Comparison of EphA receptor tyrosine kinases and ephrinA ligand expression to EphB-ephrinB in vascularized corneas
Comparison of EphA receptor tyrosine kinases and ephrinA ligand expression to EphB-ephrinB in vascularized corneas
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DOI：
10.1097/ico.0b013e3180335526
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发表时间：
2007-06-01
期刊：
影响因子：
2.8
通讯作者：
Azar, Dimitri T.
中科院分区：
文献类型：
--
作者：
Kojima, Takashi;Chung, Tae-Young;Azar, Dimitri T.
目的：Eph细胞表面受体及其配体ephrin参与神经元模式和新生血管的形成。我们的目的是比较和表征ephrinA配体和EphA受体对ephrinB的表达；配体和EphB受体在切除的小鼠角膜组织，角膜上皮和角质细胞系，以及角膜血管生成过程中。方法：采用SV40T抗原病毒感染原代培养物，使小鼠角膜上皮细胞和角质细胞永生化。克隆永生化上皮细胞和角化细胞，并使用角蛋白、波形蛋白、整合素α 5 β 1和α -平滑肌肌动蛋白抗体对其进行鉴定。植入碱性成纤维细胞生长因子微球诱导角膜新生血管形成。取野生型、ephrinB2（tlacZ/+）和EphB4（tlacZ/+）杂合小鼠的眼睛，在植入微球7天后进行切片。采用共聚焦免疫组化方法比较Eph/ephrinA家族（EphA1-8, ephrinA1-5）和Eph/ephrinB家族（EphB1-4, EphB6 ephrinB1-3）的表达。结果：野生型小鼠角膜上皮细胞和角膜形成细胞中均检测到EphA1、EphA3、ephrinA1、ephrinA2、EphB1、EphB4、ephrinB1和ephrinB2。EphA2仅在上皮细胞中免疫定位。EphA3、ephrinA1、EphB1、EphB4和ephrinB1免疫定位于角膜上皮和基质。在血管化角膜中，ephrinB1主要免疫定位于血管周围的角化细胞，ephrinB2、ephb1和EphB4主要与CD31共定位于血管内皮细胞。结论：本研究对ephrin配体和Eph受体在角膜血管生成过程中的表达进行了表征，提示Eph/ephrin受体酪氨酸激酶家族及其配体可能在角膜血管生成过程中发挥调控作用。
Purpose: Eph cell surface receptors and their ligands, ephrins, are involved in neuronal patterning and neovascularization. Our purpose is to compare and characterize the expression of ephrinA ligands and EphA receptors to ephrinB; ligands and EphB receptors in excised mouse corneal tissue, in corneal epithelial and keratocyte cell lines, and during corneal angiogenesis.Methods: Mouse corneal epithelial cells and keratocytes were immortalized using SV40T antigen viral infection of primary cultures. The immortalized epithelial cells and keratocytes were cloned and characterized using antibodies to keratin, vimentin, integrin alpha 5 beta 1, and alpha-smooth muscle actin. Basic fibroblast growth factor pellets were implanted to induce corneal neovascularization. The eyes of wild-type, ephrinB2(tlacZ/+), and EphB4(tlacZ/+) heterozygous mice were harvested and sectioned 7 days after pellet implantation. Confocal immunohistochemistry was performed to compare the expression of the Eph/ephrinA family (EphA1-8, ephrinA1-5) and Eph/ephrinB family (EphB1-4, EphB6 ephrinB1-3).Results: EphA1, EphA3, ephrinA1, ephrinA2, EphB1, EphB4, ephrinB1, and ephrinB2 were detected in wild-type mouse corneal epithelial cells and keratocytes. EphA2 was immunolocalized only in epithelial cells. Also, EphA3, ephrinA1, EphB1, EphB4, and ephrinB1 were immunolocalized to the comeal epithelium and stroma. In the vascularized corneas, ephrinB1 was immunolocalized mainly to the keratocytes around the vessels, and ephrinB2, EphB 1, and EphB4 were colocalized mainly with CD31 to the vascular endothelial cells.Conclusions: The characterization of ephrin ligand and Eph receptor expression during cornea angiogensis in this study suggests that the Eph/ephrin family of receptor tyrosine kinases and their ligands may play a role in the regulation of corneal angiogenesis.