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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 Using oligonucleotide sequence specific to type Ⅰ, type ⅡA and type Ⅱ collagen mRNA as probe, digoxigenin was labeled with 3 ’- end by terminal transferase and was performed on the proximal tibial tissue of rabbits Northen in situ hybridization, detection of articular cartilage, epiphyseal plate and neonatal trabecular bone three collagen gene expression. Results The expression of collagen type ⅡB in chondrocytes was detected in both articular cartilage and epiphyseal plate. The type Ⅰ collagen gene expression in osteoblasts was detected in the new trabecular bone region. Conclusion The experimental method has high sensitivity and reliable results and is a powerful tool for studying collagen gene expression related to bone disease and bone development