小鼠Cpne5基因mRNA水平的表达分布及其在胚胎的表达定位  

Expression of Mouse Cpne5 Gene mRNA in Various Tissues and Its Location in Embryo

作　　者：靳雁斌[1] 丁学锋[2] 司慧远[1] 吴彦瑞 王晓文[2] 吴燕[2] 范文红[2] 

机构地区：[1]空军总医院,北京100142 [2]军事医学科学院基础医学研究所认知科学研究室,北京100850 [3]解放军第一八八医院,广东潮州521000

出　　处：《标记免疫分析与临床》2013年第2期93-96,共4页Labeled Immunoassays and Clinical Medicine

基　　金：国家自然科学基金青年基金(No.31200822);北京市自然科学基金面上项目(No.5122033)联合资助

摘　　要：目的明确小鼠Cpne5基因在mRNA水平的组织表达分布及其在胚胎的表达定位。方法提取新生、成年小鼠主要脏器的总RNA,利用RT-PCR法检测Cpne5 mRNA的表达量;合成针对Cpne5 mRNA的杂交探针,取不同胎龄胎鼠进行整体原位杂交。结果 Cpne5 mRNA在成年小鼠10种脏器组织均有不同程度的表达,以大脑,小脑,睾丸和肺脏表达量较高;而肝脏和肌肉未表达。新生小鼠9种脏器中,Cpne5表达量最高的是脑组织,眼和肾次之,肺和肝脏表达量很少。制备并评估Cpne5原杂探针的产量,SP6转录的反义探针浓度为100ng/μL,T7转录的正义探针浓度为10ng/μL,原位杂交结果显示Cpne5 mRNA主要表达于胎鼠的端脑、间脑、中脑及菱脑原节。结论在新生和成年小鼠的脑组织中Cpne5基因mRNA高水平表达,并在胎鼠发育过程中定位于胎脑。Objective To investigate the expression of mouse Cpne5 gene mRNA in various tissues and its loca- tion in embryo. Methods After total RNA of main organs extracted, Cpne5 mRNA were detected by RT-PCR. The probe targeting Cpne5 mRNA was synthesized, and the Cpne5 mRNA expression of different gestational age mice embryos were detected by whole-mount hybridization in situ. Results Cpne5 mRNA was detected in ten kinds of tissues of adult mouse. High expression of Cpne5 mRNA was detected in cerebrum, cerebellum, testis and lung, but no signal in liver and muscle. In neonatal mice, the highest expression of Cpne5 mRNA was found in brains, followed by the tissues of eyes and kidney. The expression level in lung and liver were lower. The concentration of antisense probes transcripted by SP6 promoter was 100ng/μL, and the concentration of sense probes transcripted by T-/ promoter was 10ng/ixL. The hybridization experiments showed that Cpne5 mRNA was mainly expressed in telencephalon, diencephalon, mesencephalon and rhombomere of mouse embryo. Conclusion The mouse Cpne5 mRNA are highly expressed in brain tissues of neonatal and adult mice and located in brain in the process of mouse embryonic development.

关 键 词：Cpne5基因 信使RNA 表达 整体原位杂交 小鼠 

分 类 号：R341[医药卫生—基础医学] Q75[生物学—分子生物学]