Yuan Xiaoyong, Chen Zhuo, Yang Zhuo, Gao Junfang, Zhang Aijun, Wu Samuel M, Jacoby Roy
Department of Ophthalmology, Cullen Eye Institute, Baylor College of Medicine, Houston, TX 77030, USA.
Cornea. 2009 Feb;28(2):194-9. doi: 10.1097/ICO.0b013e318185268e.
To detect the expression pattern of connexins in epithelial cells of the central cornea and limbus of the macaque.
Total RNA was extracted from the central corneal and limbal epithelia of Macaca fascicularis and processed by reverse transcriptase-polymerase chain reaction with isoform primers to detect the expression of 16 connexin (Cx). Immunofluorescent staining of frozen sections of corneal tissue confirmed and localized connexin proteins expression.
Transcripts encoding 10 Cx isoforms (Cx26, Cx30, Cx30.3, Cx31, Cx31.1, Cx32, Cx43, Cx45, Cx50, and Cx58) were detected by reverse transcriptase-polymerase chain reaction in both central and peripheral corneal epithelium. Six (Cx26, Cx31, Cx32, Cx43, Cx45, and Cx58) were confirmed by laser scanning confocal microscopy. Cx26 was detected throughout the central corneal epithelium and in the mid and superficial layers of the limbal epithelium. Cx43 and Cx45 were localized to the basal and suprabasal epithelial cells. Cx58 was expressed in the superficial epithelium throughout the cornea, whereas Cx31 and Cx32 were mainly expressed in the central corneal epithelium and weakly in the limbal area.
The complex distribution pattern of the connexins suggests that selected isoforms play important roles in maintaining corneal homeostasis.
检测猕猴中央角膜和角膜缘上皮细胞中连接蛋白的表达模式。
从食蟹猴的中央角膜和角膜缘上皮中提取总RNA，并用同工型引物通过逆转录-聚合酶链反应进行处理，以检测16种连接蛋白（Cx）的表达。角膜组织冰冻切片的免疫荧光染色证实并定位了连接蛋白的表达。
通过逆转录-聚合酶链反应在中央和周边角膜上皮中均检测到编码10种Cx同工型（Cx26、Cx30、Cx30.3、Cx31、Cx31.1、Cx32、Cx43、Cx45、Cx50和Cx58）的转录本。激光扫描共聚焦显微镜证实了其中6种（Cx26、Cx31、Cx32、Cx43、Cx45和Cx58）。Cx26在整个中央角膜上皮以及角膜缘上皮的中层和表层均有检测到。Cx43和Cx45定位于基底和基底上层上皮细胞。Cx58在整个角膜的表层上皮中表达，而Cx31和Cx32主要在中央角膜上皮中表达，在角膜缘区域表达较弱。
连接蛋白的复杂分布模式表明，特定的同工型在维持角膜内环境稳定中起重要作用。
