【作者】 田雪； 陈琳； 王良炎； 王磊； 马晓； 胡灿灿； 孔祥会； 聂国兴； 李学军；

【Author】 TIAN Xue;CHEN Lin;WANG Liangyan;WANG Lei;MA Xiao;HU Cancan;KONG Xianghui;NIE Guoxing;LI Xuejun;College of Fisheries, Henan Normal University;

【机构】 河南师范大学水产学院；

【摘要】 为探索硬化蛋白(Sclerostin,SOST)基因在淇河鲫(Carassius auratus in Qihe River)肌间骨骨化过程中的调控作用,本研究以淇河鲫为研究对象,利用整体骨骼染色法对肌间骨的形态发生进行观察,通过RACE方法获得SOST c DNA全长,采用q RT-PCR和western blot技术检测SOST基因在各组织及肌间骨不同骨化阶段m RNA和蛋白表达情况。结果显示,淇河鲫发育至24 dpf(day post fertilization),肌隔中出现纤维束;25 dpf髓弓小骨出现,28 dpf脉弓小骨出现,33 dpf髓弓小骨出现分叉,45 dpf肌间小骨发育完全。SOST基因在各组织中均有表达,表达水平高低依次是脑、肝、鳃、脾、肾、肠,心脏和肌肉。SOST基因在23 dpf表达量最低(P<0.05),随着肌间小骨骨化发育,SOST基因呈递增趋势,45 dpf表达量显著高于其它骨化阶段(P<0.05)。SOST m RNA和蛋白变化趋势基本相同,都在23dpf处于最低,肌间骨全部出现后表达量达到最高,与淇河鲫肌间骨的形态发生趋势一致。因此,推测SOST与淇河鲫肌间骨骨化具有一定的相关性,可调控肌间骨的分化和形成。更多还原

【Abstract】 To explore the function of gene SOST in intermuscular bone ossification of Carassius auratus in Qihe River, a modified whole bone staining method was firstly used to investigate ossification development of intermusuclar bone. Then, the q RT-PCR and western blot were conducted to detect the m RNA and protein expression of gene SOST in different ossific stages. The results showed that the ligaments were found in myoseptum at 24 dpf(day post fertilization). The earliest epineural bones were appeared at 25 dpf, and the epipleural bones were subsequently checked at 28 dpf. At 33 dpf, the epineural bones began to bifurcate. Until 45 dpf, all intermuscular bones were discovered in myoseptum. Tissue expression analysis showed that SOST m RNA exhibited extensive tissue distribution including brain, liver, spleen, gill, kidney, intestine, heart and muscle. The lowest in various ossific stages was at 23 dpf(P < 0.05). The m RNA abundance of gene SOST was increased along with the ossification development and reached a maximum level at 45 dpf(P < 0.05). The expression of gene SOST protein showed the similar trend with the m RNA and both of them varied along with the development process of intermuscular bone. Thus, we concluded that there were relationship between the gene SOST and ossification development of intermusclar bone, and the gene SOST may play an important role in morphology and ossification of intermuscular bone.更多还原