Clcn3敲除对幼龄小鼠实质器官的影响

Effect of Clcn3 knockout on parenchymal organs in young mice

目的探究Clcn3敲除对幼龄小鼠实质器官发育的影响,为Clcn3基因敲除动物应用提供理论参考。方法PCR鉴定FVB小鼠基因型:野生型(Clcn3^(+/+))、杂合子(Clcn3^(+/-))及纯合子(Clcn3-/-);小鼠3周龄时,采集心、肝、脾、肺、肾和脑等主要实质器官并拍照,利用Image J软件测量其投影面积;中性福尔马林溶液固定各实质器官,石蜡切片进行HE染色,全切片利用全自动数字病理扫描分析系统扫描后观察器官细微结构。结果与Clcn3^(+/+)和Clcn3^(+/-)小鼠相比,Clcn3-/-小鼠体重显著降低(P<0.05);Clcn3缺失会显著降低小鼠心、肝、脾、肺及肾的体积(P<0.05),但不会影响小鼠脑的大小(P>0.05);HE染色显示,Clcn3-/-小鼠各实质器官组织结构未见异常,实质细胞体积相对较小。结论Clcn3敲除导致幼龄小鼠的实质器官减小,未对幼龄小鼠实质器官产生明显病理损伤。

Objective To explore the effect of Clcn3 gene knockout on parenchymal organs in mice to provide reference data for the application of Clcn3 gene knockout mice.Methods Wild-type(Clcn3^(+/+)),heterozygous(Clcn3^(+/-)),and homozygous mice(Clcn3-/-)were identified by polymerase chain reaction.Heart,liver,spleen,lung,kidney,and brain were collected from 3-week-old mice and photographed,and their projection areas were measured using ImageJ software.The parenchymal organs were then fixed in neutral formalin solution,embedded in paraffin,sectioned,and stained with hematoxylin and eosin(HE).The slides were then scanned using a automated digital pathology scan analysis system to observe changes in tissue structure.Results Body weight was significantly decreased in homozygous knockout mice compared with wild-type and heterozygous mice(P<0.05).Deletion of Clcn3 significantly reduced the volumes of the heart,liver,spleen,lung,and kidney(P<0.05),but not the brain(P>0.05).HE staining indicated that the structures of the parenchymal organs were normal in Clcn3-/-mice,but the parenchymal cell volume was relatively small.Conclusions Knockout of Clcn3 could reduce the sizes of the major parenchymal organs in young mice,but did not cause any obvious pathological damage to these organs.