Cystic Fibrosis/Pulmonary Research and Treatment Center, University of North Carolina, Chapel Hill, 7011 Thurston-Bowles Building, 96 Manning Drive, NC 27599, United States.
Exp Eye Res. 2012 Oct;103:22-32. doi: 10.1016/j.exer.2012.07.005. Epub 2012 Jul 22.
The aim of the study was to elucidate aquaporin (AQP) family member mRNA expression and protein expression/localization in the rat lacrimal functional unit. The mRNA expression of all rat AQPs (AQP0-9, 11-12) in palpebral, fornical, and bulbar conjunctiva, cornea, lacrimal gland, and Meibomian gland was measured by Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) and real time RT-PCR. Antibodies against AQP1, 3, 4, 5, 9, and 11 were used in Western blotting and immunohistochemistry to determine protein expression and distribution. Our study demonstrated characteristic AQP expression profiles in rat ocular tissues. AQP1, 3, 4, 5, 8, 9, 11, and 12 mRNA were detected in conjunctiva. AQP0, 1, 2, 3, 4, 5, 6, 11, and 12 mRNA were expressed in cornea. AQP0, 1, 2, 3, 4, 5, 7, 8, and 11 mRNA were detected in lacrimal gland. AQP1, 3, 4, 5, 7, 8, 9, 11, and 12 mRNA were identified in Meibomian gland. By Western blot, AQP1, 3, 5, and 11 were detected in conjunctiva; AQP1, 3, 5, and 11 were identified in cornea; AQP1, 3, 4, 5, and 11 were detected in lacrimal gland; and AQP1, 3, 4, 5, 9, and 11 were present in Meibomian gland. Immunohistochemistry localized AQPs to distinct sites in the various tissues. This study rigorously analyzed AQPs expression and localization in rat conjunctiva, cornea, lacrimal gland, and Meibomian gland tissues. Our findings provide a comprehensive platform for further investigation into the physiological or pathophysiological relevance of AQPs in ocular surface.
本研究旨在阐明水通道蛋白(AQP)家族成员在大鼠泪液功能单位中的 mRNA 表达和蛋白表达/定位。通过逆转录-聚合酶链反应(RT-PCR)和实时 RT-PCR 测量了大鼠所有 AQP(AQP0-9、11-12)在睑结膜、穹窿结膜和球结膜、角膜、泪腺和睑板腺中的 mRNA 表达。使用针对 AQP1、3、4、5、9 和 11 的抗体进行 Western blot 和免疫组织化学,以确定蛋白质表达和分布。我们的研究表明,大鼠眼组织具有特征性的 AQP 表达谱。在结膜中检测到 AQP1、3、4、5、8、9、11 和 12 mRNA。在角膜中表达 AQP0、1、2、3、4、5、6、11 和 12 mRNA。在泪腺中检测到 AQP0、1、2、3、4、5、7、8 和 11 mRNA。在睑板腺中鉴定出 AQP1、3、4、5、7、8、9、11 和 12 mRNA。通过 Western blot,在结膜中检测到 AQP1、3、5 和 11;在角膜中鉴定出 AQP1、3、5 和 11;在泪腺中检测到 AQP1、3、4、5 和 11;在睑板腺中存在 AQP1、3、4、5、9 和 11。免疫组织化学将 AQP 定位到各种组织中的不同部位。本研究严格分析了大鼠结膜、角膜、泪腺和睑板腺组织中 AQP 的表达和定位。我们的发现为进一步研究 AQP 在眼表面的生理或病理生理相关性提供了一个全面的平台。