CONSTRUCTION AND APPLICATION OF CDNA MICROARRAY SPECIALIZED FOR PERIODONTAL IIGAMENT

1. 人类牙周韧带基因表达谱分析我们收集了人类牙周韧带（PDL）的3 -directed cDNA文库序列，该文库忠实地代表了mRNA群体的组成。利用这些信息，我们研究了活性基因的表达谱。我们发现PDL具有独特的活性基因表达谱。通过分析人类PDL的a基因表达谱，我们发现了一个新基因，该基因编码一个小的富cine重复蛋白多糖家族的新成员。我们将其命名为PLAP-1（牙周韧带相关蛋白-1）。结果表明，PLAP-1在体内特异性表达于PDL中，在维持PDL - 3的稳态中起重要作用。构建人源性PDL专用cDNA芯片选择在人源性PDL中表达的基因，构建了人源性PDL专用cDNA芯片，并将其命名为PerioGen Chip，该芯片包含了从人源性PDL.4的3 -directed cDNA文库中获得的1600个cDNA克隆。利用PerioGen芯片分析PDL细胞的基因表达利用PerioGen芯片分析PDL细胞在体外分化过程中的基因表达模式。PerioGen Chip分析鉴定出15个基因在细胞分化过程中表现出双重或更多的差异表达。其中，DAN基因和Periostin基因表现出较强的转录诱导作用。我们发现DAN和Periostin在调节PDL细胞分化中起着重要作用。利用PerioGen芯片进行体内牙周组织基因表达分析我们对来自牙周病患者的牙周组织进行基因表达分析。我们能够鉴定出几个主要在牙周病变中表达的基因。这表明，PerioGen芯片可用于临床应用。少

1. Analysis of gene expression profile of human periodontal ligamentWe collected sequences with the 3 -directed cDNA library of human periodontal ligament (PDL), which faithfully represents the composition of the mRNA population. Utilizing the information, we investigated the expression profile of the active genes. We revealed that PDL has a unique active gene expression profile.2. Identification and characterization of a novel gene, PLAP-1By analyzing the a gene expression profile of human PDL, we identified a novel gene, which encodes a new member of the small let cine-rich repeat proteoglycan family. We designated it PLAP-1(periodontal ligament associated protein-1). It was shown that PLAP-1 is expressed specifically in PDL in vivo and plays an important role in maintaining homeostasis of PDL.3. Construction of customized cDNA microarray specialized for human PDLSelecting genes that are expressed in human PDL, we constructed in-house cDNA microarrays specialized for human PDL, which … More we named PerioGen Chip PerioGen Chip contained 1,600 cDNA clones obtained from the 3 -directed cDNA library of human PDL.4. Gene expression analysis of PDL utilizing PerioGen Chip Utilizing PerioGen Chipwe analyzed the patterns of gene expression during the cytodifferentiation of PDL cells in vitro. PerioGen Chip analysis identified 15 genes showing twofold or more differential expression during the cytodifferentiation. Among these genes, we focused on DAN gene and Periostin gene which showed strong induction of their transcript. We demonstrated that DAN and Periostin have crucial roles to regulate cytodifferentiation of PDL cells.5. Gene expression analysis of in vivo periodontal tissues by PerioGen ChipWe performed gene expression analysis of periodontal tissues which were derived from patients with periodontal diseases. We were able to identify several genes which were expressed predominantly in periodontal lesions. This demonstrated that PerioGen Chip can be useful for clinical application. Less