Gabrieli Paolo, Falaguerra Andrea, Siciliano Paolo, Gomulski Ludvik M, Scolari Francesca, Zacharopoulou Antigone, Franz Gerald, Malacrida Anna R, Gasperi Giuliano
Department of Animal Biology, University of Pavia, Piazza Botta 9, 27100 Pavia, Italy.
BMC Dev Biol. 2010 Jan 26;10:12. doi: 10.1186/1471-213X-10-12.
In embryos the maternal-to-zygotic transition (MTZ) integrates post-transcriptional regulation of maternal transcripts with transcriptional activation of the zygotic genome. Although the molecular mechanisms underlying this event are being clarified in Drosophila melanogaster, little is know about the embryogenic processes in other insect species. The recent publication of expressed sequence tags (ESTs) from embryos of the global pest species Ceratitis capitata (medfly) has enabled the investigation of embryogenesis in this species and has allowed a comparison of the embryogenic processes in these two related dipteran species, C. capitata and D. melanogaster, that shared a common ancestor 80-100 mya.
Using a novel PCR-based sexing method, which takes advantage of a putative LTR retrotransposon MITE insertion on the medfly Y chromosome, the transcriptomes of individual early male and female embryos were analysed using RT-PCR. This study is focused on two crucial aspects of the onset of embryonic development: sex determination and cellular blastoderm formation. Together with the three known medfly genes (Cctransformer, Cctransformer2 and Ccdoublesex), the expression patterns of other medfly genes that are similar to the D. melanogaster sex-determination genes (sisterlessA, groucho, deadpan, Sex-lethal, female lethal d, sans fille and intersex) and four cellular blastoderm formation genes (Rho1, spaghetti squash, slow-as-molasses and serendipity-alpha) were analyzed, allowing us to sketch a preliminary outline of the embryonic process in the medfly. Furthermore, a putative homologue of the Zelda gene has been considered, which in D. melanogaster encodes a DNA-binding factor responsible for the maternal-to-zygotic transition.
Our novel sexing method facilitates the study of i) when the MTZ transition occurs in males and females of C. capitata, ii) when and how the maternal information of "female-development" is reprogrammed in the embryos and iii) similarities and differences in the regulation of gene expression in C. capitata and D. melanogaster. We suggest a new model for the onset of the sex determination cascade in the medfly: the maternally inherited Cctra transcripts in the female embryos are insufficient to produce enough active protein to inhibit the male mode of Cctra splicing. The slow rate of development and the inefficiency of the splicing mechanism in the pre-cellular blastoderm facilitates the male-determining factor (M) activity, which probably acts by inhibiting CcTRA protein activity.
在胚胎中,母源 - 合子转变(MTZ)将母源转录本的转录后调控与合子基因组的转录激活整合在一起。尽管黑腹果蝇中这一事件的分子机制正在被阐明,但对于其他昆虫物种的胚胎发育过程却知之甚少。全球害虫地中海实蝇胚胎的表达序列标签(EST)的最新发表,使得对该物种胚胎发育的研究成为可能,并能对这两个在80 - 100百万年前拥有共同祖先的相关双翅目物种——地中海实蝇和黑腹果蝇的胚胎发育过程进行比较。
利用一种基于PCR的新型性别鉴定方法,该方法利用了地中海实蝇Y染色体上一个假定的LTR反转录转座子MITE插入,通过RT - PCR分析了单个早期雄性和雌性胚胎的转录组。本研究聚焦于胚胎发育起始的两个关键方面:性别决定和细胞胚盘形成。连同三个已知的地中海实蝇基因(Cctransformer、Cctransformer2和Ccdoublesex),分析了其他与黑腹果蝇性别决定基因(sisterlessA、groucho、deadpan、Sex - lethal、female lethal d、sans fille和intersex)相似的地中海实蝇基因以及四个细胞胚盘形成基因(Rho1、spaghetti squash、slow - as - molasses和serendipity - alpha)的表达模式,这使我们能够勾勒出地中海实蝇胚胎发育过程的初步轮廓。此外,还考虑了Zelda基因的一个假定同源物,在黑腹果蝇中该基因编码一种负责母源 - 合子转变的DNA结合因子。
我们的新型性别鉴定方法有助于研究:i)地中海实蝇的雄性和雌性中MTZ转变何时发生;ii)胚胎中“雌性发育”的母源信息何时以及如何被重新编程;iii)地中海实蝇和黑腹果蝇基因表达调控的异同。我们提出了一个地中海实蝇性别决定级联起始的新模型:雌性胚胎中母源遗传的Cctra转录本不足以产生足够的活性蛋白来抑制Cctra的雄性剪接模式。在前细胞胚盘阶段发育速度缓慢和剪接机制效率低下有利于雄性决定因子(M)的活性,其可能通过抑制CcTRA蛋白活性起作用。