2015, 36(5): 1-7. doi: 10.11758/yykxjz.20150501
关键词： 半滑舌鳎 , Runx2基因 , Osterix基因 , mRNA表达 , 早期发育阶段
Keywords： Cynoglossus semilaevis , Runx2 , Osterix , mRNA expression , Early developmental stages
转录因子Runx2和Osterix在高等脊椎动物骨骼发育的调控过程中具有重要作用。本研究以Runx2和Osterix基因作为研究对象,利用实时荧光定量PCR技术分析了两个转录因子在半滑舌鳎(Cynoglossus semilaevis)成鱼的组织表达分布及其在早期发育阶段的表达水平变化。研究结果显示,Runx2和Osterix在半滑舌鳎成鱼的13个组织中具有较广泛的分布,并在脾组织中表达量最高,在背鳍、脑等组织中次之,在肝、肠、胃等组织中仅有微弱的表达。在半滑舌鳎早期发育阶段(卵期、仔鱼、稚鱼和幼鱼),Runx2和Osterix的时序性表达特征如下:1) 孵化前,Runx2在卵裂期与原肠期的表达水平最高,在胚胎期最低,而Osterix表达水平呈逐渐升高的趋势并在胚胎期达到最大值,表明两个基因功能作用的起始可能存在一定的时序性;2) Runx2和Osterix在1-5日龄仔鱼阶段的表达水平均呈现先升高后降低的趋势,而在10-90日龄稚、幼鱼阶段未呈现特定的变动规律,表达量在一定范围内呈波动变化;3) 相关性分析结果显示,两个基因的表达量在孵化后(1-90日龄)存在显著的相关性。研究结果表明,Runx2和Osterix基因可能共同参与半滑舌鳎早期发育阶段生长发育调控,但其与骨骼发生、发育的相关性有待进一步研究。
Transcription factors Runx2 and Osterix play a vital role in the formation and differentiation of the skeleton in mammals. To date, there have been few reports on the regulatory mechanisms of the skeleton development in fish. In this study, we cloned the cDNA sequences of Runx2 and Osterix in Cynoglossus semilaevis, and examined their expression patterns in 13 tissues of both female and male adults using qRT-PCR. The results showed that Runx2 and Osterix mRNA were highly expressed in spleen, and moderately expressed in dorsal fin and brain. Next we detected and measured the levels of Runx2 and Osterix mRNA at early developmental stages including the egg, larva, juvenile and fingerling stages. The temporal expression patterns are: 1) the mRNA level of Osterix was always significantly higher than Runx2 during all developmental stages in this study; 2) at stages prior to hatching, the highest level of Runx2 was observed in the cleavage and gastrula stages and then it dropped to a very low level, and the Osterix mRNA level increased and reached the peak in the embryonic stage; 3) at early larvae stages(1-5 days old), the expression of both Runx2 and Osterix first increased followed by a decline; 4) as for the 10-90 day-old fish, there was no variation in the expression pattern of Runx2 or Osterix mRNA. These results suggested that Runx2 and Osterix could participate in the regulation of early development in C. semilaevis. However, further studies are needed to explore their roles in the skeleton development.
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