促性腺激素释放激素及多巴胺拮抗物地欧酮对斑节对虾卵巢组织发育的影响

摘要

This study investigated the effects of Eriocheir sinensis GnRH( EsGnRH-I ), Procambarus clarkii GnRH( PcGnRH ), domperidone( DOM)and nerve tissues on ovarian development in Penaeus monodon. The stage of ovarian and the oocyte morphology in each period were determined by paraffin sections,HE staining and fluorescence observation. The results show that P. monodon ovaries had natural autofluorescence. The fluorescence focused on the region including follicular cells,cytoplasm of developing oocyte and ma-ture oocyte which contained amount of yolk globules. The fluorescence intensity was positively correlated to ovarian developmental sta-ges(P﹤0. 01). Then we analyzed the effects of three types of hormones on the ovarian development by ovarian tissues cultured in vitro. The results showed that 50 ng·mL-1 EsGnRH-I,500 ng·mL-1 PcGnRH and 0. 3 μg·mL-1 DOM significantly increased the di-ameter of the oocytes(P ﹤0. 01),similar to the treatments of brain and thoracic nerve. Combined use of PcGnRH and DOM signifi-cantly increased the diameter of the oocytes(P ﹤0. 01). Real time quantitative PCR show that treatments of GnRH and DOM obvious-ly induced the accumulation of vitellogenin. In conclusion, crustaceans GnRH and DOM could promote the oocyte maturation of P. monodon,and combined use of them had asynergistic effect.%研究了中华绒螯蟹（ Eriocheir sinensis）促性腺激素释放激素（ EsGnRH-I）、克氏原螯虾（ Procambarus clarkii）促性腺激素释放激素（PcGnRH）、多巴胺拮抗物地欧酮（DOM）及神经组织对斑节对虾（Penaeus monodon）卵巢发育的调控作用效果。试验前使用石蜡切片、HE染色及荧光观察确定斑节对虾卵巢发育分期及各期卵母细胞形态特征。结果显示，斑节对虾卵巢具有自发荧光现象，主要集中于滤泡细胞以及富含卵黄颗粒的次级卵母细胞、成熟卵母细胞的细胞质区域，荧光强度与卵巢发育阶段具有显著正相关性（ P﹤0.01）。随后采用离体卵巢组织培养的方法研究3种激素对卵巢发育的影响。结果表明，50 ng·mL-1 EsGnRH-I、500 ng·mL-1 PcGnRH和0.3μg· mL-1 DOM 对离体卵巢的发育有明显的刺激作用，显著增大卵母细胞直径（ P﹤0.01），与脑组织、胸神经处理组效果相似；同时组合使用 PcGnRH和 DOM 能极显著促进卵母细胞发育（ P ﹤0.01）。荧光定量 PCR 结果显示PcGnRH和DOM处理后能显著增加卵黄蛋白原的积累量。说明甲壳类动物GnRH和DOM均能促进斑节对虾卵母细胞成熟，并具有协同作用。