Chiang R K, Barnstable C J
Department of Ophthalmology and Visual Science, Yale University School of Medicine, New Haven, CT 06520-8061, USA.
Vis Neurosci. 1998 Sep-Oct;15(5):823-9. doi: 10.1017/s0952523898155074.
The appearance of cGMP-gated cation channel protein in the postnatal rat retina has been studied by fluorescence immunocytochemistry of radial retinal sections and immunoblots of retinal membrane proteins. Channel immunoreactivity was first detectable with RCNGC1-7H2 monoclonal antibody at postnatal day 7 (PN7) by both methods. Immunocytochemical label in retinal sections was localized to the outer segments, and immunoreactivity increased with increasing age. We also compared the developmental appearance of the cGMP-gated cation channel to that of other phototransduction proteins and developmental markers. RET-P2, a monoclonal antibody recognizing the 39-kDa rds/peripherin disc protein, first labeled outer segments at PN7, coincident with cGMP-gated cation channel expression. Double labeling of the same section of PN7 rat retina with RET-P2 and R309 (a polyclonal antiserum against the rod cGMP-gated cation channel) revealed identical patterns of labelling. Similarly, double labeling with RCNGC1-7H2 and an antibody against the rod cGMP-phosphodiesterase gave coincident labeling, suggesting coordinate expression mechanisms of phototransduction proteins with each other and with outer segment structural proteins.
通过对视网膜径向切片的荧光免疫细胞化学和视网膜膜蛋白免疫印迹法，研究了产后大鼠视网膜中cGMP门控阳离子通道蛋白的出现情况。两种方法均在出生后第7天（PN7）首次用RCNGC1 - 7H2单克隆抗体检测到通道免疫反应性。视网膜切片中的免疫细胞化学标记定位于外段，且免疫反应性随年龄增长而增加。我们还比较了cGMP门控阳离子通道与其他光转导蛋白和发育标志物的发育情况。识别39 kDa rds/外周蛋白盘蛋白的单克隆抗体RET - P2在PN7首次标记外段，与cGMP门控阳离子通道表达同时出现。用RET - P2和R309（针对视杆cGMP门控阳离子通道的多克隆抗血清）对PN7大鼠视网膜同一切片进行双重标记，显示出相同的标记模式。同样，用RCNGC1 - 7H2和抗视杆cGMP磷酸二酯酶抗体进行双重标记也得到了一致的标记结果，这表明光转导蛋白之间以及与外段结构蛋白存在协同表达机制。
