Medaka dead end encodes a cytoplasmic protein and identifies embryonic and adult germ cells
Medaka dead end encodes a cytoplasmic protein and identifies embryonic and adult germ cells
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DOI：
10.1016/j.gep.2009.06.008
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发表时间：
2009-10-01
影响因子：
1.2
通讯作者：
Hong, Yunhan
中科院分区：
文献类型：
--
作者：
Liu, Lixiu;Hong, Ni;Hong, Yunhan
在斑马鱼中鉴定了死端（dnd）基因，该基因编码脊椎动物原始生殖细胞（PGC）发育和配子发生所必需的RNA结合蛋白。成体dnd RNA的表达仅限于非洲爪蟾的卵巢或小鼠的睾丸。其蛋白产物在鸡生殖细胞中为核蛋白，但在小鼠细胞培养物中为胞质和核蛋白。在这里，我们报告的Odnd的克隆和表达模式，（Oryzias latipes）rind基因序列比较，基因结构，连锁分析和表达表明Odnd编码青鳉Dnd同源基因。对来自五种鱼类和四足动物代表的Dnd蛋白进行系统比较，鉴定出除了RNA识别基序之外的五个先前未鉴定的保守区域。Odnd RNA由母体提供，优先与PGCs分离。在雄性和雌性中，Odnd都有表达，并且仅限于生殖细胞。在睾丸中，Odnd在精原细胞和减数分裂细胞中表达丰富，但在精子中不表达。Odnd RNA持续整个卵子发生此外，我们开发了一种双色荧光原位杂交程序，允许青鳉胚胎和成体组织中两个基因之间的表达和分布模式的精确比较。重要的是。该方法将Odnd和Ovasa共定位于睾丸生殖细胞和PGCs中。令人惊讶的是，通过细胞转染和胚胎RNA注射，我们发现ODnd在细胞培养物、卵裂胚胎和PGCs中是细胞质的。因此，青鳉dnd编码细胞质蛋白，并鉴定两种性别的胚胎和成体生殖细胞。(C)2009 Elsevier B V.保留所有权利
dead end (dnd) was identified in zebrafish as a gene encoding an RNA-binding protein essential for primordial germ cell (PGC) development and gametogenesis in vertebrates. The adult dnd RNA expression has been restricted to the ovary in Xenopus or to the testis in mouse. Its protein product is nuclear in chicken germ cells but both cytosolic and nuclear in mouse cell cultures. Here we report the cloning and expression pattern of Odnd, the medakafish (Oryzias latipes) rind gene Sequence comparison, gene structure, linkage analysis and expression demonstrate that Odnd encodes the medaka Dnd orthologue A systematic comparison of Dnd proteins from five fishes and tetrapod representatives led to the identification of five previously unidentified conserved regions besides the RNA recognition motif The Odnd RNA is maternally supplied and preferentially segregated with PGCs. Its adult expression occurs in both sexes and is restricted to germ cells In the testis, Odnd is abundant in spermatogonia and meiotic cells but absent in sperm In the ovary. Odnd RNA persists throughout oogenesis Furthermore, we developed a dual color fluorescent in situ hybridization procedure allowing for precise comparisons of expression and distribution patterns between two genes in medaka embryos and adult tissues. Importantly. this procedure co-localized Odnd and Ovasa in testicular germ cells and PGCs. Surprisingly, by cell transfection and embryo RNA injection we show that ODnd is cytoplasmic in cell cultures, cleavage embryos and PGCs Therefore, medaka dnd encodes a cytoplasmic protein and identifies embryonic and adult germ cells of both sexes. (C) 2009 Elsevier B V. All rights reserved