Lin Juntang, Luo Jiankai, Redies Christoph
Institute of Anatomy I, Friedrich Schiller University Jena, Teichgraben 7, D-07743 Jena, Germany.
Brain Res. 2008 Mar 27;1201:1-14. doi: 10.1016/j.brainres.2008.01.071. Epub 2008 Feb 9.
Three types of full-length cDNAs encoding chicken cadherin-8 (Cdh8) were identified and their expression in the embryonic chicken brain was investigated. The longest type of cDNA is closely similar to that of other classic cadherins, and the predicted protein shows a high similarity to rat and human Cdh8. The second type of cDNA is considerably shorter. The deduced protein lacks the cytoplasmic tail and the transmembrane domain, and contains a truncated fifth cadherin repeat in the extracellular domain (EC5; 68 amino acids), suggesting that it is a soluble isoform. The third type resembles the second one but is even shorter (only 30 amino acids in EC5). All types contain unique short sequences at their C terminus. Genomic analysis demonstrated that all of the three Cdh8 isoforms are located on chromosome 11, and consist of 12, 10, and 9 exons, respectively. Semi-quantitative RT-PCR with type-specific primers showed that the transcription of the three Cdh8 isoforms was temporally and spatially regulated in different parts of the embryonic chicken brain. The distinct regulation of gene expression suggested that the three isoforms likely play different roles during brain development. Northern blot analysis revealed that the transcription of the long isoform was much stronger than that of the two shorter ones. In situ hybridization showed that the long isoform of Cdh8 is expressed by a specific subset of brain nuclei, regions and layers in all major parts of the brain.
我们鉴定出了三种编码鸡钙黏蛋白-8(Cdh8)的全长cDNA,并研究了它们在鸡胚胎大脑中的表达情况。最长的一种cDNA与其他经典钙黏蛋白的cDNA非常相似,预测的蛋白质与大鼠和人类的Cdh8具有高度相似性。第二种cDNA则短得多。推导的蛋白质缺乏细胞质尾巴和跨膜结构域,并且在细胞外结构域中包含一个截短的第五个钙黏蛋白重复序列(EC5;68个氨基酸),这表明它是一种可溶性异构体。第三种类型与第二种相似,但更短(EC5中只有30个氨基酸)。所有类型在其C末端都含有独特的短序列。基因组分析表明,所有三种Cdh8异构体都位于11号染色体上,分别由12、10和9个外显子组成。使用类型特异性引物进行的半定量RT-PCR表明,三种Cdh8异构体的转录在鸡胚胎大脑的不同部位受到时间和空间上的调控。基因表达的明显调控表明,这三种异构体在大脑发育过程中可能发挥不同的作用。Northern印迹分析显示,长异构体的转录比另外两种短异构体要强得多。原位杂交显示,Cdh8的长异构体由大脑所有主要部分的特定脑核、区域和层的子集表达。