1. College of Fisheries, Key Laboratory of Freshwater Animal Breeding, Ministry of Agriculture, Freshwater Aquaculture Collaborative Innovation Center of Hubei Province, Huazhong Agricultural University, Wuhan, 430070, China.
2. State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Science, Wuhan 430072, China.
Int J Biol Sci. 2013 Dec 1;10(1):15-24. doi: 10.7150/ijbs.7490. eCollection 2013.
C1q-like is a significant maternal factor of TNF/C1q super-family, and the abundant protein has been observed in both mature eggs of Carassius auratus and Carassius auratus gibelio, but its biological function in early embryo development has remained unclear. In this study, we firstly revealed a high level of maternal C1q-like transcript existence only in mature eggs of Carassius auratus, whereas no any maternal C1q-like transcript was observed in that of Carassius auratus gibelio. During embryonic development, the C1q-like zygotic expression begins around cardiopalmus stage in embryos of both Carassius auratus and Carassius auratus gibelio. Then, we examined the biological role of C1q-like by morpholino-mediated knockdown in early embryo development. Knockdown of CaOC1q resulted in a significant reduction of primordial germ cells (PGCs) in Carassius auratus, as shown by whole mount in situ hybridization with vasa-specific RNA probe, fluorescence immunostaining of vasa protein, and GFP imaging of the GFP-nanos1-3'UTR mRNA reporter. In vitro and in vivo evidence indicated that a microRNA, miR-430 could repress the C1q-like expression and PGC development. These data suggest that C1q-like should be a direct target of miR-430 and play an essential role in PGC development of Carassius auratus.
C1q 样蛋白是 TNF/C1q 超家族的重要母源因子，在成熟的鲤鱼和鲫鱼卵中都有丰富的表达，但它在早期胚胎发育中的生物学功能尚不清楚。在本研究中，我们首次揭示了鲤鱼成熟卵中存在高水平的母源 C1q 样转录本，而在鲫鱼成熟卵中则没有观察到任何母源 C1q 样转录本。在胚胎发育过程中，C1q 样基因在鲤鱼和鲫鱼胚胎的心脏搏动期开始有合子表达。然后，我们通过在早期胚胎发育中使用 morpholino 介导的敲低来研究 C1q 样蛋白的生物学功能。CaOC1q 的敲低导致鲤鱼原生殖细胞（PGCs）的数量显著减少，这可以通过 vasa 特异性 RNA 探针的整体原位杂交、vasa 蛋白的荧光免疫染色和 GFP-nanos1-3'UTR mRNA 报告基因的 GFP 成像来证实。体内外证据表明，一种 microRNA，miR-430 可以抑制 C1q 样蛋白的表达和 PGC 的发育。这些数据表明 C1q 样蛋白应该是 miR-430 的直接靶标，并在鲤鱼 PGC 的发育中发挥重要作用。
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