Castagnola P, Dozin B, Moro G, Cancedda R
Laboratorio di Differenziamento Cellulare, Istituto Nazionale per la Ricerca sul Cancro, Genoa, Italy.
J Cell Biol. 1988 Feb;106(2):461-7. doi: 10.1083/jcb.106.2.461.
This report deals with the quantitation of both mRNA and transcription activity of type I collagen gene and of three cartilage-specific collagens (types II, IX, and X) during in vitro differentiation of chick chondrocytes. Differentiation was obtained by transferal to suspension culture of dedifferentiated cells passaged for 3 wk as adherent cells. The type I collagen mRNA, highly represented in the dedifferentiated cells, rapidly decreased during chondrocyte differentiation. On the contrary, types II and IX collagen mRNAs sharply increased within the first week of suspension culture, peaked in the second week, and thereafter began to decrease. This decrease was particularly significant for type IX collagen mRNA. The level of type X collagen mRNA progressively increased during the course of the culture, reached its maximal value after 3-4 wk, and decreased only at a later stage of cell differentiation. As determined by in vitro run-off transcription assays, all these changes in collagen mRNA levels could be attributed to parallel modifications in the relative rate of transcription of the corresponding collagen genes. We suggest that chicken chondrocyte differentiation proceeds through at least two different steps: (a) first, transition from a stage characterized by a high level of type I collagen mRNA to a stage characterized by predominance of types II and IX collagen mRNAs; (b) later, transition to a stage characterized by the highest level of type X collagen mRNA.
本报告探讨了鸡软骨细胞体外分化过程中I型胶原蛋白基因以及三种软骨特异性胶原蛋白(II型、IX型和X型)的mRNA定量和转录活性。分化是通过将传代3周的去分化贴壁细胞转移至悬浮培养来实现的。I型胶原蛋白mRNA在去分化细胞中高度表达,在软骨细胞分化过程中迅速减少。相反,II型和IX型胶原蛋白mRNA在悬浮培养的第一周内急剧增加,在第二周达到峰值,此后开始下降。这种下降对IX型胶原蛋白mRNA尤为显著。X型胶原蛋白mRNA水平在培养过程中逐渐升高,在3 - 4周后达到最大值,仅在细胞分化后期下降。通过体外径流转录分析确定,胶原蛋白mRNA水平的所有这些变化都可归因于相应胶原蛋白基因转录相对速率的平行改变。我们认为鸡软骨细胞分化至少经历两个不同阶段:(a)首先,从以高水平I型胶原蛋白mRNA为特征的阶段转变为以II型和IX型胶原蛋白mRNA占主导为特征的阶段;(b)随后,转变为以X型胶原蛋白mRNA水平最高为特征的阶段。