Temporal and Spatial Differential Expression of Chicken Germline-specific Proteins cDAZL, CDH and CVH During Gametogenesis
Temporal and Spatial Differential Expression of Chicken Germline-specific Proteins cDAZL, CDH and CVH During Gametogenesis
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DOI：
10.1262/jrd.09-218a
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发表时间：
2010-06-01
影响因子：
1.8
通讯作者：
Hattori, Masa-aki
中科院分区：
文献类型：
--
作者：
Kito, Gakushi;Aramaki, Shinya;Hattori, Masa-aki
Dazl基因编码的DAZL蛋白是脊椎动物配子发生所必需的生殖细胞特异性RNA结合蛋白，鸡的Dazl基因也已在原始生殖细胞中被发现。然而，鸡DAZL（cDAZL）的时空表达及其在生殖细胞发育中的分子作用仍然是个谜。在这里，我们研究了亚细胞分布和表达的cDAZL在各个阶段，通过使用针对其C-末端区域的多克隆抗体，并将它们与额外的生殖系特异性蛋白鸡血管同源物（CVH）和鸡死端同源物（CDH）进行比较。Western blot分析显示，cDAZL在胚胎性腺和未成熟鸡睾丸中有一条带，而在未成熟鸡卵巢中没有检测到带。荧光免疫组化显示cDAZL存在于循环PGCs的细胞核和细胞质中。cDAZL和CVH阳性细胞共存于胚胎性腺和未成熟鸡睾丸中，分布于曲细精管基膜附近。有趣的是，未成熟鸡卵巢中未发现cDAZL，而CVH和CDH存在于生殖细胞中。总的来说，三种生殖系特异性蛋白质在鸡生殖细胞中表达，但它们的表达模式在时间和空间上是不同的。
The Deleted in Azoospermia-Like (DAZL) protein coded by Dazl gene is a germline-specific RNA-binding protein essential for gametogenesis in vertebrates, and the chicken Dazl gene has also been identified in primordial germ cells (PGCs). However, the temporal and spatial expression of chicken DAZL (cDAZL) and its molecular role in germ cell development remain enigmatic. Here, we investigated the subcellular distribution and expression of cDAZL at the various stages by using a polyclonal antibody raised against its C-terminal region and compared them with those of additional germline-specific proteins chicken vasa homologue (CVH) and chicken dead end homologue (CDH). Western blot analysis for cDAZL revealed a single band in the embryonic gonads and premature chicken testis, whereas no band was detected in the premature chicken ovary. Fluorescent immunohistochemistry revealed that cDAZL was present in the nucleus and cytoplasm of circulating PGCs. Cells positive for cDAZL and CVH coexisted in the embryonic gonads and premature chicken testis, in which they were distributed near the basement membrane of seminiferous tubules. Of interest, cDAZL was not found in the premature chicken ovary, whereas CVH and CDH were present in germ cells. Collectively, three germline-specific proteins are expressed in chicken germ cells, but their patterns of expression are temporally and spatially distinct.