Treilleux I, Mallein-Gerin F, Le Guellec D, Bouvier R, Berger F, Herbage D
Institut de Biologie et Chimie des Protéines, Université Claude Bernard Lyon I, Villeurbanne.
Ann Pathol. 1991;11(1):47-53.
Human skeletal tissues fixed in Bouin's solution and embedded in paraffin were studied in order to identify cells responsible for production of types I and II collagens by in situ hybridization. The probes used were a double stranded cDNA fragment (type I collagen) and a synthetic oligonucleotide (type II collagen). The specificity of these probes labeled with 32P was proven in hybridizations to sections of human fetal femoral heads: fibroblasts and chondrocytes, known to produce respectively type I and type II collagen were only recognized by the corresponding probes. These results suggest that paraffin blocks available in pathology departments could be reexamined by in situ hybridization. Furthermore, the use of synthetic oligonucleotides without cloning the gene of interest could increase the usefulness of the technique in studies on human cartilage diseases and possibly on other diseases.
为了通过原位杂交鉴定负责产生I型和II型胶原蛋白的细胞,研究了固定在Bouin溶液中并包埋在石蜡中的人体骨骼组织。所使用的探针是双链cDNA片段(I型胶原蛋白)和合成寡核苷酸(II型胶原蛋白)。这些用32P标记的探针的特异性在与人类胎儿股骨头切片的杂交中得到证实:已知分别产生I型和II型胶原蛋白的成纤维细胞和软骨细胞仅被相应的探针识别。这些结果表明,病理科可用的石蜡块可通过原位杂交重新检查。此外,在不克隆感兴趣基因的情况下使用合成寡核苷酸可以提高该技术在人类软骨疾病研究以及可能在其他疾病研究中的实用性。