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Searching cytoplasmic factors involved in sex determination of vertebrate

寻找参与脊椎动物性别决定的细胞质因子

基本信息

批准号：
16570188
负责人：
MIURA Ikuo
金额：
$ 2.24万
依托单位：
HIROSHIMA UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16570188/
关键词：
Sex determination Cytoplasmic factor Vertebrate Rana rugosa

项目摘要

Sex is a major mechanism to be essentially involved in the production of biological diversities and evolution in the organisms. So far, researches have been focused on a master sex determining gene on sex chromosomes. Recently, we have got some data that support existence of cytoplasmic factors in eggs involved in sex determination of frogs. The present study focused on the following three points.1.WolbachiaTo identify Wolbachia bacteria in the eggs of frog, Rana rugosa, we amplified a gene specific to the FtsZ gene of Wolbachia from genomic DNA and cDNA of the frog eggs. As the results, amplified products were got from eggs of Isehara and Hiroshima populations. However, their nucleotide sequences did not show any similalities to the FtsZ gene.2.MitochondriaTo identify any new genes involved in sex determination within the mitochondiral genome, we determined mitochondrial genome organization of the two local populations, Isehara and Hiroshima, because their hybrids show striking bias of sex ratio according to the cytoplasm. As the results, we found one unique sequence within the D-loop region of the Isehara population. That can encode 60-100 amino acids, comprising of repeats of ten amino acids, from every three frame of both strands. The ten amino acids are similar to unknown protein of the protozoan pathogen Trypanosoma cruzi.3.Maternal factorUsing RNA subtraction between WW and ZZ eggs and differential display between WW and ZZ embryos, we isolated some genes highly expressed only in each of the two kinds of eggs or embryos. In WW eggs, one galectin and mitochondrial cox I were highly expressed, while in ZZ eggs, homeodomain dbx I and protein kinase LYK-5 were highly expressed. From the WW embryos, we isolated a new W-linked glycoprotein gene, which was expressed specifically in ZW female gonads during sex determinination.

性是生物多样性产生和生物进化的主要机制。到目前为止，研究主要集中在性染色体上的主要性别决定基因上。近年来，我们获得了一些数据，支持蛙卵中存在参与性别决定的细胞质因子。本研究主要围绕以下三点展开。为了鉴定蛙蛙卵中的沃尔巴克氏菌，我们从蛙卵的基因组DNA和cDNA中扩增出沃尔巴克氏菌FtsZ基因的特异性基因。结果，从石原和广岛种群的卵中获得了扩增产物。然而，它们的核苷酸序列与FtsZ基因没有任何相似之处。为了在线粒体基因组中发现任何与性别决定有关的新基因，我们对两个当地种群Isehara和Hiroshima的线粒体基因组组织进行了测定，因为它们的杂交后代显示出明显的细胞质性别比例偏差。结果，我们在Isehara种群的D-loop区域发现了一个独特的序列。它可以编码60-100个氨基酸，包括10个氨基酸的重复，从两条链的每三个框架中。这10个氨基酸与原生动物病原体克氏锥虫的未知蛋白相似。利用WW和ZZ卵间的RNA减法和WW和ZZ胚间的差异展示，分离出仅在两种卵或胚中高表达的部分基因。WW卵高表达1种凝集素和线粒体cox I， ZZ卵高表达同源结构域dbx I和蛋白激酶LYK-5。我们从WW胚胎中分离到一个新的w -连锁糖蛋白基因，该基因在性别决定过程中在ZW雌性性腺中特异性表达。