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作 者:邵志华[1] 李瑛泽 许洁[1] 于金军 刘志学[3] 李思光[1] SHAO Zhi-hua;LI Ying-ze;XU Jie;YU Jin-jun;LIU Zhi-xue;LI Si-guang(Tongji University School of Medicine,Shanghai 200092,China;College of Life Sciences,Nanchang University,Nanchang 330031,China;School of Life Sciences and Technology,Tongji University,Shanghai 200092,China)
机构地区:[1]同济大学医学院,上海200092 [2]南昌大学生命科学学院,南昌330031 [3]同济大学生命科学与技术学院,上海200092
出 处:《同济大学学报(医学版)》2018年第1期23-28,共6页Journal of Tongji University(Medical Science)
基 金:国家自然科学基金(31571405)
摘 要: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.
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