Transgenic medaka enables easy oocytes detection in live fish

Kinoshita, Masato; Okamoto, Gensei; Hirata, Takashi; Shinomiya, Ai; Kobayashi, Tohru; Kubo, Yuko; Hori, Hiroshi; Kanamori, Akira

doi:10.1002/mrd.20942

Cited by 25 publications

(26 citation statements)

References 14 publications

(12 reference statements)

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“…Gene expression of an ovary-specific marker, 42Sp50, 20 was examined in non-irradiated and irradiated testes of wt Hd-rR and p53-deficient medaka males 7 days after 5 Gy γ -ray irradiation. As clearly shown in Figure 8a, 42Sp50 transcripts were amplified in non-irradiated and irradiated p53-deficient medaka testes, as well as in ovaries of wt and p53-deficient medaka.…”

Section: Resultsmentioning

confidence: 99%

Gamma-ray irradiation promotes premature meiosis of spontaneously differentiating testis–ova in the testis of p53-deficient medaka (Oryzias latipes)

Yasuda

Oda

et al. 2012

Cell Death Dis

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In this study, the roles of p53 in impaired spermatogenic male germ cells of p53-deficient medaka were investigated by analyzing histological changes, and gene expressions of 42Sp50, Oct 4 and vitellogenin (VTG2) by RT-PCR or in situ hybridization in the testes. We found that a small number of oocyte-like cells (testis–ova) differentiated spontaneously in the cysts of type A and early type B spermatogonia in the p53-deficient testes, in contrast to the wild-type (wt) testes in which testis–ova were never found. Furthermore, ionizing radiation (IR) irradiation increased the number of testis–ova in p53-deficient testes, increased testis–ova size and proceeded up to the zygotene or pachytene stages of premature meiosis within 14 days after irradiation. However, 28 days after irradiation, almost all the testis–ova were eliminated presumably by p53-independent apoptosis, and spermatogenesis was restored completely. In the wt testis, IR never induced testis–ova differentiation. This is the first study to demonstrate the pivotal role of the p53 gene in the elimination of spontaneous testis–ova in testes, and that p53 is not indispensable for the restoration of spermatogenesis in the impaired testes in which cell cycle regulation is disturbed by IR irradiation.

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Section: Resultsmentioning

confidence: 99%

Gamma-ray irradiation promotes premature meiosis of spontaneously differentiating testis–ova in the testis of p53-deficient medaka (Oryzias latipes)

Yasuda

Oda

et al. 2012

Cell Death Dis

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“…42sp43 is a nine zinc-finger protein responsible for 5S rRNA binding, but unlike TFIIIA it is not able to bind the gene and it is not implicated in its transcriptional regulation [133]. The only reference in fish is about Oryzias latipes where both 42sp50 and 42sp43 gene transcripts were found to be ovary specific [141,142]. A transgenic medaka line constructed using a relative short upstream sequence and 3’UTR from the 42sp50 gene allowed oocytes to be fluorescently labeled; fluorescence beginning as early as five days after hatch [142].…”

Section: Novel Intersex Markers In Mulletsmentioning

confidence: 99%

“…The only reference in fish is about Oryzias latipes where both 42sp50 and 42sp43 gene transcripts were found to be ovary specific [141,142]. A transgenic medaka line constructed using a relative short upstream sequence and 3’UTR from the 42sp50 gene allowed oocytes to be fluorescently labeled; fluorescence beginning as early as five days after hatch [142]. Oocytes artificially induced in the testes of medaka were also fluorescently labeled [142].…”

Section: Novel Intersex Markers In Mulletsmentioning

confidence: 99%

“…A comparison of the germ cell specific promoters reveals that they all possess a small size (100 bp). This is exemplified by the transgenic O. latipes line [142] with fluorescent ovaries produced using the short promoter sequence of 42sp50 . It is not clear how such short promoters are able to specify germ cell activation and somatic silencing but they suggest that enhancer-dependent interactions are not required.…”

Section: Novel Intersex Markers In Mulletsmentioning

confidence: 99%

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Mugilid Fish Are Sentinels of Exposure to Endocrine Disrupting Compounds in Coastal and Estuarine Environments

et al. 2014

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Effects on fish reproduction can result from a variety of toxicity mechanisms first operating at the molecular level. Notably, the presence in the environment of some compounds termed endocrine disrupting chemicals (EDCs) can cause adverse effects on reproduction by interfering with the endocrine system. In some cases, exposure to EDCs leads to the animal feminization and male fish may develop oocytes in testis (intersex condition). Mugilid fish are well suited sentinel organisms to study the effects of reproductive EDCs in the monitoring of estuarine/marine environments. Up-regulation of aromatases and vitellogenins in males and juveniles and the presence of intersex individuals have been described in a wide array of mullet species worldwide. There is a need to develop new molecular markers to identify early feminization responses and intersex condition in fish populations, studying mechanisms that regulate gonad differentiation under exposure to xenoestrogens. Interestingly, an electrophoresis of gonad RNA, shows a strong expression of 5S rRNA in oocytes, indicating the potential of 5S rRNA and its regulating proteins to become useful molecular makers of oocyte presence in testis. Therefore, the use of these oocyte markers to sex and identify intersex mullets could constitute powerful molecular biomarkers to assess xenoestrogenicity in field conditions.

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“…42Sp50 is abundantly expressed in oocytes (Kinoshita et al 2009). In these studies, induction of testis ova occurred for a 2-week exposure to 830 ng estradiol-benzoate/L.…”

Section: Endocrine Disrupting Chemicalsmentioning

confidence: 98%

Transgenic fish systems and their application in ecotoxicology

Lee

Green

Tyler

2014

Critical Reviews in Toxicology

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The use of transgenics in fish is a relatively recent development for advancing understanding of genetic mechanisms and developmental processes, improving aquaculture, and for pharmaceutical discovery. Transgenic fish have also been applied in ecotoxicology where they have the potential to provide more advanced and integrated systems for assessing health impacts of chemicals. The zebrafish (Daniorerio) is the most popular fish for transgenic models, for reasons including their high fecundity, transparency of their embryos, rapid organogenesis and availability of extensive genetic resources. The most commonly used technique for producing transgenic zebrafish is via microinjection of transgenes into fertilized eggs. Transposon and meganuclease have become the most reliable methods for insertion of the genetic construct in the production of stable transgenic fish lines. The GAL4-UAS system, where GAL4 is placed under the control of a desired promoter and UAS is fused with a fluorescent marker, has greatly enhanced model development for studies in ecotoxicology. Transgenic fish have been developed to study for the effects of heavy metal toxicity (via heat-shock protein genes), oxidative stress (via an electrophile-responsive element), for various organic chemicals acting through the aryl hydrocarbon receptor, thyroid and glucocorticoid response pathways, and estrogenicity. These models vary in their sensitivity with only very few able to detect responses for environmentally relevant exposures. Nevertheless, the potential of these systems for analyses of chemical effects in real time and across multiple targets in intact organisms is considerable. Here we illustrate the techniques used for generating transgenic zebrafish and assess progress in the development and application of transgenic fish (principally zebrafish) for studies in environmental toxicology. We further provide a viewpoint on future development opportunities.

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Transgenic medaka enables easy oocytes detection in live fish

Cited by 25 publications

References 14 publications

Gamma-ray irradiation promotes premature meiosis of spontaneously differentiating testis–ova in the testis of p53-deficient medaka (Oryzias latipes)

Gamma-ray irradiation promotes premature meiosis of spontaneously differentiating testis–ova in the testis of p53-deficient medaka (Oryzias latipes)

Mugilid Fish Are Sentinels of Exposure to Endocrine Disrupting Compounds in Coastal and Estuarine Environments

Transgenic fish systems and their application in ecotoxicology

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