Abstract：
The trabecular meshwork (TM) from primary open-angle glaucoma (POAG) cases has been found to contain decreased levels of intracellular plasmalogens. Plasmalogens are a subset of lipids involved in diverse cellular processes such as intracellular signaling, membrane asymmetry, and protein regulation. Proper plasmalogen biosynthesis is regulated by rate-limiting enzyme fatty acyl-CoA reductase (Far1). ATPase phospholipid transporting 8B2 (ATP8B2) is a type IV P-type ATPase responsible for the asymmetric distribution of plasmalogens between the intracellular and extracellular leaflets of the plasma membranes. Here we describe the methodology for extraction and culturing of TM cells from corneal tissue and subsequent downregulation of ATP8B2 using siRNA transfection. Further quantification and downstream effects of ATP8B2 gene knockdown will be analyzed utilizing immunoblotting techniques.
摘要：
已发现原发性开角型青光眼（POAG）病例的小梁网（TM）含有降低的细胞内疟原虫水平。疟原虫是参与多种细胞过程的脂质的一个子集，例如细胞内信号传导。膜不对称，和蛋白质调节。适当的缩醛磷脂生物合成受限速酶脂酰辅酶A还原酶(Far1)调节。ATPase磷脂转运8B2（ATP8B2）是IV型P型ATPase，负责在质膜的细胞内和细胞外小叶之间不对称分布。在这里，我们描述了从角膜组织提取和培养TM细胞以及随后使用siRNA转染下调ATP8B2的方法。使用免疫印迹技术分析ATP8B2基因敲低的进一步定量和下游效应。
