Over the past few years, considerable progress has been achieved about the extracellular elements and intracellular regulatory molecules that are involved in the regulation of chondrogenesis. However, little is known about the molecular mechanism of how these molecules influence the gene activities during cartilage differentiation. Recently we isolated a Chicken Cartilage derived Matrix Protein (CCMP 10), a novel protein, from chicken prechondrogenic mesenchyme. To further understand the function of CCMP-10 in cartilage development, we investigated the expression of CCMP-10 during the prechondrocyte differentiation in chick embryos and micromass cultured prechondrogenic cells, using a variety of methods such as transient transfection of CCMP 10, immunohistochemical localization, northern analysis, and western analysis. When transiently transfected, CCMP 10 was expressed in both nucleus and cytoplasm, with stronger intensity in the nucleus. In an immunohistochemical study, CCMP 10 was expressed in prechondrogeinc mesenchymal cell, perichondrium, and resting and proliferative zone of the growth plate of long bone, while no expression of CCMP 10 was observed in upper mature chondrocytes and hypertrophic chondrocytes. Northern analysis of micromass cultured prechondrogenic cells showed the expression of CCMP-10 mRNA for first 2 days, while Col 2a1, aggrecan, and CMP mRNAs, known genes to express in mature chondrocyte, initiated the expression at day 2 and continued to express by day 5. In western analysis, CCMP-10 was detected at initial stage and continued to express by day 3, while Col 2al protein began to express only one day after, and continued to express. Taken together, our data suggest that CCMP-10 may play a significant role in the early cartilage development.
Aggrecans ; Animals ; Cartilage* ; Chick Embryo ; Chickens* ; Chondrocytes ; Chondrogenesis* ; Cytoplasm ; Growth Plate ; Mesoderm ; RNA, Messenger ; Transfection