Shigenobu Shuji, Kitadate Yu, Noda Chiyo, Kobayashi Satoru
Okazaki Institute for Integrative Bioscience, National Institute for Basic Biology, National Institutes of Natural Sciences, Higashiyama, Myodaiji, Okazaki 444-8787, Japan.
Proc Natl Acad Sci U S A. 2006 Sep 12;103(37):13728-33. doi: 10.1073/pnas.0603767103. Epub 2006 Sep 1.
In many animal species, germ-line progenitors associate with gonadal somatic cells to form the embryonic gonads (EGs) that later develop into functional organ producing gametes. To explore the genetic regulation of the germ-line development, we initiated a comprehensive identification and functional analysis of the genes expressed within the EGs. First, we generated a cDNA library from gonads purified from Drosophila embryos by FACS. Using this library, we catalogued the genes expressed in the gonad by EST analysis. A total of 17,218 high-quality ESTs representing 3,051 genes were obtained, corresponding to 20% of the predicted genes in the genome. The EG transcriptome is unexpectedly distinct from that of adult gonads and includes an extremely high proportion of retrotransposon-derived transcripts. We verified 101 genes preferentially expressed in the EGs by whole-mount in situ hybridization. Within this subset, 39 and 58 genes were expressed predominantly in germ-line and somatic cells, respectively, whereas four genes were expressed in the both cell lineages. The gonad-enriched genes encompassed a variety of predicted functions. However, genes implicated in SUMOylation and protein translation, including germ-line-specific ribosomal proteins, are preferentially expressed in the germ line, whereas the expression of various retrotransposons and RNAi-related genes are more prominent in the gonadal soma. These transcriptome data are a resource for understanding the mechanism of various cellular events during germ-line development.
在许多动物物种中,生殖系祖细胞与性腺体细胞结合形成胚胎性腺(EGs),后者随后发育成产生配子的功能性器官。为了探索生殖系发育的遗传调控,我们启动了对EGs中表达基因的全面鉴定和功能分析。首先,我们通过荧光激活细胞分选(FACS)从果蝇胚胎中纯化性腺,构建了一个cDNA文库。利用这个文库,我们通过EST分析对性腺中表达的基因进行了编目。共获得了代表3051个基因的17218条高质量EST,相当于基因组中预测基因的20%。EG转录组出人意料地不同于成年性腺的转录组,并且包含极高比例的逆转座子衍生转录本。我们通过全胚胎原位杂交验证了101个在EGs中优先表达的基因。在这个子集中,分别有39个和58个基因主要在生殖系细胞和体细胞中表达,而有4个基因在这两个细胞谱系中都有表达。性腺富集基因涵盖了各种预测功能。然而,与SUMO化和蛋白质翻译相关的基因,包括生殖系特异性核糖体蛋白,在生殖系中优先表达,而各种逆转座子和RNAi相关基因在性腺体细胞中的表达更为突出。这些转录组数据是理解生殖系发育过程中各种细胞事件机制的资源。