Differential localization of mRNAs of collagen types I and II in chick fibroblasts, chondrocytes, and corneal cells by in situ hybridization using cDNA probes
Differential localization of mRNAs of collagen types I and II in chick fibroblasts, chondrocytes, and corneal cells by in situ hybridization using cDNA probes
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使用 cDNA 探针通过原位杂交对鸡成纤维细胞、软骨细胞和角膜细胞中 I 型和 II 型胶原蛋白的 mRNA 进行差异定位
作者：
M. Hayashi;Y. Ninomiya;J. Parsons;K. Hayashi;B. Olsen;R. Trelstad
我们采用了一种高度特异性的原位杂交协议，允许差异检测的mRNA的胶原蛋白I型和II型的石蜡切片从鸡胚组织。所有探针都是编码前α链C-前肽区部分的cDNA限制性片段，并且一些片段还编码I型或II型胶原蛋白mRNA的3 '-非翻译区。肌腱成纤维细胞和胸骨软骨细胞的涂片从17天的鸡胚，以及石蜡切片的10天的整个胚胎和角膜的6.5天的胚胎与3 H-标记的探针杂交的I型或II型胶原mRNA。放射自显影显示，标记是突出的肌腱成纤维细胞与I型胶原探针和胸骨软骨细胞与II型胶原探针;在来自10-d-龄胚胎的巩膜和四肢的软骨中，I型探针显示软骨周围的成纤维细胞片和软骨中的一些软骨细胞的强标记，而II型探针强烈标记软骨细胞，仅标记少量成纤维细胞;在6.5d胚胎角膜中，I型探针标记基质中的上皮细胞和成纤维细胞较强，内皮细胞较弱，而II型探针几乎只标记上皮细胞，除了在内皮细胞中的轻微标记。这些数据表明，胚胎组织分别和/或同时表达这两种胶原基因，并提供了新的方法来研究细胞外基质成分的细胞调节。
We have employed a highly specific in situ hybridization protocol that allows differential detection of mRNAs of collagen types I and II in paraffin sections from chick embryo tissues. All probes were cDNA restriction fragments encoding portions of the C-propeptide region of the pro alpha-chain, and some of the fragments also encoded the 3'- untranslated region of mRNAs of either type I or type II collagen. Smears of tendon fibroblasts and those of sternal chondrocytes from 17- d-old chick embryos as well as paraffin sections of 10-d-old whole embryos and of the cornea of 6.5-d-old embryos were hybridized with 3H- labeled probes for either type I or type II collagen mRNA. Autoradiographs revealed that the labeling was prominent in tendon fibroblasts with the type I collagen probe and in sternal chondrocytes with the type II collagen probe; that in the cartilage of sclera and limbs from 10-d-old embryos, the type I probe showed strong labeling of fibroblast sheets surrounding the cartilage and of a few chondrocytes in the cartilage, whereas the type II probe labeled chondrocytes intensely and only a few fibroblasts; and that in the cornea of 6.5-d- old embryos, the type I probe labeled the epithelial cells and fibroblasts in the stroma heavily, and the endothelial cells slightly, whereas the type II probe labeled almost exclusively the epithelial cells except for a slight labeling in the endothelial cells. These data indicate that embryonic tissues express these two collagen genes separately and/or simultaneously and offer new approaches to the study of the cellular regulation of extracellular matrix components.
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原位杂交组织化学：一种研究单细胞基因表达的技术。
DOI：
10.1089/dna.1983.2.157
发表时间：
1983
期刊：
DNA (Mary Ann Liebert, Inc.)
影响因子：
--
作者：
Gee,CE;Roberts,JL
通讯作者：
Roberts,JL
编码软骨特异性短胶原蛋白的 cDNA 的合成和表征。
DOI：
10.1073/pnas.81.10.3014
发表时间：
1984
影响因子：
11.1
作者：
Ninomiya,Y;Olsen,BR
通讯作者：
Olsen,BR
使用原位杂交鉴定分离的小鼠肝细胞中的胶原蛋白和白蛋白 mRNA。
DOI：
10.1073/pnas.80.13.4017
发表时间：
1983
影响因子：
11.1
作者：
Saber,MA;Zern,MA;Shafritz,DA
通讯作者：
Shafritz,DA
影响因子：
--
作者：
E. J. Miller;S. Gay
通讯作者：
E. J. Miller;S. Gay
噬菌体 SP6 特异性 RNA 聚合酶。
DOI：
--
发表时间：
1982
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Butler,ET;Chamberlin,MJ
通讯作者：
Chamberlin,MJ