心脏特异表达人源FAM55A转基因小鼠的建立

目的建立心脏特异表达的人源FAM55A转基因小鼠,为研究该基因在心肌病发病中的作用提供模型。方法 Western blot检测FAM55A在野生型小鼠与cTnTR141W转基因小鼠心脏组织中的表达变化及其在野生小鼠的组织表达谱。克隆人源FAM55A基因入α-MHC启动子下游构建a-MHC-FAM55A表达载体,显微注射法建立FAM55A转基因小鼠。PCR鉴定转基因首建鼠的基因型。Western blot鉴定人源FAM55A在转基因小鼠心脏中的表达,超声检测转基因小鼠心脏的几何构型和功能。HE染色检测转基因小鼠心脏的病理改变。结果 FAM55A在野生型小鼠心脏中有少量表达,在扩张型心肌病小鼠的心脏中表达增加。建立了1个心脏组织特异表达人源FAM55A转基因小鼠品系。与野生型小鼠相比,FAM55A转基因小鼠的心脏收缩期和舒张期左室前壁从1月龄到5月龄持续增厚,3月龄转基因小鼠心脏射血分数和短轴缩短率稍有增强,1月龄和5月龄转基因小鼠心脏功能则与同龄野生型小鼠相比无变化。组织学检测显示,转基因小鼠心脏左室心肌细胞不均匀肥大,但不发生紊乱。结论 FAM55A在扩张型心肌病小鼠的心脏中表达上调,建立了心脏特异表达的人源FAM55A转基因小鼠,为进一步和心肌病小鼠模型杂交,研究该基因在心肌病发病中的作用提供了工具。

The Establishment of Cardiac-specific Human FAM55A Transgenic Mice

Objectives To generate the cardiac-specific human FAM55A transgenic mice,a model for the study of its function and effects on cardiomyopathy.Methods The expression of FAM55A gene in the heart tissues from wild type mice and cTnTR141W transgenic mice were analyzed by western blot.The expression of FAM55A in seven different organs was also detected by western blot.The transgenic vector was constructed by inserting the human FAM55A gene into the downstream of α-MHC promoter.The transgenic mice were created by the method of microinjection.The genotype of transgenic line was identified by PCR and the expression level of the gene was determined by Western blot.The pathologic changes of the heart structure and function were analyzed by echocardiography and microscopy.Results The expression of FAM55A was up-regulated in the mice with DCM.One line of C57BL/6J transgenic mice with high human FAM55A expression was identified from five transgenic founders.Compared with the wild type mice,FAM55A transgenic mice showed increased left ventricular anterior wall during systolic and left ventricular anterior wall during diastole from one month of age to five months of age.The cardiac function of transgenic mice was increased at three months of age.The cardiomyocytes from transgenic mice exhibited hypertrophy and no disarrange under light microscope.Conclusions The expression of FAM55A was up-regulated in the cTnTR141W transgenic mice.The transgenic mice with cardiac-specific expression of the human FAM55A gene were generated and it can be used to crossbreed with the DCM model to investigate the effect of FAM55A gene on the development of cardiomyopathy.