Determination in oocytes of the reproductive modes for the brine shrimp Artemia parthenogenetica
Determination in oocytes of the reproductive modes for the brine shrimp Artemia parthenogenetica
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DOI：
10.1042/bsr20090141
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发表时间：
2011-02-01
影响因子：
4
通讯作者：
Yang, Wei-Jun
中科院分区：
文献类型：
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作者：
Dai, Zhong-Min;Li, Ran;Yang, Wei-Jun
卤虫以卵生方式繁殖，产生包囊胚胎（滞育包囊），或以卵胎生方式繁殖，产生自由游动的无节幼体。环境因素，如光周期，已被应用于控制繁殖模式的贫血，但何时发生的生殖模式的确定仍然未知。分析了不同光周期条件下卵生和卵胎生卤虫卵母细胞基因表达的差异。通过消减杂交共获得692个合格的cDNA克隆，其中327个与GenBank（R）核苷酸序列数据库条目匹配。使用实时PCR在卵母细胞中分析44个cDNA（代表56个克隆）的基因表达。在这些基因中，11个（21个克隆）在进入滞育的卤虫卵母细胞中表达显著上调（P < 0.05），7个（9个克隆）表达显著下调。值得注意的是，已知的滞育相关蛋白，如ArHsp 22（卤虫热休克蛋白22）和几丁质结合蛋白被发现已经差异表达。此外，RNA干扰（RNA interference）敲低卵胎生卤虫卵母细胞中的差异表达基因polo-like kinase 1，导致产生白色胚胎而不是自由游泳的无节幼体。总之，我们的研究结果在分子水平上提供了证据，表明贫血的生殖模式已经在其生命周期的卵母细胞阶段确定。
The brine shrimp, Artemia, reproduces either oviparously, producing encysted embryos (diapause cysts), or ovoviviparously, producing free-swimming nauplii. Environmental factors, such as photoperiod, have been applied to control the reproduction mode of Anemia, but when the determination of a reproductive mode occurs remains unknown. We analysed the differential gene expression between oocytes from oviparous and ovoviviparous Artemia reared under different photoperiods. A total of 692 qualified cDNA clones were obtained by subtractive hybridization, 327 of which matched GenBank (R) Nucleotide Sequence Database entries. Gene expressions of 44 cDNAs (representing 56 clones) were analysed in oocytes using real-time PCR. Among these genes, 11 (21 clones) were significantly (P < 0.05) upregulated and 7 (9 clones) down-regulated in Artemia oocytes that subsequently enter diapause. Remarkably, known diapause-related proteins such as ArHsp22 (Artemia heat-shock protein 22) and chitin-binding proteins are found to be already differentially expressed. Furthermore, RNAi (RNA interference) knockdown of a differentially expressed gene, polo-like kinase 1, in oocyte of ovoviviparous Artemia led to the production of white embryos rather than free-swimming nauplii. In summary, our results provide evidence at the molecular level that the reproductive mode of Anemia is already determined at the oocyte stage of their life cycle.