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首页> 外文期刊>Zoological Science >Molecular Cloning and Gene Expression of Mummichog (Fundulus heteroclitus) Runx2 During Embryogenesis
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Molecular Cloning and Gene Expression of Mummichog (Fundulus heteroclitus) Runx2 During Embryogenesis

机译：Mummichog（Fundulus heteroclitus）Runx2在胚胎发生过程中的分子克隆和基因表达

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In our previous study, we clarified the toxicity of 2,2'-dipyridyldisulfide [(PS)(2)], one of photodegradation products of a metal pyrithione that is used as an alternative antifouling paint biocides to organotin compounds in Japan. In early life stage toxicity tests, we exposed the mummichog, (Fundulus heteroclitus) to (PS)(2), and the hatched larvae subsequently displayed notochord undulations and skeletal deformities (Mochida et al., 2012). Runx2, a transcription factor of the runt family, is a key regulator in skeletal development in mammals. It is possible that (PS)(2) inhibits Runx2 gene expression, inducing the skeletal deformities in mummichog. In the present study, we cloned two Runx2 cDNAs (type I and type II) from mummichog embryos. The deduced amino acid sequences of type I and type II contain an open reading frame encoding 450 and 464 amino acid residues, respectively. The derived amino acid sequence of Fundulus Runx2 type I showed the highest identity (93.8%) with Takifugu Runx2 type I, and Fundulus Runx2 type II showed 94.6% homology with medaka Runx2. The expression level of Runx2 mRNA in the early stage series was measured using a real-time quantitative PCR assay. Expression levels tended to increase in both the blastula-gas-trula and the retinal pigmentation stage. To examine the effect of toxic compounds on skeletal formation, mummichog embryos in the late blastula to retinal pigmentation stage were exposed to (PS)(2). After exposure to (PS)(2) for one week, the expression level of Runx2 mRNA was unchanged. These results suggest that there is no inhibition of Runx2 gene expression due to (PS)(2) exposure.
机译：在我们先前的研究中，我们阐明了2,2'-联吡啶二硫化物[（PS）（2）]的毒性，这是一种金属巯氧吡啶的光降解产物，在日本用作有机锡化合物的替代防污涂料杀生物剂。在生命早期毒性测试中，我们将木乃伊（Fundulus heteroclitus）暴露于（PS）（2），并且孵出的幼虫随后显示出脊索起伏和骨骼畸形（Mochida等人，2012）。 Runx2是矮子家族的转录因子，是哺乳动物骨骼发育中的关键调节因子。 （PS）（2）可能会抑制Runx2基因表达，从而导致木乃伊的骨骼畸形。在本研究中，我们从木乃伊胚芽胚胎中克隆了两个Runx2 cDNA（I型和II型）。推导的I型和II型氨基酸序列包含一个开放阅读框，分别编码450和464个氨基酸残基。 I型Fundulus Runx2的衍生氨基酸序列与Takifugu Runx2 I型具有最高的同源性（93.8％），而Fundulus Runx2 II型与Medaka Runx2具有94.6％的同源性。使用实时定量PCR测定法检测早期系列中Runx2 mRNA的表达水平。在囊胚胃和视网膜色素沉着期中表达水平趋于增加。为了检查有毒化合物对骨骼形成的影响，将囊胚晚期至视网膜色素沉着期的木乃伊胚胎暴露于（PS）（2）。暴露于（PS）（2）1周后，Runx2 mRNA的表达水平未改变。这些结果表明，没有由于（PS）（2）暴露而导致Runx2基因表达受到抑制。

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