Phenotypic analysis of a novel <i>chordin</i> mutant in medaka

Takashima, Shigeo; Shimada, Atsuko; Kobayashi, Daigo; Yokoi, Hayato; Narita, Takanori; Jindo, Tomoko; Kage, Tomofumi; Kitagawa, Tadao; Kimura, Tetsuaki; Sekimizu, Koshin; Miyake, Akimitsu; Setiamarga, Davin H. E.; Murakami, Ryutaro; Tsuda, Sachiko; Ooki, Shinya; Kakihara, Ken; Hojo, Motoki; Naruse, Kiyoshi; Mikami, Hiroshi; Shima, Akihiro; Ishikawa, Yuji; Araki, Kazuo; Saga, Yumiko; Takeda, Hiroyuki

doi:10.1002/dvdy.21245

Cited by 31 publications

(36 citation statements)

References 54 publications

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“…A digoxigenin (DIG)-labeled RNA probe was generated using the DIG RNA labeling kit (Roche). WISH was performed as described previously (Takashima et al 2007). …”

Section: Methodsmentioning

confidence: 99%

pnp4aIs the Causal Gene of the Medaka Iridophore Mutantguanineless

Kimura

Takehana

Naruse

2017

G3 Genes|Genomes|Genetics

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See-through medaka lines are suitable for observing internal organs throughout life. They were bred by crossing multiple color mutants. However, some of the causal genes for these mutants have not been identified. The medaka has four pigment cell types: black melanophores, yellow xanthophores, white leucophores, and silvery iridophores. The causal genes of melanophore, xanthophore, and leucophore mutants have been elucidated, but the causal gene for the iridophore mutant remains unknown. Here, we describe the iridophore mutant, guanineless (gu), which exhibits a strong reduction in visible iridophores throughout its larval to adult stages. The gu locus was previously mapped to chromosome 5, but was located near the telomeric region, making it difficult to integrate into the chromosome. We sought the causal gene of gu using synteny analysis with the zebrafish genome and found a strong candidate, purine nucleoside phosphorylase 4a (pnp4a). Gene targeting and complementation testing showed that pnp4a is the causal gene of gu. This result will allow the establishment of inbred medaka strains or other useful strains with see-through phenotypes without major disruption in the genetic background of each strain.KEYWORDS medaka iridophore pnp4a pigment mutant guanineless Medaka embryos are transparent and develop externally, making them suitable for the observation of internal organ developmental processes. However, because of the gradual appearance of pigment cells, observation of internal organs from the early juvenile to adult stages becomes difficult. To solve this problem, the see-through strains ST-II, ST-V, and SK2 were bred by crossing multiple medaka color mutants (Wakamatsu et al. 2001;Ohshima et al. 2013). In zebrafish, the casper fish were made for this purpose. The casper fish are doubly mutant for nacre (microphthalmia-associated transcription factor a mutant) and roy orbison. They show a complete lack of melanophores and iridophores and have an almost transparent body from the early juvenile to adult stage (White et al.

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“…A digoxigenin (DIG)-labeled RNA probe was generated using the DIG RNA labeling kit (Roche). WISH was performed as described previously (Takashima et al 2007). …”

Section: Methodsmentioning

confidence: 99%

pnp4aIs the Causal Gene of the Medaka Iridophore Mutantguanineless

Kimura

Takehana

Naruse

2017

G3 Genes|Genomes|Genetics

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“…Whole-mount in situ hybridization was performed as previously described (Takashima et al, 2007). For tbx2 (ENSORLG00000014792 and ENSORLG00000010011) primers for DIG-labeled RNA probes are listed in supplementary material Table S10.…”

Section: Whole-mount In Situ Hybridizationmentioning

confidence: 99%

Large hypomethylated domains serve as strong repressive machinery for key developmental genes in vertebrates

et al. 2014

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DNA methylation is a fundamental epigenetic modification in vertebrate genomes and a small fraction of genomic regions is hypomethylated. Previous studies have implicated hypomethylated regions in gene regulation, but their functions in vertebrate development remain elusive. To address this issue, we generated epigenomic profiles that include base-resolution DNA methylomes and histone modification maps from both pluripotent cells and mature organs of medaka fish and compared the profiles with those of human ES cells. We found that a subset of hypomethylated domains harbor H3K27me3 (K27HMDs) and their size positively correlates with the accumulation of H3K27me3. Large K27HMDs are conserved between medaka and human pluripotent cells and predominantly contain promoters of developmental transcription factor genes. These key genes were found to be under strong transcriptional repression, when compared with other developmental genes with smaller K27HMDs. Furthermore, human-specific K27HMDs show an enrichment of neuronal activityrelated genes, which suggests a distinct regulation of these genes in medaka and human. In mature organs, some of the large HMDs become shortened by elevated DNA methylation and associate with sustained gene expression. This study highlights the significance of domain size in epigenetic gene regulation. We propose that large K27HMDs play a crucial role in pluripotent cells by strictly repressing key developmental genes, whereas their shortening consolidates long-term gene expression in adult differentiated cells.

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“…Whole-mount in situ hybridization was performed as described previously (Takashima et al, 2007). Signals were visualized with NBT/BCIP tablets (Roche), BM Purple (Roche) or Fast Red tablets (Roche).…”

Section: Whole-mount In Situ Hybridizationmentioning

confidence: 99%

Modular development of the teleost trunk along the dorsoventral axis and zic1/zic4 as selector genes in the dorsal module

et al. 2013

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SUMMARYTeleost fish exhibit remarkable diversity in morphology, such as fins and coloration, particularly on the dorsal side. These structures are evolutionary adaptive because their back is highly visible to other individuals. However, owing to the late phenotypic appearance (from larva to adult) and lack of appropriate mutants, the genetic mechanisms that regulate these dorsoventrally asymmetric external patterns are largely unknown. To address this, we have analyzed the spontaneous medaka mutant Double anal fin (Da), which exhibits a mirror-image duplication of the ventral half across the lateral midline from larva to adult. Da is an enhancer mutant for zic1 and zic4 in which their expression in dorsal somites is lost. We show that the dorsoventral polarity in Da somites is lost and then demonstrate using transplantation techniques that somites and their derived tissues globally determine the multiple dorsal-specific characteristics of the body (fin morphology and pigmentation) from embryo to adult. Intriguingly, the zic1/zic4 expression in the wild type persists throughout life in the dorsal parts of somite derivatives, i.e. the myotome, dermis and vertebrae, forming a broad dorsal domain in the trunk. Comparative analysis further implies a central role for zic1/zic4 in morphological diversification of the teleost body. Taken together, we propose that the teleost trunk consists of dorsal/ventral developmental modules and that zic1/zic4 in somites function as selector genes in the dorsal module to regulate multiple dorsal morphologies.

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Phenotypic analysis of a novel chordin mutant in medaka

Cited by 31 publications

References 54 publications

pnp4aIs the Causal Gene of the Medaka Iridophore Mutantguanineless

pnp4aIs the Causal Gene of the Medaka Iridophore Mutantguanineless

Large hypomethylated domains serve as strong repressive machinery for key developmental genes in vertebrates

Modular development of the teleost trunk along the dorsoventral axis and zic1/zic4 as selector genes in the dorsal module

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