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目的 :用原位杂交法研究骨和软骨组织中 Ⅰ、Ⅱ 型胶原基因的表达。方法 :以与 Ⅰ 型、ⅡA型和 ⅡB型胶原mRNA特异性互补的寡核苷酸序列为探针 ,用末端转移酶将地高辛标记于 3′ -末端 ,在兔胫骨近端组织切片上进行Northen原位杂交 ,检测关节软骨、骺板和新生骨小梁区三种胶原基因的表达情况。结果 :关节软骨和骺板区均检测到软骨细胞中有 ⅡB型胶原基因表达 ,新生骨小梁区检测到成骨细胞中有 Ⅰ型胶原基因表达。结论 :本实验方法灵敏度高 ,结果可靠 ,是研究与骨病和骨发生发育相关的胶原基因表达的有力工具。
Objective: To study the expression of type Ⅰ and type Ⅱ collagen in bone and cartilage by in situ hybridization. Methods: Digoxigenin was labeled on the 3 ’- end with terminal transferase by using oligonucleotide sequence specific to type Ⅰ, type ⅡA and ⅡB collagen mRNA as probe. Northen in situ hybridization was performed to detect the expression of three collagen genes in articular cartilage, epiphyseal plate and newborn trabecular bone region. Results: Type ⅡB collagen gene was detected in chondrocytes and articular cartilage and epiphyseal plate. Type Ⅰ collagen gene expression in osteoblasts was detected in the new trabecular bone region. Conclusion: The experimental method is highly sensitive and reliable, which is a powerful tool to study collagen gene expression related to bone disease and bone development.