Type II collagen mRNA containing an alternatively spliced exon predominates in the chick limb prior to chondrogenesis.
Type II collagen mRNA containing an alternatively spliced exon predominates in the chick limb prior to chondrogenesis.
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DOI：
10.1016/s0021-9258(18)54517-8
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发表时间：
1991-12
期刊：
影响因子：
--
通讯作者：
Hyun-Duck Nah;W. B. Upholt
中科院分区：
文献类型：
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作者：
Hyun-Duck Nah;W. B. Upholt
利用单侧聚合酶链反应技术，获得了鸡II型胶原mRNA 5'端的一系列cDNA克隆。这些cDNA的分析表明，该基因的第二个外显子是可变剪接的，因此它在mRNA中存在或不存在。该外显子编码一个70个氨基酸的富含半胱氨酸的球状结构域，该结构域存在于α 1（I）、α 1（III）和α 2（V）前胶原链的氨基末端前肽中，但以前认为II型前胶原中不存在。鸡II型胶原mRNA的两种选择性剪接形式的表达的分析表明，没有第二个外显子的mRNA是主要形式（约90%），在胸骨软骨从14天的胚胎，但在前软骨肢间充质只有包括第二个外显子的形式被检测到。这种后一种形式也存在于许多非软骨组织中，包括胚胎颅骨、皮肤、心脏、骨骼肌和大脑;在肝脏中未检测到II型胶原mRNA。从进行性胚胎阶段发育肢体的研究表明，缺乏第二个外显子的mRNA的外观是一个相对较晚的事件在软骨形成。
A series of cDNA clones corresponding to the 5' end of the chicken type II collagen mRNA were generated using a single-sided polymerase chain reaction technique. Analysis of these cDNAs showed that the second exon of the gene is alternatively spliced such that it is either present or absent in the mRNA. This exon encodes a 70-amino acid cysteine-rich globular domain which is present in the amino-terminal propeptides of alpha 1(I), alpha 1(III), and alpha 2(V) procollagen chains but which was previously thought to be absent from type II procollagen. Analysis of the expression of the two alternatively spliced forms of the chicken type II collagen mRNA showed that the mRNA without the second exon was the predominant form (approximately 90%) in sternal cartilage from 14-day embryos, but in precartilage limb mesenchyme only the form including the second exon was detected. This later form was also present in a number of non-cartilage tissues including embryonic calvaria, skin, heart, skeletal muscle, and brain; no type II collagen mRNA was detected in liver. Studies of developing limbs from progressive embryonic stages suggest that the appearance of the mRNA lacking the second exon is a relatively late event during chondrogenesis.