Cre重组酶在软骨组织特异性转基因小鼠中表达的时空分布  

Expression and distribution of Cre recombinase in chondrocyte-specific Cre transgenic mice

作　　者：张继帅[1] 谭晓红[1] 王友亮[1] 程萱[1] 高翔[2] 杨晓[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100071 [2]南京大学模式生物研究中心,南京210093

出　　处：《军事医学科学院院刊》2004年第6期509-512,共4页Bulletin of the Academy of Military Medical Sciences

基　　金：国家杰出青年基金 (3 0 0 2 5 0 2 8);国家"863"计划(2 0 0 1AA2 160 81);国家攻关项目 (2 0 0 1BA710B)

摘　　要：目的 :检测Cre重组酶在软骨组织特异性Cre重组酶转基因小鼠 (Col2al Cre)表达的时空分布。方法 :将Col2a1 Cre转基因小鼠和ROSA2 6报告小鼠杂交 ,得到的双转基因小鼠中表达Cre重组酶的部位将表达LacZ基因。通过LacZ染色可直接观察不同发育阶段转基因小鼠中Cre重组酶表达的组织特异性。结果 :LacZ染色结果显示 ,骨骼发育早期间充质细胞聚集时Cre重组酶已经在表达Ⅱ型胶原的软骨细胞中行使功能。胚胎期 13.5d小鼠的前肢、后肢、脊椎和梅克尔软骨部位LacZ染色阳性。在新生小鼠可见由软骨内成骨形成的骨骼内软骨组织LacZ染色阳性。从新生小鼠的胫骨显微切片可以看到 ,生长板各区软骨细胞、软骨膜细胞和紧邻生长板干骺端的成骨细胞LacZ染色阳性。结论 :我们研制的Col2a1 Cre转基因小鼠中Cre重组酶在软骨内成骨过程中所有的软骨细胞中表达 ,是一种理想的研制软骨组织特异性剔除基因小鼠的工具。Objective:To further investigate the temporal and spatial distribution of the Cre recombinase in Col2a1-Cre transgenic mouse. Methods: The chondrocyte-specific Cre transgenic mice were bred with ROSA26 reporter mice. The mice carrying both Cre and ROSA26 transgenes were obtained. The activity of Cre recombinase was observed by LacZ staining in the double transgenic mice. Results: LacZ staining showed that Cre recombinase activity was observed during the period of mesenchymal cell condensation. At E13.5, the expression of Cre recombinase was correlated with the expression of collagen Ⅱ in the forelimb, hindlimb, vertebra, and Meckel′s cartilage. In the neonates, LacZ expression was detected in all cartilaginous tissues which formed through the endochondral ossification. In addition, the tibia sections of neonates showed that growth plate chondrocytes, perichondrium cells and osteoblasts adjacent to growth plate were positive for lacZ staining. Conclusion: All these data indicated that the chondrocyte-specific Cre transgenic mice specifically expressed the Cre recombinase in the cartilage tissues, and could serve as useful tool for generating chondrocyte-specific gene-knockout mice.

关 键 词：软骨细胞 CRE重组酶 小鼠 转基因 LacZ染色 

分 类 号：R346[医药卫生—基础医学]