ANALYSIS OF TYPE-II COLLAGEN RNA LOCALIZATION IN CHICK WING BUDS BY INSITU HYBRIDIZATION
ANALYSIS OF TYPE-II COLLAGEN RNA LOCALIZATION IN CHICK WING BUDS BY INSITU HYBRIDIZATION
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DOI：
10.1016/0012-1606(88)90057-7
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发表时间：
1988-01-01
影响因子：
2.7
通讯作者：
SOLURSH, M
中科院分区：
文献类型：
--
作者：
SWALLA, BJ;UPHOLT, WB;SOLURSH, M
II型胶原是软骨细胞外基质的主要成分。间质分化为软骨涉及到I型胶原合成的停止和II型胶原合成的开始。用cDNA探针将鸡肢芽分离的mRNA与II型胶原mRNA进行溶液杂交，发现间充质鸡肢中存在少量的II型胶原。我们利用单链RNA探针原位杂交技术检测了II型胶原mRNa在间充质雏鸡翅芽中的定位。我们的研究结果显示，少量但可检测的II型胶原RNA均匀分布在早期四肢，直到第22期形成第一个不稳定凝聚。这很有趣，因为我们知道，从小鸡翅芽中分离出来的间充质在培养中具有成软骨的能力，即使是从肢体的非成软骨区域提取出来的。在第23期，与其他肢体细胞相比，在不稳定凝结细胞中发现II型胶原mRNA的水平显著增加。随着软骨形成的进行，II型胶原RNA的数量在软骨细胞中增加得更多。外周组织的信号与背景难以区分。这些结果表明，II型胶原蛋白信息在整个间充质鸡翼芽细胞中存在低水平，直到凝结的形成导致在肢体核心发现的软骨前细胞中II型mRNA的升高。
Type II collagen is a major component of cartilage extracellular matrix. Differentiation of mesenchyme into cartilage involves the cessation of type I collagen synthesis and the onset of type II collagen synthesis. Solution hybridization of mRNA isolated from chick limb buds with a cDNA probe to type II collagen mRNA showed the presence of small amounts of type II collagen in mesenchymal chick limbs. We have examined the localization of type II collagen mRNa in mesenchymal chick wing buds by in situ hybridization using single stranded RNA probes. Our results show a small but detectable amount of type II collagen RNA distributed uniformly in early limbs until the first precartilage condensations form at stage 22. This is interesting because it is known that mesenchyme isolated from chick wing buds has the capacity to undergo chondrogenesis in culture, even if taken from nonchondrogenic areas of the limb. At stage 23, type II collagen mRNA is found at significantly increased levels in the cells of the precartilage condensation when compared to the other limb cells. As chondrogenesis proceeds, the amount of type II collagen RNA increases even more in cells of the cartilage elements. The signal in the peripheral tissue is indistinguishable from background. These results show that type II collagen message exists at low levels in cells throughout the mesenchymal chick wing bud, until the formation of the condensation results in an elevation of type II mRNA in the prechondrogenic cells found in the core of the limb.