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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 determine the temporal and spatial distribution of Cre recombinase expression in the cartilage-specific Cre recombinase transgenic mice (Col2al Cre). Methods: The gene encoding Cre recombinase will be expressed in the double transgenic mice by crossing the Col2a1 Cre transgenic mice with the ROSA2 6 reporter mice. LacZ staining can be directly observed at different developmental stages of transgenic mice Cre recombinase expression of tissue specificity. RESULTS: LacZ staining showed that Cre recombinase had been functioning in chondrocytes expressing type II collagen when early bone mesenchymal cells accumulated. Embryonic 13.5d mouse forelimb, hindlimb, spine and Merkel cartilage LacZ positive. LacZ staining was positive in skeletal cartilage tissue formed by endochondral ossification in neonatal mice. From the tibial micrographs of neonatal mice, we can see that the LacZ staining of osteoblasts of chondrocytes, perichondrocytes and metaphyseal cells in the growth plate are positive. CONCLUSION: The expression of Cre recombinase in all chondrocytes in the process of intraochondrogenic osteogenesis in our Col2a1 Cre transgenic mice is an ideal tool for developing mouse cartilage tissue-specific knockout mice.