Usui T, Suzuki K, Kaji Y, Amano S, Miyata K, Heldin P, Yamashita H
Department of Ophthalmology, University of Tokyo, Japan.
Invest Ophthalmol Vis Sci. 1999 Mar;40(3):563-7.
Hyaluronan (HA), a high-molecular-weight linear glycosaminoglycan, is a component of the extracellular matrix (ECM). It is expressed in eyes and plays important roles in many biologic processes, including cell migration, proliferation, and differentiation. Hyaluronan is produced by HA synthase (HAS), which has three isoforms: HAS1, HAS2, and HAS3. In this study, the HAS expression in the anterior segment of bovine eyes was investigated to determine the significance of HA in eyes.
To obtain bovine HAS probes, degenerate oligonucleotide primers, based on well-conserved amino acid sequences including the catalytic region of each HAS isoform, were used for reverse transcription-polymerase chain reaction to amplify mRNA from bovine corneal endothelial cells (BCECs). Hyaluronan synthase-1 expression in the anterior segment of bovine eyes at the protein level was investigated by immunohistochemistry.
All three HAS isoforms were expressed in BCECs at the mRNA level. Amplified cDNA fragments of HAS1, HAS2, and HAS3 from BCECs can be aligned to human counterparts, showing similarities of 100%, 97.3%, and 100%, respectively, at the amino acid level. Hyaluronan synthase 1 was expressed at the protein level in corneal epithelium, keratocyte, corneal endothelium, conjunctival epithelium, ciliary epithelium, capillary endothelium, and trabecular meshwork.
Hyaluronan synthase isoforms were expressed in the ocular anterior segment and are speculated to be involved in HA production in situ.
透明质酸(HA)是一种高分子量线性糖胺聚糖,是细胞外基质(ECM)的组成成分。它在眼部表达,并在包括细胞迁移、增殖和分化在内的许多生物学过程中发挥重要作用。透明质酸由透明质酸合酶(HAS)产生,该酶有三种同工型:HAS1、HAS2和HAS3。在本研究中,对牛眼前段的HAS表达进行了研究,以确定HA在眼中的意义。
为获得牛HAS探针,基于包括每种HAS同工型催化区域在内的保守氨基酸序列设计简并寡核苷酸引物,用于逆转录-聚合酶链反应,以扩增牛角膜内皮细胞(BCEC)中的mRNA。通过免疫组织化学研究牛眼前段透明质酸合酶-1在蛋白水平的表达。
所有三种HAS同工型在BCEC中均有mRNA水平的表达。从BCEC扩增得到的HAS1、HAS2和HAS3的cDNA片段可与人对应片段比对,在氨基酸水平上分别显示出100%、97.3%和100%的相似性。透明质酸合酶1在角膜上皮、角膜细胞、角膜内皮、结膜上皮、睫状体上皮、毛细血管内皮和小梁网中均有蛋白水平的表达。
透明质酸合酶同工型在眼前段表达,推测参与原位HA的产生。