Driever, W. et al. A genetic screen for mutations affecting embryogenesis in zebrafish. Development 123, 37-46

Cardiovascular Research Center, Massachusetts General Hospital, Charlestown 02129, USA.
Development (Impact Factor: 6.46). 01/1997; 123:37-46. DOI: 10.5167/uzh-215
Source: PubMed


Systematic genome-wide mutagenesis screens for embryonic phenotypes have been instrumental in the understanding of invertebrate and plant development. Here, we report the results from the first application of such a large-scale genetic screening to vertebrate development. Male zebrafish were mutagenized with N-ethyl N-nitrosourea to induce mutations in spermatogonial cells at an average specific locus rate of one in 651 mutagenized genomes. Mutations were transmitted to the F1 generation, and 2205 F2 families were raised. F3 embryos from sibling crosses within the F2 families were screened for developmental abnormalities. A total of 2337 mutagenized genomes were analyzed, and 2383 mutations resulting in abnormal embryonic and early larval phenotypes were identified. The phenotypes of 695 mutants indicated involvement of the identified loci in specific aspects of embryogenesis. These mutations were maintained for further characterization and were classified into categories according to their phenotypes. The analyses and genetic complementation of mutations from several categories are reported in separate manuscripts. Mutations affecting pigmentation, motility, muscle and body shape have not been extensively analyzed and are listed here. A total of 331 mutations were tested for allelism within their respective categories. This defined 220 genetic loci with on average 1.5 alleles per locus. For about two-thirds of all loci only one allele was isolated. Therefore it is not possible to give a reliable estimate on the degree of saturation reached in our screen; however, the number of genes that can mutate to visible embryonic and early larval phenotypes in zebrafish is expected to be several-fold larger than the one for which we have observed mutant alleles during the screen. This screen demonstrates that mutations affecting a variety of developmental processes can be efficiently recovered from zebrafish.

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    • "Manual tracking via frame-by-frame analysis has formed the basis of contemporary knowledge and has enabled initial characterization of the larval zebrafish locomotor repertoire (Budick and O'Malley, 2000; Borla et al., 2002; McElligott and O'Malley, 2005). However, manual techniques are both laborious and limited in scope for high-throughput screens (Driever et al., 1996; Granato et al., 1996; Haffter and Nusslein- Volhard, 1996). The currently available automated tools have limitations in either refinement or time-scale. "
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    • "The crucial difference between goldfish strains and zebrafish mutants is the process of selection. Researchers have used largescale mutant screening to identify mutated loci and alleles (Mullins et al., 1994; Haffter et al., 1996; Driever et al., 1996; Amsterdam et al., 1999, 2004; Golling et al., 2002; Wienholds et al., 2003; Sivasubbu et al., 2006; Nagayoshi et al., 2008). The established zebrafish mutant strains tend to be maintained for detailed investigation of the molecular function of the responsible locus and allele (Mullins et al., 1994; Gaiano et al., 1996; vanEeden et al., 1996; Amsterdam et al., 1999). "
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    ABSTRACT: Artificial selection of post-embryonic features is known to have established morphological variation in goldfish (Carassius auratus). Although previous studies have suggested that goldfish and zebrafish are almost directly comparable at the embryonic level, little is known at the post-embryonic level. Here, we categorized the post-embryonic developmental process in the wild type goldfish into eleven different stages. We also report certain differences between the post-embryonic developmental processes of goldfish and zebrafish, especially in the skeletal systems (scales and median fin skeletons), suggesting that post-embryonic development underwent evolutionary divergence in these two teleost species. Our post-embryonic staging system of wild type goldfish paves the way for careful and appropriate comparison with other teleost species. The staging system will also facilitate comparative ontogenic analyses between wild-type and mutant goldfish strains, allowing us to closely study the relationship between artificial selection and molecular developmental mechanisms in vertebrates. This article is protected by copyright. All rights reserved. © 2015 Wiley Periodicals, Inc.
    Developmental Dynamics 08/2015; DOI:10.1002/dvdy.24340 · 2.38 Impact Factor
    • "Saturation mutagenesis screens have been performed using chemical or insertional mutagenesis [169] [170] [171] [172] and many of the mutants reproduce human genetic diseases [173] [174]. "
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    ABSTRACT: Oxidative stress is an important mechanism of chemical toxicity, contributing to developmental toxicity and teratogenesis as well as to cardiovascular and neurodegenerative diseases and diabetic embryopathy. Developing animals are especially sensitive to effects of chemicals that disrupt the balance of processes generating reactive species and oxidative stress, and those anti-oxidant defenses that protect against oxidative stress. The expression and inducibility of anti-oxidant defenses through activation of NFE2-related factor 2 (Nrf2) and related proteins is an essential process affecting the susceptibility to oxidants, but the complex interactions of Nrf2 in determining embryonic response to oxidants and oxidative stress are only beginning to be understood. The zebrafish (Danio rerio) is an established model in developmental biology and now also in developmental toxicology and redox signaling. Here we review the regulation of genes involved in protection against oxidative stress in developing vertebrates, with a focus on Nrf2 and related cap'n'collar (CNC)-basic-leucine zipper (bZIP) transcription factors. Vertebrate animals including zebrafish share Nfe2, Nrf1, Nrf2, and Nrf3 as well as a core set of genes that respond to oxidative stress, contributing to the value of zebrafish as a model system with which to investigate the mechanisms involved in regulation of redox signaling and the response to oxidative stress during embryolarval development. Moreover, studies in zebrafish have revealed nrf and keap1 gene duplications that provide an opportunity to dissect multiple functions of vertebrate NRF genes, including multiple sensing mechanisms involved in chemical-specific effects. Copyright © 2015. Published by Elsevier Inc.
    Free Radical Biology and Medicine 06/2015; DOI:10.1016/j.freeradbiomed.2015.06.022 · 5.74 Impact Factor
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