大头金蝇卵不同发育时间形态变化及基因表达差异研究

发布时间：2019-02-17 12:11

【摘要】：目的研究大头金蝇(Chrysomya megacephala)卵不同发育时间的形态变化和基因表达差异规律。方法从大头金蝇成虫产完卵后,取出卵块,记时为0 h,每隔2 h取卵10枚,直至有幼虫孵出,分别用体视显微镜和扫描电镜观察其不同发育时间的形态学变化。提取0、2、4、6和8 h等5个发育时间点的蝇卵RNA,应用RT-PCR比较循环阈值(threshold cycle,CT值)法测定bicoid、slalom和几丁质合成酶(chitin synthase)基因相对表达水平,SPSS19.0统计学软件对CT值及时间进行拟合直线方程、等比级数曲线方程和指数方程分析。结果体视显微镜下观察,大头金蝇卵产后0~4h形态学变化不明显,6h有体节形成,8 h开始出现皱缩,9 h有幼虫孵出;扫描电镜下观察,卵产后0~4 h时变化不明显,卵孔一端略向外突出,卵孔周围光滑,6 h后卵孔一端向内凹陷,卵孔周围有不规则突起形成,8 h时凹陷明显,9 h已发育为幼虫。大头金蝇卵在不同发育时间bicoid、slalom和chitin synthase等3个基因的相对表达水平随着时间变化呈一定的规律变化,相关回归方程式3个基因CT值的差异均有统计学意义(P0.05)。结论体视显微镜和扫描电镜下大头金蝇卵形态结构随时间改变而发生变化。大头金蝇卵不同发育时间的bicoid、slalom和chitin synthase基因表达水平有差异。
[Abstract]:Objective to study the morphological changes and gene expression of (Chrysomya megacephala) eggs in Drosophila macrocephala. Methods the eggs were taken out from adults of Drosophila macrocephala at 0 h and 10 eggs were taken out every 2 hours until the larvae hatched. The morphological changes of the eggs at different developmental times were observed by stereomicroscopy and scanning electron microscope. The relative expression levels of bicoid,slalom and chitin synthase (chitin synthase) gene were determined by RT-PCR comparative circulation threshold (threshold cycle,CT) method. The CT value and time were fitted by SPSS19.0 software, and the linear equation, the equal-ratio series curve equation and the exponential equation were analyzed. Results under stereoscopic microscope, the morphological changes were not obvious at 0h, 6h, 8h, and 9h, respectively. Under scanning electron microscope, the changes were not obvious at 0h postpartum, the one end of egg foramina was slightly protruded out, and the circumference of egg foramen was smooth. After 6 h, one end of egg foramen was concave inward, irregular protuberance formed around egg foramen, and obvious depression was observed at 8 h. The larva has developed into larva in 9 h. The relative expression level of three genes (bicoid,slalom and chitin synthase) in the egg of Drosophila macrocephala changed regularly with time, and the CT values of the three genes in the regression equation were significantly different (P0.05). Conclusion the morphological structure of Drosophila macrocephala eggs changed with time under stereoscopic microscope and scanning electron microscope. The expression levels of bicoid,slalom and chitin synthase genes in Drosophila macrocephala eggs at different developmental times were different.
【作者单位】：贵州省贵阳医学院法医学系/法医司法鉴定中心;贵州省安顺市公安局刑侦支队;
【基金】：2013年贵州省高校工程技术研究中心项目(黔科合KY字[2013]114号) 2012年贵州省教育厅产学研基地项目(黔教合KY字[2012]031号) 贵州省卫生厅科学技术基金项目(WT2013-1) 贵阳医学院院基金项目(院基金合同字第021号)~~
【分类号】：D919

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