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作 者:井然[1] 谢启应[1] 李非[1] 邓桂元[1] 龙添翼 邹隽琳 欧阳昆富 JING Ran, XIE Qiying, LI Fei, DENG Guiyuan, LONG Tianyi, ZOU Juanlin, OUYANG Kunfu(Cardiovascular Department of Xiangya Hospital, Central South University, Changsha 410004, Hunan, China)
机构地区:[1]中南大学湘雅医院心内科
出 处:《中国临床药理学与治疗学》2018年第8期862-866,共5页Chinese Journal of Clinical Pharmacology and Therapeutics
基 金:国家自然科学基金青年科学基金项目(81700289)
摘 要:目的:探讨蛋白激酶D1(protein kinase D1,PKD1)与蛋白激酶D3(protein kinase D3,PKD3)基因在小鼠胚胎发育早期心脏组织中的表达及意义。方法:构建PKD1与PKD3双敲除基因小鼠模型,RNA原位杂交实验检测PKD1与PKD3在胚胎发育E9.5与E10.5 d时心脏中的分布情况,Western blot检测PKD1与PKD3蛋白的表达,体视显微镜及HE染色观察心脏的形态学及组织学。结果:E9.5 d时,PKD1与PKD3在心脏流出道、右心房、右心室等位置均可见表达。在E10.5d,PKD1与PKD3在右心房均高表达,在流出道也可见表达。在E9.5 d时,PKD1与PKD3在心脏的外型上没有明显差异;在E10.5 d时,所有双敲除小鼠都有明显的胸腔积水与心腔扩大的表型,心壁很薄;从E12.5 d开始就见不到双敲除小鼠。E8.5 d与E9.5 d时,双敲除小鼠的基因型分布符合孟德尔定律。结论:PKD1和PKD3均参与小鼠胚胎的心脏发育,但是其具体作用机制及相互之间的关系仍需进一步研究。AIM: To explore the expression of PKD1 and PKD3 genes in heart tissue during early embryonic development in mice.METHODS: To construct the PKD1 and PKD3 double knockout mice model,the RNA in situ hybridization test was used to detect the distribution of PKD1 and PKD3 in the cardiac development of E9.5 and E10.5.Protein expression of PKD1 and PKD3 was detected by western blot,and the cardiac morphology and histology were observed by stereoscopy and HE staining.RESULTS: At E9.5,PKD1 and PKD3 were expressed in the outflow tract,right atrium and right ventricle.At E10.5,PKD1 and PKD3 were highly expressed in the right atrium,and are also expressed in outflow tract.At E9.5,there was no significant difference in the heart form between PKD1 and PKD3; at E10.5,all double knockout mice had obvious pleural effusion and enlargement of the heart cavity,and the wall of the heart was thin; the double knockout mice were not seen from E12.5.At E8.5 and E9.5, the genotype distribution of the doubleknockout mice conforms to Mendel 's law.CONCLUSION: Both PKD1 and PKD3 are involved in the heart development of mouse embryos,but the specific mechanism of action and the relationship between them still need to be further studied.
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