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人牙髓细胞中蛋白聚糖相关mRNA的鉴定

The identification of proteoglycan-associated mRNAs in human dental pulp cells.

作者信息

Harlamb S C, Messer H H

机构信息

School of Dental Science, University of Melbourne, Australia.

出版信息

Arch Oral Biol. 1996 Nov;41(11):1097-100. doi: 10.1016/s0003-9969(96)00036-2.

Abstract

Characterization of the dental pulp proteoglycans has been largely confined to the glycosaminoglycan component of the proteoglycan molecule, while the protein core has received little attention. This study was conducted to identify mRNAs of previously well-characterized proteoglycans-biglycan, decorin and versican-and link protein in dental pulp cells. Dermal fibroblasts were used as a positive control. Oligonucleotide probes were constructed based on published sequences for the four proteins from human tissues. Total RNA was isolated from cultured human pulp and dermal cells, separated according to size by formaldehyde gel electrophoresis and subsequently transferred to a nylon filter. Northern hybridizations using the oligonucleotide probes revealed the expression of biglycan, decorin, versican and link protein mRNAs. Biglycan and decorin are small proteoglycans that have a regulatory effect on collagen fibrillogenesis. Assuming expression of link protein and versican in vivo, the larger proteoglycans in the dental pulp are capable of forming large proteoglycan aggregates.

摘要

牙髓蛋白聚糖的特性研究主要局限于蛋白聚糖分子的糖胺聚糖成分,而其蛋白核心很少受到关注。本研究旨在鉴定牙髓细胞中此前已充分表征的蛋白聚糖(双糖链蛋白聚糖、核心蛋白聚糖和多功能蛋白聚糖)以及连接蛋白的信使核糖核酸(mRNA)。将真皮成纤维细胞用作阳性对照。根据已发表的人类组织中这四种蛋白质的序列构建寡核苷酸探针。从培养的人牙髓细胞和真皮细胞中分离出总RNA,通过甲醛凝胶电泳按大小分离,随后转移到尼龙滤膜上。使用寡核苷酸探针进行的Northern杂交揭示了双糖链蛋白聚糖、核心蛋白聚糖、多功能蛋白聚糖和连接蛋白mRNA的表达。双糖链蛋白聚糖和核心蛋白聚糖是对胶原纤维形成具有调节作用的小蛋白聚糖。假设连接蛋白和多功能蛋白聚糖在体内表达,牙髓中的较大蛋白聚糖能够形成大的蛋白聚糖聚集体。

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