Lee S K, Malpeli M, Cancedda R, Utani A, Yamada Y, Kleinman H K
Craniofacial Developmental Biology and Regeneration Branch, National Institute of Dental Research, National Institutes of Health, Bethesda, Maryland 20892-4370, USA.
Exp Cell Res. 1997 Oct 10;236(1):212-22. doi: 10.1006/excr.1997.3710.
We have observed that laminins are expressed in the chondrocytes of chick embryo sternum, mouse limb bud, and adult mouse knee joint by the methods of in situ hybridization, immunohistochemistry, Western blotting, and immunoprecipitation. From in situ hybridization using similar sized RNA probes for different mouse laminin chains, mRNAs for the alpha 1, alpha 2, beta 1, beta 2, and gamma 1 chains were expressed in the chondrocytes of chick embryo sternum, mouse limb bud, and the articular cartilage cap and epiphyseal growth plate of adult mouse knee joint. Through the use of chain-specific antibodies, staining for laminins was observed in the cytoplasm of chondrocytes from chick embryo sternum, mouse limb bud, and adult mouse knee joint. Western blot analysis confirmed the presence of laminin chains in the cells and sternal tissues. Cultured chick embryonic sternal chondrocytes expressed laminin mRNAs in the proliferating stage (2-3 days of culture) but the level increased in the aggregated cells during the maturation stage (5-7 days of culture). Comparable data were also obtained after immunostaining the cells. Thus, laminins are expressed in significant amounts by chondrocytes and may have an important role in cartilage development.
我们通过原位杂交、免疫组织化学、蛋白质印迹和免疫沉淀等方法观察到,层粘连蛋白在鸡胚胸骨、小鼠肢芽和成年小鼠膝关节的软骨细胞中表达。使用针对不同小鼠层粘连蛋白链的大小相似的RNA探针进行原位杂交,结果显示,α1、α2、β1、β2和γ1链的mRNA在鸡胚胸骨、小鼠肢芽以及成年小鼠膝关节的关节软骨帽和骨骺生长板的软骨细胞中表达。通过使用链特异性抗体,在鸡胚胸骨、小鼠肢芽和成年小鼠膝关节的软骨细胞胞质中观察到层粘连蛋白的染色。蛋白质印迹分析证实细胞和胸骨组织中存在层粘连蛋白链。培养的鸡胚胸骨软骨细胞在增殖期(培养2 - 3天)表达层粘连蛋白mRNA,但在成熟阶段(培养5 - 7天)聚集细胞中的表达水平升高。对细胞进行免疫染色后也获得了类似的数据。因此,软骨细胞大量表达层粘连蛋白,其可能在软骨发育中发挥重要作用。