摘要
目的构建海马组织高表达Neuritin的基因小鼠并进行繁殖、基因型鉴定,为研究上调Neuritin表达对2型糖尿病认知功能障碍作用机制提供模型。方法将4对Emx1-cre基因小鼠、Neuritin-Loxp基因小鼠和db/db纯合子小鼠进行配对繁殖,提取小鼠尾巴DNA样本,经PCR法鉴定各繁殖后子代小鼠的基因型,获得杂合子Loxp/Emx1-cre/db基因小鼠,繁殖并鉴定该小鼠子代基因型,获得纯合子loxp/^(+)_Emx1-cre/^(+)_db/db/^(+)基因小鼠。以C57BL/6J野生型小鼠作为正常对照组,采用免疫荧光进一步确定模型的成功性,采用尼氏染色观察各组神经元病理形态。结果3种转基因型小鼠均有较好的繁殖能力,相互杂交其子鼠基因型为loxp/^(+)_Emx1-cre/^(+)、loxp/^(-)_Emx1-cre/^(+)、loxp/^(+)_Emx1-cre/^(-)、loxp/^(+)_Emx1-cre/^(+)_db/db/^(-)、loxp/^(-)_Emx1-cre/^(+)_db/db/^(-)、loxp/^(+)_Emx1-cre/^(-)_db/db/^(-)、loxp/^(+)_Emx1-cre/^(+)_db/db/^(+)、loxp/^(-)_Emx1-cre/^(+)_db/db/^(+)、loxp/^(+)_Emx1-cre/^(-)_db/db/^(+)等,上述基因小鼠繁殖结果基本符合孟德尔遗传定律。免疫荧光结果显示:loxp/^(+)_Emx1-cre/^(+)_db/db/^(+)基因小鼠的Neuritin较Db/db/纯合子基因小鼠表达量较高,差异有统计学意义(F=18.15,P<0.05)。结论PCR鉴定小鼠基因型的方法可靠,loxp/^(-)_Emx1-cre/^(+)基因型小鼠与Db/db纯合子小鼠交配是获得loxp/^(+)_Emx1-cre/^(-)_db/db/^(+)基因型小鼠的有效途径。
Objective To construct,breed,and genetically characterize a genetic mouse model with high Neuritin expression in hippocampus,and to provide a model for studying the mechanism of up-regulation of Neuritin expression on cognitive dysfunction in type 2 diabetes mellitus.Methods Four pairs of Emx1-cre gene mice,Neuritin-Loxp gene mice,and db/db purebred mice were bred in pairs,and mouse tail DNA samples were extracted.The genotypes of the offspring mice of each breeding were identified by PCR to obtain heterozygous Loxp/Emx1-cre/db gene mice,which were bred and genotyped to obtain purebred loxp/^(+)_Emx1-cre/^(+)_db/db/^(+)gene mice.C57BL/6J wild-type mice were used as the normal control group.Immunofluorescence was used to further determine the success of the model,and Nichols staining was used to observe the neuronal pathology and morphology in each group.Results All three transgenic mice had good reproductive ability,and the genotypes of their offspring crossed with each other were loxp/^(+)_Emx1-cre/^(+),loxp/^(-)_Emx1-cre/^(+),loxp/^(+)_Emx1-cre/^(-),loxp/^(+)_Emx1-cre/^(+)_db/db/^(-),loxp/^(-)_Emx1-cre/^(+)_db/db/^(-),loxp/^(+)_Emx1-cre/^(-)_db/db/^(-),loxp/^(+)_Emx1-cre/^(+)_db/db/^(+),loxp/^(-)_Emx1-cre/^(+)_db/db/^(+),and loxp/^(+)_Emx1-cre/^(-)_db/db/^(+),etc.The results of the above genes of mouse reproduction are basically consistent with Mendelian.The breeding results of mice with the above genes basically conformed to Mendel's law of inheritance.Immunofluorescence results showed that loxp/^(+)_Emx1-cre/^(+)_db/db/^(+)mice had higher expression of Neuritin than Db/db/pureblood mice,and the difference was statistically significant(F=18.15,P<0.05).Conclusion PCR is a reliable method for identifying mouse genotypes,and mating loxp/^(-)_Emx1-cre/^(+)genotyped mice with Db/db purebred mice is an effective way to obtain loxp/^(+)_Emx1-cre/^(-)_db/db/^(+)genotyped mice.
作者
李和教
卢伟锋
LI He-jiao;LU Wei-feng(School of Chemical and Materials Engineering,Hainan University of Science and Technology,Haikou 571126,Hainan,CHINA;Department of Pharmacy,the First Affiliated Hospital of Hainan Medical University,Haikou 570102,Hainan,CHINA;Cancer Institute,the First Affiliated Hospital of Hainan Medical University,Haikou 570102,Hainan,CHINA)
出处
《海南医学》
CAS
2024年第19期2737-2742,共6页
Hainan Medical Journal
基金
2021年海南省自然科学基金(编号:821QN402)
2023年海南省自然科学基金(编号:823QN347)
海南医学院第一附属医院2020年度青年培育基金项目(编号:HYYFYPY202001)
海南医学院2020年科研培育基金(编号:HYPY2020035)。
