regenerative process is driven by resident stem cells. Stem cells are heterogeneous in e vertebrate brain, but e significance of having heterogeneous stem cells in regeneration is not understood. Limited availability of specific stem cells might impair e regeneration of particular cell lineages. We studied regeneration of e adult. To identify a cre-effector TAU line wi high efficiency of recombination, all F1 fish for TAU were outcrossed to Tg(her4.1:mCherry-t2a-Cre ERT2) or Tg(nbt:mCherry-t2a-Cre ERT2) cre-driver lines, and e F2 progeny were treated wi freshly prepared 5 µM of Tamoxifen (TAM) in E3 medium between 16–32 hpf and 32–48 hpf. At 48hpf, e F2. 08, · lymphatic and cardiac defects 1 mutants fail to form a lymphatic vasculature. In a screen for zebrafish lymphatic vascular mutants, we identified a mutant dubbed lymphatic and cardiac defects 1 (lyc1).lyc1 mutants exhibited a reduction or loss of e main axial lymphatic vessel, e oracic duct (TD) at 4 days post fertilization (dpf), as well as mild cardiac oedema, but retained blood. FIG URE 1 Generation of mouse line carrying e Reck CreERT2 allele. (a) Knock-in strategy. DTA, diph eria toxin fragment. FRT, FLP recombinase target. yellow box, poly-A signa. Fgf pa way expression in e adult neural retina. e role of Fgf signaling in e adult zebrafish retina is little studied, hence we initially investigated e expression profile of several Fgf receptors (Fgfr), ligands and target genes by in situ hybridization (Fig. 1 A–M).Reverse transcriptase (RT PCR analysis on cDNA prepared from adult zebrafish eyes revealed e presence of. 29, · Italian Zebrafish 2021-2022 (Feb. 4 and 11, 2021 - Webinars) SETAC Europe Session Fish model species in human and environmental toxicology (virtual. 3-6 2021) 16 International Zebrafish Conference (IZFC), hosted by e International Zebrafish Society (e 16 – 20, 2021 - Montréal, Québec, Canada). 20, · For UAS:Cre-ERT2 (Tg(14xUAS-E1b:BGi-Cre-ERT2-2a-Cer)y563) we first inserted Cer.zf1 and e rabbit β-globin intron into pT1UMP. We en amplified and inserted Cre-ERT2 from pCAG-Cre-ERT2 (gift from Connie Cepko, Addgene plasmid 14797), adding an in frame 2a sequence before Cerulean. 08, · Aldh1b1 Is a Specific Molecular ker of Centroacinar Cells. First, we analyzed in e pancreas of adult mice (8 to mo old unless o erwise stated) e coexpression of Aldh1b1 wi pancreas lineage and progenitor kers using a specific Aldh1b1 antibody (SI Appendix, Fig. S1 A and B).All ese Aldh1b1 + cells are epi elial (Fig. 1A and SI Appendix, Fig. S1C) wi invariably . Procedures for doing is are in e Zebrafish Book, A guide for e laboratory use of zebrafish (Danio rerio), Monte Westerfield, ed. available on-line or for a nominal fee you can order hard copies from e Resource Center. If you have not grown zebrafish before, you might want to practice before we ship you fish. An internal ribosome entry site (IRES) was inserted between GFP and Cre-ERT2 to create LV-Id1pr/p-GFP-IRES-Cre ERT2 (Id1pr/p-Cre ERT2). A constitutive Cre-ERT2 expressing control construct was created by replacing e Id1pr/p element wi an 800 bp region of e elongation factor (EF) 1α1 promoter to generate LV-pWPT-EF long -GFP- IRES -Cre. Studies of prostate cancer pa ogenesis and development of new erapies have been hampered by a lack of appropriate mouse models. We have generated PSA-Cre-ERT2 mice at express e tamoxifen-dependent Cre-ERT2 recombinase selectively in prostatic epi elium, us allowing us to target floxed genes selectively in epi elial cells of fully differentiated prostate of adult mice and to modulate. When ese mice are used in con ction wi a Cre recombinase-expressing strain, successful Cre-mediated excision results in e constitutive expression of e EGFP::L A fusion gene in Cre-expressing tissues. is strain allows identification of specific changes in mRNA populations by translating ribosomal affinity purification where recovery of intact cells for analysis is not possible. Hereditary hemorrhagic telangiectasia (HHT) is an autosomal dominant vascular disorder at leads to abnormal connections between arteries and veins termed arteriovenous malformations (AVM). Mutations in TGFβ pa way members ALK1, ENG and SMAD4 lead to HHT. However, a Smad4 mouse model of HHT does not currently exist. We aimed to create and characterize a Smad4 endo elial cell (EC) . Genetic labeling techniques allow for noninvasive lineage tracing of cells in vivo. Two-photon inducible activators provide spatial resolution for superficial cells, but labeling cells located. e PSA-Cre-ERT2 transgenic mouse strain expressing Cre-ERT2 under control of e promoter of e human PSA gene is described e consensus report from e Bar Harbor meeting of e Mouse Models of Human Cancer Consortium Prostate Pa ology Committee. Cancer Res., 64, Zebrafish miR-462-731 is required for digestive organ development. 08, · PCR assays for additional zebrafish pa ogens, such as Pseudocapillaria tomentosa, will be available soon. Samples are received by ZIRC and transferred to OVDL for appropriate testing. Mycobacteriosis. Diagnosis of mycobacteriosis and species identification of Mycobacterium spp. require different approaches compared to Gram negative bacteria. One vector uses e EF1A promoter to drive expression of Cre‐ERT2, followed by a P2A self‐cleaving peptide and en Turbo‐GFP. A second vector uses e EF1A promoter to drive expression of human ceramide syn ase 2 (CERS2) fused to a flexible linker peptide (3xGGGS) followed by mRFP1. von Hippel-Lindau (VHL) disease is an autosomal-dominant tumor predisposition syndrome characterized by e development of highly vascularized tumors and cysts. LOH of e VHL gene results in aberrant upregulation of hypoxia-inducible factors (HIF) and has been associated wi tumor formation. Hemangioblastomas of e central nervous system and retina represent e most prevalent VHL. 01, · Resulting offspring carrying e transgene (Pappa2 wt/fl. Cre-ERT2) were mated to mice heterozygous for e conditional allele (Pappa2 wt/fl) and e body weight and tail leng of offspring were measured at 3, 6, 14 and 18 weeks of age (N = –22 per genotype, wi 6 genotypes: ree Pappa2 genotypes, each wi or wi out e Cre-ERT2. Hans et al. show at, even at temperature ranges of 37–42°C, recombination events can be blocked completely in Tg(hsp70:Cre-ERT2) animals if tamoxifen is not applied following heat shock. O er intriguing developments include e generation of zebrafish wi mosaic expression of oncogenic transgenes [ 66, 67 ] allowing more detailed. 20, · Lineage tracing is e identification of all progeny of a single cell. Al ough its origins date back to developmental biology of invertebrates in e 19 century, lineage tracing is now an essential tool for studying stem cell properties in adult mammalian tissues. Lineage tracing provides a powerful means of understanding tissue development, homeostasis, and disease, especially when it. Apr 01, · An Emerging Role for e EphrinB2-EphB4-RASA1 Signaling Axis in Human Congenital Cerebrovascular Disorders. Normal vascular development includes de o blood vessel formation from endo elial cells (ECs) (vasculogenesis) 1, 2, expansion of e network rough vessel sprouting, splitting, and remodeling (angiogenesis), and differentiation of ese vessels into arteries, veins, and . Apr 06, · Adult zebrafish are often difficult to section wi out first softening eir skin and scales. e day before processing, transfer e fixed fish to 5 trichloroacetic acid (TCA) in Dietrich’s fixative, and place on rotor overnight. TCA will soften e scales . 17, · Bars represent means ± s.e.m. from n = 5 mT/mG Vav‐Cre ERT2 Chk1 fl/fl mice. E, F. (E) Percentage of living LK or LSK cells and (F) percentage of GFP + LK or LSK cells found in e bone row of mT/mG Vav‐Cre ERT2 (WT) and mT/mG Vav‐Cre ERT2 Chk1 fl/fl (Chk1 fl/fl) mice on day 8 or 15 after first TAM. Select a stage name below to get a detailed description and images. You can also browse e zebrafish Anatomical Ontology (AO) to show anatomical terms at are present at at stage. (h = hours of development at 28.5°C). Since e 1930s, zebra fish have been a model organism for studying human diseases. e fertilized eggs, embryos, and fry are transparent, allowing scientists to easily observe and study topics such as tumor grow, brain tissue development, and blood vessel grow. However, after a few weeks, transparency lines as eir bodies become opaque. Cre recombinase-controlled lox site recombination is a cornerstone of contemporary mouse genetics, and Cre/lox techniques erefore attract increasing interest in e zebrafish field. Feb 07, · e UBC-cre/ERT2.FLExDUX4 mice ubiquitously express a TMX-inducible cre from e human ubiquitin C promoter, and, in e absence of TMX administration, should not express levels of DUX4-fl mRNA above ose caused by antisense transcription of e inverted FLExDUX4 transgene. erefore, double transgenic animals were not anticipated to have any. RESULTS. e Noto-cre allele was created by knocking e site-specific recombinase gene cre into exon 2 of e Noto locus .Using is strategy, cre is expressed in all Noto-expressing cells, specifically in e node and posterior notochord between E7.5 and E12.5 (Yamanaka et al., 2007), and in all of eir descendents.To ensure at e temporal regulation of Noto expression was not altered. e salamander is e only tetrapod at regenerates complex body structures roughout life. iphering e underlying molecular processes of regeneration is fundamental for regenerative medicine and developmental biology, but e model organism had limited tools for molecular analysis. We describe a comprehensive set of germline transgenic strains in e laboratory-bred salamander Ambystoma. Genetic labelling of zebrafish CMs using a UV-inducible Cre/ lox recombination system. Double transgenic embryos (Tg(myl7:Cre-Ert2) Double transgenic embryos (Tg(myl7:Cre-Ert2) þ / 2. To be able to analyze e role of UTX in adult mice, we crossed e conditional UTX FD mice to e Rosa-Cre-ERT2 line. 29 e Cre-ERT2 fusion protein is expressed from e Rosa26 locus, but Cre-ERT2 associates wi e hsp90 in e cytoplasm and remains inactive. To induce Cre expression, mice were treated at 11 to 14 weeks of age wi. A subset of patients wi gastric cancer have mutations in genes at participate in or regulate Wnt signaling at e level of ligand (Wnt) receptor (Fzd) binding. Moreover, increased Fzd expression is associated wi poor clinical outcome. Despite ese findings, ere are no in vivo studies investigating e potential of targeting Wnt receptors for treating gastric cancer, and e specific. e Zebrafish International Resource Center is committed to working wi you to ensure successful transition of your valuable fish strain to ZIRC. We will work closely wi you roughout is process. If you have comments or suggestions, please use our online form or call (541) 346-6028. e small G-protein NRAS is mutated in 22 of human melanomas, whereas e related proteins KRAS and HRAS are mutated in only 2 and 1 of melanomas, respectively. We have developed a mouse model of melanoma in which Cre recombinase/ LoxP technology is used to drive inducible expression of G12VKRAS in e melanocytic lineage. e mice develop skin hyperpigmentation, nevi, and tumors .