4B). In the study by Hong and colleagues, on-target and off-target effects could be … In light of recent results, they’re not so sure. The immunohistochemical results of the dystrophin expression showed that the chemical treatment did not restore dystrophin expression. However, PDE3 inhibitors, enoximone and milrinone (Fig. These fish are also small enough to be permeable to small molecules and can be assayed in large numbers. Treatment with 108 of the chemical pools was not lethal to the fish. While such screens show promise, using the correct model for screening is critically … Overall, this study highlights the predictive value of the zebrafish larva as a drug screening model for SCI. Red bars, chemically treated fish with 108 chemical pools (in duplicate). Other PDE5 and PDE4 inhibitors also had an effect in high concentrations (20 μg/mL). The treated dystrophin-null fish with aminophylline have normal structure in skeletal muscle. Three of seven candidate chemicals restored normal birefringence and increased survival of dystrophin-null fish. (C) Dystrophin-null fish. Immunohistochemical staining of whole-mutant and morphant fish bodies was done as described in ref. A survey of the existing literature from 2000 to 2017 identified 114 zebrafish chemical screens. They also have a primary organ system very similar to humans. 3), and indeed, some were toxic to all fish. Keywords: zebrafish, drug screening, compound screening, genetic kidney disease, high-throughput, high-content, automated microscopy INTRODUCTION Modern genetic diagnostics allow the rapid discovery of human disease-associated mutations. (B) Immunostaining of WT, nontreated, and chemical 4-treated fish with anti-myosin heavy chain and anti-laminin antibody. However, the homozygous dystrophin-null fish that showed no abnormality of birefringence showed no immunoreactivity with anti-dystrophin, and the same lack of staining was found in the nontreated dystrophin-null fish (Fig. The most common form of muscular dystrophy is Duchenne muscular dystrophy (DMD), representing more than 90% of the diagnosed cases. the zebrafish can serve as an early preclinical drug screening tool and can also help personalize cancer therapy by providing real-time data on the response of the human cells to treatment. All restored fish with chemicals 1–7 showed no expression of dystrophin, similar to those of untreated dystrophin-null fish (C). Those treated dystrophin-null fish with normal birefringence did not restore dystrophin expression (Fig. This library contains 1,120 small molecules composed of 90% marketed drugs and 10% bioactive alkaloids or related substances. analyzed data; and G.K. and L.M.K. All sequencing was accomplished in the Intellectual and Developmental Disabilities Research Center Molecular Core Laboratory supported by National Institute of Child Health and Human Development Grant 2P30HD018655-26. The small molecules used here were from the Prestwick library of bioreaticve human-approved use compounds. Light blue, nontreated WT fish. In the experimental groups tested with the chemical pools, the distribution of the affected fish ranged between 2.5% and 27.5%; 6 groups had less than 7.5% of mutant fish showing abnormal birefringence, and 16 groups had less than 10% (Fig. In the untreated dystrophin-null fish, the muscle fiber structure was disturbed, and some parts were abnormal (Fig. Twenty embryos from matings of heterozygous sapje fish are treated with a series of PDE inhibitors from 1 to 4 dpf (in triplicate at varying concentrations). However, treatment with each of the seven chemicals reduced the percentages of affected fish showing abnormal birefringence (Fig. Fish with nontreatment (A and B) and chemical treatment (C and D). Muscular dystrophy is a disease in which the muscle forms normally at first but then degenerates faster than it can be repaired. Some chemical pools decreased the percentage of affected fish showing abnormal birefringence compared with untreated fish and might contain a compound able to restore normal muscle structure. Lateral view of AP stained embryos at 72 h pf. Zebrafish platform Austria for preclinical drug screening (ZANDR) ZANDR was established at the Children´s Cancer Research Institute (CCRI) in 2019 and is a unique platform to phenotypically screen small compounds on zebrafish models of human diseases. Chemical screening in a zebrafish model of doxorubicin-induced cardiotoxicity identified the small molecules visnagin and diphenylurea, which rescued doxorubicin-induced cardiotoxicity in zebrafish and mice; visnagin bound to mitochondrial malate dehydrogenase (MDH2), and several MDH2 inhibitors exhibited similar cardioprotective effects, supporting visnagin and modulators of MDH2 activity as potential options for treating doxorubicin-induced … Long-term treatment of dystrophin-null fish selected to have abnormal birefringence at 4 dpf with chemicals 1–7 from 4 to 30 dpf (n = 3). Contributed by Louis M. Kunkel, February 10, 2011 (sent for review November 23, 2010). In 20 embryos, five apparently affected fish are caused by a homozygous mutation in the zebrafish dystrophin gene, which is inherited in a recessive manner (25%; arrows). To whom correspondence should be addressed at Drug Safety Evaluation, Research Laboratory for Development, Shionogi and Co., Ltd., 3-1-1 Futaba-cho, Toyonaka, Osaka 561-0825, Japan. Potential approaches to overcoming the hurdles to drug discovery in the face of complex biology are discussed, including in vivo screening of zebrafish … In fact, zebrafish … Isolation of candidate cDNAs for portions of the Duchenne muscular dystrophy gene, A cDNA clone from the Duchenne/Becker muscular dystrophy gene, PTC124 targets genetic disorders caused by nonsense mutations, Functional improvement of dystrophic muscle by myostatin blockade, Functional amounts of dystrophin produced by skipping the mutated exon in the mdx dystrophic mouse, Local dystrophin restoration with antisense oligonucleotide PRO051, Zebrafish: A model system for the study of human disease, Chemical discovery and global gene expression analysis in zebrafish, Small molecules that delay S phase suppress a zebrafish bmyb mutant, Small molecule developmental screens reveal the logic and timing of vertebrate development, The zebrafish as a model for muscular dystrophy and congenital myopathy, Delta-sarcoglycan is required for early zebrafish muscle organization, The dystrophin associated protein complex in zebrafish, Genetic isolation and characterization of a splicing mutant of zebrafish dystrophin, Genes required for functional glycosylation of dystroglycan are conserved in zebrafish, Developmental defects in a zebrafish model for muscular dystrophies associated with the loss of fukutin-related protein (FKRP), Zebrafish models for human FKRP muscular dystrophies, Zebrafish orthologs of human muscular dystrophy genes, Sildenafil reverses cardiac dysfunction in the mdx mouse model of Duchenne muscular dystrophy, Sarcolemma-localized nNOS is required to maintain activity after mild exercise, Theophylline: New perspectives for an old drug, Effects of aminophylline on cytokines and pulmonary function in patients undergoing valve replacement, Anti-inflammatory effects of low-dose oral theophylline in atopic asthma, Anti-oxidative effects of theophylline on human neutrophils involve cyclic nucleotides and protein kinase A, Primary role of functional ischemia, quantitative evidence for the two-hit mechanism, and phosphodiesterase-5 inhibitor therapy in mouse muscular dystrophy, An anchored PKA and PDE4 complex regulates subplasmalemmal cAMP dynamics, Protein kinase A signalling via CREB controls myogenesis induced by Wnt proteins, Convergent regulation of skeletal muscle Ca2+ channels by dystrophin, the actin cytoskeleton, and cAMP-dependent protein kinase, Stages of embryonic development of the zebrafish, Proceedings of the National Academy of Sciences, Earth, Atmospheric, and Planetary Sciences, www.pnas.org/lookup/suppl/doi:10.1073/pnas.1102116108/-/DCSupplemental, Science & Culture: At the nexus of music and medicine, some see disease treatments, News Feature: Tracing gold's cosmic origins, Journal Club: Friends appear to share patterns of brain activity, Transplantation of sperm-producing stem cells. (E) Distribution of the percentage of affected fish treated with chemical pools and untreated fish. The larval zebrafish xenograft assay has unique characteristics that make it unlike all other ex vivo assays and a top choice for in vivo drug screening. Zebrafish are increasingly swimming into the view of large-scale drug screening projects. Amanda Rodewald, Ivan Rudik, and Catherine Kling talk about the hazards of ozone pollution to birds. For drug screening using kcna1 morphants, 5 days post-fertilization (dpf) larval zebrafish maintained in 96-well plates were habituated for 20 min, under ambient light. These seven chemicals were divided into different groups: an antiinflammatory agent (chemical 1, epirizole), antiallergic agents (chemicals 2 and 3, homochlorcyclizine-dihydrochloride and conessine), a phosphodiesterase (PDE) inhibitor (chemical 4, aminophylline), an estradiol steroid (chemical 5, equilin), a chelating agent (chemical 6, pentetic acid), and a cardiotonic glycoside (chemical 7, proscillaridin A) as shown in Table 2. Zebrafish embryos were collected and raised at 28.5 °C according to standard procedures (30) and standard criteria (31) under the guidelines of our Institutional Animal Care and Use Committee. The authors declare no conflict of interest. In addition, we are grateful to Chris Lawrence and Jason Best, who managed our fish facility, and the Institute of Chemistry and Cell Biology at Harvard Medical School. 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