褐牙鲆作为一种重要的经济鱼种,有一个特殊的变态发育时期,其间骨骼形态和位置都发生很大改变。本研究利用三重染色技术,对褐牙鲆变态发育时期不同阶段的仔鱼整体骨骼进行染色,并对各阶段仔鱼及部分成鱼组织,采用特异性引物对骨骼发育的相关基因Bmp2,Bmp4和Sox9基因片段进行了克隆和半定量分析。研究表明,随着仔鱼变态发育,部分软骨逐渐转化为硬骨,头部筛骨呈不对称分化,额骨和顶骨位置发生改变,口形态改变,鳃丝软骨成骨,脊椎逐渐发育成熟,随之侧线位置发生改变。Sox9以及Bmp2、Bmp4基因在这时期中存在着差异表达,对骨骼发育有一定的调控作用。成鱼中,Bmp2和Sox9只在鳃、肠道和肝脏中有表达,且鳃中表达量较高。Sox9出膜后表达量突然增加,头部表达量较高,变态发育中开始递减,在变态高峰期明显减少;而Bmp4表达量则是出膜后明显减少,随后在变态高峰期表达量有增加,总体趋势表达量减少。 Paralichthys olivaceus, as an important economic species, has a special period of metamorphosis, during which the shape and location of the bones are greatly changed. In this study, the triplex staining technique was used to stain the whole bones of larvae at different stages during the metamorphosis stage of the Japanese flounder, and the Bmp2, Bmp4 and Sox9 genes related to bone development Fragments were cloned and semi-quantitatively analyzed. Studies have shown that with the larval metamorphosis, some of the cartilage gradually transformed into hard bones, the head of the ethmoid bone showed asymmetric differentiation, the frontal bone and the parietal bone changed, the shape of the mouth changed, the gill filamentous cartilage became osteogenic and the spine gradually developed and matured. Lateral position changes. Sox9 and Bmp2, Bmp4 genes in this period there are differences in the expression of bone development has a certain regulatory role. Into the fish, Bmp2 and Sox9 only in the gill, intestinal and liver expression, and the gill expression higher. Sox9 expression suddenly increased after the membrane, the head of high expression, metamorphosis began to decline, decreased significantly in the abnormal metamorphosis; while the expression of Bmp4 was significantly reduced after the membrane, and then increased in the metamorphosis peaked , The overall trend of expression decreased.