Molecular cloning and characterization of oocyte-specific Pat1a in Rana rugosa frogs.
Molecular cloning and characterization of oocyte-specific Pat1a in Rana rugosa frogs.
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林蛙卵母细胞特异性 Pat1a 的分子克隆和表征。
DOI：
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发表时间：
2015
期刊：
影响因子：
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通讯作者：
Masahisa Nakamura
中科院分区：
文献类型：
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作者：
Yoriko Nakamura;Takehiro Iwasaki;Yosuke Umei;K. Saotome;Y. Nakajima;Shoichi Kitahara;Y. Uno;Y. Matsuda;A. Oike;Maho Kodama;Masahisa Nakamura
Pat 1基因在非洲爪蟾的未成熟卵母细胞中表达，据报道参与调节卵母细胞成熟所需的母体mRNA的翻译。然而，它仍然是未知的，当Pat 1a第一次出现在分化的两栖动物卵巢。为了解决这个问题，我们分离出全长Pat 1a的cDNA从青蛙rugosa和检查其在分化卵巢的这种青蛙的表达。在8个不同的组织检查，Pat 1a mRNA是检测到卵巢。对不同发育阶段的蝌蚪卵巢冰冻切片进行Vasa（一种生殖细胞特异性蛋白）和Pat 1a的免疫染色，发现Vasa免疫阳性信号在所有的生殖细胞中都有，而Pat 1a信号仅限于生长中的卵母细胞（直径50-200 μm），而在小的生殖细胞（直径<50 μm）中没有。40天后，注射睾酮到蝌蚪，以诱导女性到男性的性逆转，Pat 1a免疫反应性卵母细胞已完全从性逆转的性腺消失，但Vasa阳性的小生殖细胞持续存在。因此，Pat 1a将是一个很好的标记，以确定性别逆转性腺的两栖动物的性状态。此外，荧光原位杂交分析显示Pat 1a具有一个常染色体基因座，表明Pat 1a的转录可能受一种组织特异性转录因子的调控。皱纹。
The Pat1 gene is expressed in the immature oocytes of Xenopus, and is reportedly involved in regulating the translation of maternal mRNAs required for oocyte-maturation. However, it is still unknown when Pat1a first appears in the differentiating ovary of amphibians. To address this issue, we isolated the full-length Pat1a cDNA from the frog Rana rugosa and examined its expression in the differentiating ovary of this frog. Among eight different tissues examined, the Pat1a mRNA was detectable in only the ovary. When frozen sections from the ovaries of tadpoles at various stages of development were immunostained for Vasa-a germ cell-specific protein-and Pat1a, Vasa-immunopositive signals were observed in all of the germ cells, whereas Pat1a signals were confined to the growing oocytes (50-200 μm in diameter), and absent from small germ cells (<50 μm in diameter). Forty days after testosterone injection into tadpoles to induce female-to-male sex-reversal, Pat1a-immunoreactive oocytes had disappeared completely from the sex-reversed gonad, but Vasa-positive small germ cells persisted. Thus, Pat1a would be a good marker for identifying the sexual status of the sex-reversing gonad in amphibians. In addition, fluorescence in situ hybridization analysis showed Pat1a to have an autosomal locus, suggesting that Pat1a transcription is probably regulated by a tissue-specific transcription factor in R. rugosa.