Tanabe Kazuhiko, Kimura Itaru, Okamoto Haru, Chi Zai-Long, Akahori Masakazu, Shimozawa Nobuhiro, Ebihara Nobuyuki, Murakami Akira, Iwata Takeshi
Juntendo Iji Zasshi. 2022 Jul 14;68(4):339-351. doi: 10.14789/jmj.JMJ21-0042-OA. eCollection 2022.
The purpose of this study was to determine what proteins are present in the ciliary body (CB). To accomplish this, we conducted a proteomic analysis of the CB of cynomolgus monkeys. We also determined the location of the proteins in CB by immunohistology.
The eyes of euthanized cynomolgus monkeys were enucleated, and the CB, were isolated from the eyes. Proteins were extracted from the CB and determined by liquid chromatography-mass spectrometry. Separated CB epithelial cells were cultured, and the proteins expressed in the CB were determined by Western blotting. The location of these proteins in the CB was determined by immunohistochemical staining. We also investigated whether adding dexamethasone to the culture medium changed protein expression by the epithelial cells.
Proteomic analysis of the CBs showed that 813 proteins were expressed in the epithelium and stroma. These proteins included the small guanosine triphosphate-binding protein Rab8 and the ezrin/radixin/moesin (ERM) family. Tissue and immunohistological staining confirmed the colocalization of these proteins in non-pigmented CB epithelium. Western blotting of cultured CB epithelial cell lysates showed a tendency that adding dexamethasone changed Rab8 protein expression levels.
Proteomic analysis of CBs identified several proteins involved in the transport and secretion of proteins. These proteins may be involved in the production of aqueous humor and protein secretion by the CB.
本研究的目的是确定睫状体（CB）中存在哪些蛋白质。为实现这一目标，我们对食蟹猴的睫状体进行了蛋白质组学分析。我们还通过免疫组织学确定了这些蛋白质在睫状体中的位置。
摘除安乐死的食蟹猴的眼睛，从眼中分离出睫状体。从睫状体中提取蛋白质，并通过液相色谱 - 质谱法进行测定。培养分离出的睫状体上皮细胞，通过蛋白质印迹法测定睫状体中表达的蛋白质。通过免疫组织化学染色确定这些蛋白质在睫状体中的位置。我们还研究了向培养基中添加地塞米松是否会改变上皮细胞的蛋白质表达。
睫状体的蛋白质组学分析表明，上皮和基质中表达了813种蛋白质。这些蛋白质包括小GTP结合蛋白Rab8和埃兹蛋白/根蛋白/膜突蛋白（ERM）家族。组织和免疫组织学染色证实了这些蛋白质在非色素睫状体上皮中的共定位。培养的睫状体上皮细胞裂解物的蛋白质印迹显示，添加地塞米松有改变Rab8蛋白表达水平的趋势。
睫状体的蛋白质组学分析鉴定了几种参与蛋白质转运和分泌的蛋白质。这些蛋白质可能参与房水的产生和睫状体的蛋白质分泌。
