Herkenhoff, marcos e oliveira, arthur c nachtigall, pedro g costa, juliana m campos, vinicius f. Mar 29, 20 mir145 and mir143 were downregulated during odontoblast differentiation. Development of zebrafish epidermis weijen chang and pungpung hwang zebrafish epidermal ionocytes are analogous to mammalian kidney cells in terms of expression and function of ion transporters. Dissection of organs from the adult zebrafish protocol.
Micrornas function to regulate the expression levels of other genes by several mechanisms. Ascomycota, the dominant phylum in the intestine of zebrafish. Unfortunately, traditional murine models are not efficient for the further study of ibd. Biological functions of mir590 and its role in carcinogenesis. Host gut motility promotes competitive exclusion within a.
Childs1 biochemistry and molecular biology, and smooth muscle research group, university of calgary, calgary, ab, canada t2n 4n1. Remarkable epigenetic modifications and reprogramming of gene expression occur to activate the embryonic genome. Pdf mir145 directs intestinal maturation in zebrafish. Organisms can be protected from bleach exposure when sequestered within. We discuss the potential role of mir 143 145 as valuable biomarkers for cardiovascular diseases and explore the potential strategy of targeting mir 143 and mir 145. Frontiers the intestinal mycobiota in wild zebrafish. Zebrafish are an excellent model for the study of intestinal immunity. S1 a with increasingly specific expression in the heart, ear, and pharyngeal. Reverse genetics screen in zebrafish identifies a role of mir. Jun 9, 2010 fatima macho sanchezsimon, xiao xiao zhang, horace h. Biology, husbandry, diseases, and research applications is a comprehensive work that fulfills a critical need for a thorough compilation of information on this species. A guide for the laboratory use of zebrafish danio rerio, 3rd ed university of oregon, eugene, or. Repression of the mir143145 cluster by oncogenic ras.
Development and growth of organs in living whole embryo. The cardiovascular system is critical for the survival of vertebrates, and is one of the earliest organ systems to develop in an embryo. Currently there is no treatment for microsporidiosis in zebrafish. Although experimental analysis of the embryo and larva is extensive and the morphology has been well documented, descriptions of adult zebrafish anatomy and studies of development of the adult structures and organs. We are beginning to understand the functions of mirna145 played during essential biological processes. Establishment of intestinal homeostasis during the. The expression pattern of grna is similar to that previously reported 35. In zebrafish embryos, intestinal tract compartmentalizes into three segments. Loss of mir145 leads to defects in heart and gut development.
We visualize, in zebrafish, the effects on epithelial barrier function and intestinal autophagy of onecourse and repetitive injury. Proceedings of the national academy of sciences of the united states. To determine the role of fgf signaling in the differentiation of zebrafish intestinal cells, we treated wild type embryos with su5402 and used the tghsp70l. It is known that cellular differentiation is governed by mirnas, for example it was recently shown that mir145 directs the intestinal development in danio rerio. Zebrafish danio rerio is a fresh water fish that inhabits rivers in india, pakistan and other places in asia. Repetitive injury induces increased mortality, impaired recovery of intestinal barrier function, failure to contain bacteria within the intestine. As a clinically frequent disease, it can affect individuals throughout their lives, with multiple complications. A procedure for bleaching embryos can be found in the zebrafish book westerfield, 2000, which is available online through zfin, and is distributed in print by zirc. Thyroid hormone th signaling promotes tissue maturation and adult organ formation. An exclusive cellular and molecular network governs intestinal smooth muscle cell differentiation in vertebrates.
The rapid specification and differentiation of the embryonic zebrafish gut is essential to provide contractility for the digestion of food. Proceedings of the national academy of sciences of the united states of america. Throughout early embryonic development, a differential methylation pattern is maintained although. Peristaltic motility and mass transport are, of course, key attributes of animal intestinal tracts. Patterning mechanisms of the sub intestinal venous plexus in zebrafish. Regulation of microrna during cardiomyocyte maturation in sheep. Recent studies have identified engrailed2 en2, a homeoboxcontaining transcription factor, as a candidate oncogene in prostate cancer pc. This regulatory task is of critical importance to prevent inappropriate immune activation that may otherwise lead to. Periodic health reports zirc public wiki zebrafish. To ascertain whether mir145 is expressed in the mouse placenta and fetus to determine if mir145 is associated with placental growth in mice by examining both placenta growth and mir145 expression across gestation.
Cytogenetics, fluorescent in situ hybridization fish, and microarray techniques are discussed in chapter 2. Therapeutic targeting on en2, however, is limited because the mechanism underlying en2 overexpression in prostatic cancer cells is unknown. Microrna145 mir145 is considered to play key role in many cellular processes, such as proliferation, differentiation and apoptosis by inhibiting target gene expression. Product description type unit quantity academic price book. The intestinal mucosa faces the challenge of regulating the balance between immune tolerance towards commensal bacteria, environmental stimuli and food antigens on the one hand, and induction of efficient immune responses against invading pathogens on the other hand. Genetic analyses and gene targeting experiments in zebrafish offer a novel approach to dissect such interactions at a molecular level. Microrna1423p mir1423p is considered to play key role in many cellular processes, such as proliferation, differentiation and apoptosis by inhibiting target gene expression. They are encoded within the genes that code for the smspecific contractile proteins, are under the transcriptional control of myocardin, and are therefore highly enriched in smcs, with negligible. Recently, zebrafish has sparked interest in other branches of the biomedical research.
Ascomycota, the dominant fungal phylum in the intestine of zebrafish, is the largest and wide spread phylum of the kingdom fungi. Zebra fish mirnas, dremir15a2 and dremir16c, orthologous to the q14. Thus, effective and convenient animal models are needed. At a higher taxonomic level phylum, we did not observe any differences between the samples from wildcaught and laboratoryreared fish. Mir145 1,747 words exact match in snippet view article find links to article carter ad, childs sj oct 2009. Fluorescent in situ hybridization results of mir145 a and mir143 b during the differentiation of primarily cultured odontoblasts at day 0 d0, day 5 d5, and day 9 d9.
We are taking a genetic approach to identify new genes involved in angiogenesis, the process by which new blood vessels develop. Here we show that the microrna mir145 is strongly expressed in zebrafish gut smooth muscle and regulates its development. Because of the robust activation of mir145 by n1icd, we utilized this method of notch activation. They are encoded within the genes that code for the smspecific contractile proteins, are under the transcriptional control of myocardin, and are therefore highly enriched in smcs, with negligible expression in other cell types. Reverse genetics screen in zebrafish identifies a role of mir 142a3p in vascular development and integrity. The mir143145 cluster is a novel transcriptional target of. We used the zebrafish as a model to test whether and how th signaling affects pancreatic islet maturation, and consequently glucose homeostasis. Microrna143 and 145 in colon cancer dna and cell biology.
Their cotranscription means they are frequently studied together in the same cellular pathways and diseases. The role of micrornas in modulating gut epithelial or smooth muscle differentiation is currently not known. The zebrafish atlas of macroscopic and microscopic anatomy the zebrafish danio rerio is avaluable and common model for researchers working in the fields of genetics, oncology, and developmental sciences. Publications the childs laboratory university of calgary. The smooth muscle microrna mir145 regulates gut epithelial development via a paracrine mechanism. Thyroid hormone coordinates pancreatic islet maturation. Regulation of microrna during cardiomyocyte maturation in sheep article pdf available in bmc genomics 161. The transcriptional regulation of cardiovascular development requires precise spatiotemporal control of gene expression. Deciphering the porcine intestinal microrna transcriptome.
Hepatotoxicity of palmitic acid in zebrafish involves the. Klf5 is a documented target of mir145 via sequences in the 3. Two of the zebra fish orthologues fk54b05, fw91a08 for the human genes arghap20 and fdx from the 11q2223 deleted region also lie on the same region of zebra fish chromosome 9. S1a with increasingly specific expression in the heart, ear, and pharyngeal arches between 24 and 72 hpf. In early zebrafish development between 16 and 24 hpf, mir145 is expressed ubiquitously at very low levels fig. We are beginning to understand the functions of mirna1423p played during essential biological. Oxazoloneinduced intestinal inflammation in adult zebrafish. Expression of activated kras leads to downregulation of the mir 143 145 cluster in diverse model systems. This study was to investigate the potential regulatory role of mir 33a on en2 expression and explore this.
Moreover, monzo and colleagues found a correlation between the mirna expression profiles in human embryonic colonic mucosa and colorectal carcinoma. In the early stages of preimplantation embryo development, maternal mrnas direct embryonic development. The specific zirc bleaching procedure is also outlined in the protocols link from the zirc homepage. In order to identify mirnas regulated by kras signaling, we used a custom microarray to profile mirna expression in a nontransformed pancreatic ductal epithelial cell line hpne km lee et al. Intestinal development in amniotes is driven by interactions between progenitor cells derived from the three primary germ layers. In the past two decades, zebrafish has become one of the preferred in vivo model organisms for studying diverse processes like embryogenesis 8, 9, autophagy, behaviour 11, 12 and sleep. In mutant zebrafish that have reduced intestinal motility due to mutation of the gene ret, which impairs ens development and function, competition between these bacterial species can be neutralized. However, distinguishing and following cells over extended time periods remains difficult. Mir 590 is a member of microrna family, and its gene is located at 7q11.
Inflammatory bowel disease ibd is a chronic, recurrent, and remitting inflammatory disease with unclear etiology. Modulation of mir145 levels results in gut smooth muscle and epithelium maturation defects. Intestinal growth and differentiation in zebrafish. Where do i start zfin is the zebrafish model organism database and as such we work towards integrating zebrafish biology with its genome.
This fullcolor atlas will aid experimental design and interpretation in these areas by providing a fundamental understanding of zebrafish. Loss or gain of mir145 function phenocopies defects observed with altered gata6 expression and accordingly, we show that mir145 directly represses gata6, and that gata6 is a major mir145 target in vitro and in vivo. Over the last 20 years, the zebrafish has become a powerful model organism for understanding vertebrate development and disease. To uncover regulators of intestinal barrier function and inflammation in zebrafish, we performed a forward genetic screen to identify mutants with defects in gut epithelium integrity.
Mir 145 inhibit stem cell characteristics univerthabitat. Glial cells are important constituents of the nervous system and a. At 120 h postfertilization hpf, the gut epithelium in control wildtype wt siblings has a. Utr reporter to analyze mir145 functional activity. George sanders presentation occupational health and safety veterinary care. Fibroblast growth factor receptor 2c signaling is required. A read is counted each time someone views a publication summary such as the title, abstract, and list of authors, clicks on a figure, or views or downloads the fulltext. Among vertebrates, zebrafish is uniquely suited for in vivo imaging owing to its small size and optical translucency. For example, zebrafish oral intubation has been used in the study of mycobacterium marinum and mycobacterium peregrinum infection 1. Zebrafish modeling of intestinal injury, bacterial.
Mature mir 5903p and mir 5905p exist in the body and perform different functions by combining different target genes, and sequences of mir 590 are showed in fig. Results loss of mir145 leads to defects in heart and gut development. Using genetic lineage tracing, we demonstrate that ismcs arise from the lpm in a. The availability of several transgenic reporter fish for different innate and adaptive immune cells and the high homology in terms of gut function and morphology enables in depth analysis of the process of intestinal inflammation.
Developmental transitions alter an organisms metabolic requirements, and it remains unclear how development and metabolic demands are coordinated. Control should focus on optimizing husbandry conditions and removal of all emaciated and moribund fish to prevent cannibalism. Development and growth of organs in living whole embryo and larval grafts in zebrafish skip to main content thank you for visiting. Morphine regulates dopaminergic neuron differentiation via. Epigenetic control of intestinal barrier function and. Two micrornas, mir 145 and mir 143, have been shown to be present in multipotent cardiac progenitor cells in the mouse embryo. Advances in imaging and celllabeling techniques have greatly enhanced our understanding of developmental and neurobiological processes. Zebrafish gut as a house for human intestinal bacteria. In molecular biology, mir145 microrna is a short rna molecule that in humans is encoded by the mir145 gene. Therefore, a zebrafish vascular model was selected to evaluate the influence of hif2. A guide for the laboratory use of zebrafish danio rerio, eugene, university of. Zebrafish 101 for iacucs focus of monte matthews presentation institutional oversight training and education for both iacucs and zebrafish researchers when to have a protocol counting zebrafish focus of dr.
Application of zebrafish models in inflammatory bowel disease. After feeding for 5 d, there were significant differences in the relative mrna expression of trx and saa2 between gf zebrafish fed an hfd and a 12pa diet p development follows a series of critical events. Carter, evvilynn rollins, sonja georgijevic, massimo m. The text provides significant updates for working vivarium professionals maintaining zebrafish colonies, veterinarians responsible for their care and wellbeing, zoologists and.