Prominin1在C57BL/6小鼠视网膜色素上皮中的表达

Expression of Prominin1 in retinal pigment epithelium of C57BL/6 mice at different development stages

目的探究Prominin1在不同发育时期小鼠视网膜色素上皮(retinal pigment epithelium,RPE)的表达。方法孕鼠和出生后的Prom1cre/ERT2+/Gt(ROSA)26SorCAG-Zs Green1+转基因小鼠,腹腔注射他莫昔芬,将视网膜组织冰冻切片后,采用激光共聚焦显微镜检测Prominin1在转基因小鼠RPE中的表达;利用荧光定量PCR和免疫荧光技术检测Prominin1在野生型小鼠不同发育时期RPE中的表达。结果对13.5 d(E13.5)、15.5 d(E15.5)和出生后1 d(P1)小鼠的研究结果显示:Prominin1可能在胎鼠的部分RPE表达,出生后小鼠不表达。定量PCR显示:Prominin1在胎鼠RPE(E10.5、E13.5、E15.5和E17.5)表达明显高于出生后(P1和P14)(P<0.01);在胚胎时期,Prominin1在E13.5和E15.5的RPE表达明显高于其他时期(P<0.05或P<0.01)。免疫荧光染色结果显示,Prominin1在E10.5、E13.5、E15.5和E17.5的RPE表达,而在P1不表达。结论 Prominin1在胎鼠视网膜色素上皮中表达,在E13.5和E15.5的胎鼠中尤为明显。

Objective To investigate the expression of Prominin1 in Retinal Pigment Epithelium(RPE)of mice at differential development stages.Methods Tamoxifen was intraperitoneally injected in pregnant or postnatal cre/ERT2+/CAG-ZsGreen1+transgenic mice.The expression of Prominin1 in RPE of transgenic mice was examined by confocal laser scanning microscope with frozen section.In addition,the expression of Prominin 1 in developmental RPE of wild type mice was detected with real-time PCR and immunofluorescence.Results Prominin1was expressed in RPE of E13.5,E15.5 of transgenic mice tracer,however,not expressed in postnatal mice(P1).Real-time PCR showed that the expression of prominin1 was significantly higher in embryonic stages(E10.5,E13.5,E15.5 and E17.5)than that in postnatal stages(P<0.05).Moreover,the expression of Prominin1 in RPE at E13.5 and E15.5 was significantly higher than that in other embryonic periods(P<0.05 or P<0.01).Immunofluorescence showed that Prominin1 was expressed at E10.5,E13.5,E15.5 and E17.5,but not expressed at P1.Conclusion Prominin 1 is expressed in RPE at different embryonic stages of mice,especially,at E13.5 and E15.5.