Balmain N, Leguellec D, Elkak A, Nars G, Toury R, Schoevaert D
INSERM Unité 120, Hôpital Robert Debré, Paris, France.
Cell Mol Biol (Noisy-le-grand). 1995 Feb;41(1):197-212.
The spatial-temporal distribution of the mRNAs for type IX and type XI collagens were compared to that of type II collagen mRNA in the tibial epiphyseal plate cartilage of normal growing rats. The mRNAs were detected by in situ hybridization with radio-labelled specific probes and visualized by radioautography. The areas covered by the resulting silver grains were quantified by computer assisted image analysis. The areas in chondrocytes of each zone of the epiphyseal plate cartilage, which correspond to the stages of chondrocyte development and function were determined. Types II, IX and XI mRNAs were present to some extent in chondrocytes of all zones. The distributions of type II and type IX collagen mRNAs were similar with the highest concentrations in the proliferative zone, and the lowest in the resting and calcifying zones chondrocytes. In contrast, type XI collagen mRNA had a different distribution, with the lowest concentration in the resting zone chondrocytes and a significant decrease in the calcifying zone chondrocytes. These patterns correlates with the changes in chondrocyte function, and may reflect the roles of the type IX and type XI collagens. The data show that computer assisted image analysis of in situ hybridization radioautographic images is a precise, rapid tool for analysing differences in gene expression.
在正常生长大鼠的胫骨骨骺板软骨中，比较了IX型和XI型胶原蛋白mRNA与II型胶原蛋白mRNA的时空分布。通过用放射性标记的特异性探针进行原位杂交检测mRNA，并通过放射自显影进行可视化。通过计算机辅助图像分析对所得银颗粒覆盖的面积进行定量。确定了骨骺板软骨各区域软骨细胞中的面积，这些区域对应于软骨细胞发育和功能的阶段。II型、IX型和XI型mRNA在所有区域的软骨细胞中均有一定程度的存在。II型和IX型胶原蛋白mRNA的分布相似，在增殖区浓度最高，在静止区和钙化区软骨细胞中浓度最低。相比之下，XI型胶原蛋白mRNA具有不同的分布，在静止区软骨细胞中浓度最低，在钙化区软骨细胞中显著降低。这些模式与软骨细胞功能的变化相关，可能反映了IX型和XI型胶原蛋白的作用。数据表明，对原位杂交放射自显影图像进行计算机辅助图像分析是一种精确、快速的分析基因表达差异的工具。
