Hofmann C, Gropp R, von der Mark K
Max Planck Society, Clinical Research Unit for Rheumatology, University of Erlangen-Nürnberg, Germany.
Dev Biol. 1992 Jun;151(2):391-400. doi: 10.1016/0012-1606(92)90179-k.
Expression of anchorin CII, a collagen-binding protein of the annexin family, was followed in the developing chick embryo using Northern and in situ hybridization and Western blotting. During chick somite development, anchorin CII mRNA was detected by Northern blotting as early as stage 11. At stage 24, anchorin mRNA accumulated in the anterior part of the somite sclerotome near the resegmentation line, as shown by in situ hybridization. The presence of anchorin CII protein during stages 11 to 20 was confirmed by Western blotting. In situ hybridization identified anchorin CII also in the otic vesicle adjacent to the site of contact with the statoacoustic ganglion and in the mandibular mesenchyme. The level of anchorin CII mRNA in differentiated hyaline cartilage, exemplified by sternal cartilage, was lower than that in differentiating somites or cultured chondrocytes. These findings are consistent with our notion that anchorin CII may be involved in cell-matrix interactions preceding chondrogenic differentiation events in the chick embryo. A significant level of anchorin CII mRNA and protein synthesis was also found in cultured myoblasts, but less than that in chondroblasts. This distribution pattern is different from that reported for a related protein, p34, or calpactin, the major protein substrate for tyrosine kinase phosphorylation in chick chondrocytes and fibroblasts. The results confirm suggestions from previous sequencing studies that anchorin CII and p34 are different proteins of the annexin/calpactin family.
使用Northern印迹法、原位杂交和蛋白质免疫印迹法,对鸡胚胎发育过程中膜联蛋白家族的一种胶原结合蛋白——锚定蛋白CII的表达进行了跟踪研究。在鸡体节发育过程中,通过Northern印迹法早在第11阶段就检测到了锚定蛋白CII的mRNA。原位杂交显示,在第24阶段,锚定蛋白mRNA在靠近重分割线的体节生骨节前部积累。通过蛋白质免疫印迹法证实了在第11至20阶段存在锚定蛋白CII蛋白。原位杂交还在与 statoacoustic 神经节接触部位相邻的耳囊中以及下颌间充质中鉴定出了锚定蛋白CII。以胸骨软骨为例,分化的透明软骨中锚定蛋白CII mRNA的水平低于分化的体节或培养的软骨细胞中的水平。这些发现与我们的观点一致,即锚定蛋白CII可能参与了鸡胚胎软骨形成分化事件之前的细胞-基质相互作用。在培养的成肌细胞中也发现存在显著水平的锚定蛋白CII mRNA和蛋白质合成,但低于成软骨细胞中的水平。这种分布模式与报道的一种相关蛋白p34或钙结合蛋白不同,后者是鸡软骨细胞和成纤维细胞中酪氨酸激酶磷酸化的主要蛋白底物。结果证实了先前测序研究的推测,即锚定蛋白CII和p34是膜联蛋白/钙结合蛋白家族中的不同蛋白。