Yamada S, Murakami S, Matoba R, Ozawa Y, Yokokoji T, Nakahira Y, Ikezawa K, Takayama S, Matsubara K, Okada H
Department of Periodontology, Osaka University Graduate School of Dentistry, 1-8 Yamadaoka, Suita, Osaka 565-0871, Japan.
Gene. 2001 Sep 19;275(2):279-86. doi: 10.1016/s0378-1119(01)00683-7.
Periodontal ligament (PDL) is one of the most important tissues in maintaining the homeostasis of tooth and tooth-supporting tissue, periodontium. In this study, we investigated the expression profile of active genes in the human PDL obtained by collecting sequences with a 3'-directed cDNA library, which faithfully represents the composition of the mRNA population. We succeeded in obtaining a total of 1752 cDNA sequences by sequencing randomly selected clones and identified a total of 1318 different species as gene signatures (GS) by their sequence identity, 344 of which were known genes in the GenBank, and 974 of which were new genes. The resulting expression profile showed that collagen type I and type III were the most abundant genes and that osteogenesis-related proteins, such as SPARC/osteonectin and osteoblast specific factor 2, were highly expressed. By comparing the expression profile of PDL with 44 profiles similarly obtained with unrelated human cell/tissue, nine novel genes, which are probably expressed specifically in PDL, were discovered. Among them, we cloned a full-length cDNA of GS5096, which is frequently expressed in freshly-isolated periodontal tissue. We found that it encodes a novel protein, which is a new member of the class I small leucine-rich repeat proteoglycan family, and designated it PLAP-1 (periodontal ligament associated protein-1). PLAP-1 mRNA expression was confirmed in in vitro-maintained PDL cells and was enhanced during the course of the cytodifferentiation of the PDL cells into mineralized tissue-forming cells such as osteoblasts and cementoblasts. These findings suggest the involvement of PLAP-1 in the mineralized matrix formation in PDL tissues.
牙周韧带(PDL)是维持牙齿及牙齿支持组织——牙周组织内环境稳定的最重要组织之一。在本研究中,我们通过构建3'端定向cDNA文库收集序列,研究了人牙周韧带中活性基因的表达谱,该文库能忠实地反映mRNA群体的组成。通过对随机挑选的克隆进行测序,我们成功获得了总共1752个cDNA序列,并根据序列同一性鉴定出总共1318个不同的物种作为基因特征(GS),其中344个是GenBank中的已知基因,974个是新基因。所得的表达谱显示,I型和III型胶原蛋白是最丰富的基因,并且与成骨相关的蛋白质,如富含半胱氨酸的酸性分泌蛋白/骨连接蛋白和成骨细胞特异性因子2,表达水平很高。通过将牙周韧带的表达谱与用无关的人类细胞/组织类似获得的44个表达谱进行比较,发现了9个可能在牙周韧带中特异性表达的新基因。其中,我们克隆了GS5096的全长cDNA,其在新鲜分离的牙周组织中频繁表达。我们发现它编码一种新蛋白质,它是富含亮氨酸的小分子重复蛋白聚糖I类家族的新成员,并将其命名为PLAP-1(牙周韧带相关蛋白-1)。PLAP-1 mRNA的表达在体外培养的牙周韧带细胞中得到证实,并且在牙周韧带细胞向矿化组织形成细胞如成骨细胞和牙骨质细胞的细胞分化过程中增强。这些发现表明PLAP-1参与了牙周韧带组织中矿化基质的形成。