Prkra小耳小鼠动物模型在先天性小耳畸形研究中的应用  

Application of Prkra^lear-3J mouse as an animal model in the study of congenital microtia

作　　者：刘伟 林琳 杨庆华 潘博 何乐人 蒋海越 Liu Wei;Lin Lin;Yang Qinghua;Pan Bo;He Leren;Jiang Haiyue(First Center of the External Ear Plasty and Reconstruction,Plastic Surgery Hospital,Chinese Academy of Medical Science&Peking Union Medical College,Beijing 100144,China)

机构地区：[1]中国医学科学院北京协和医学院整形外科医院耳整形再造一中心,100144

出　　处：《中华整形外科杂志》2019年第8期804-808,共5页Chinese Journal of Plastic Surgery

基　　金：北京市自然科学基金(7132176);首都临床特色应用研究(Z161100000516098);中国医学科学院医学与健康科技创新工程项目(2016-I2M-2-006,2017-I2M-1-007,2016-I2M-2-001);中国医学科学院颅面先天性畸形发病机制研究重点实验室项目(2018PT31051);协和青年基金(3332016133)

摘　　要：目的研究Prkra小耳小鼠动物模型的构建方法,并探讨其用于先天性小耳畸形研究的可行性。方法从美国JAX实验室引进同窝Prkra little ear(小耳)小鼠6只,通过繁育共获得13只Prkra小耳小鼠孕鼠。其中10只孕鼠共取75只胎龄为14.5 d的胎鼠尾,提取DNA进行Sanger法测序;3只孕鼠正常生产,幼鼠出生后常规饲养,取耳廓畸形和耳廓正常的40 d子鼠各3只,进行骨骼阿利新蓝染色,并作对比分析。结果Prkra小耳纯合型小鼠耳廓明显比其同胞正常耳廓小,失去了正常小鼠耳廓的亚结构形态。DNA测序结果表明,突变位点为在2号染色体76、643、218 bp(GRCm38)上的G向A突变,该突变位点位于第3外显子之后。阿利新蓝染色结果显示,Prkra小耳纯合型小鼠体型较正常野生型小,四肢骨发育基本正常,但较细短,后肢表现尤为明显,肋骨发育较细小,尾骨发育较细短,头颅发育略小。结论成功构建了Prkra小耳小鼠模型,而且该模型是研究先天性小耳畸形胚胎发育机制比较合适的动物模型。Objective To explore the feasibility of using Prkra^lear-3J mouse(also known as:little ears 3 Jackson)as an animal model in the study of congenital microtia.Methods Six Prkra^lear-3J mice of the same litter were introduced from JAX laboratory in the United States.13 pregnant Prkra^lear-3J mice were obtained through breeding.A total of 75 fetus tails with gestational age of 14.5 days were obtained,and DNA was extracted and sequenced by Sanger method.Three pregnant mice were normally produced,and the young mice were routinely reared after birth.Three 40-day-old mice with auricle deformity and normal auricle were used respectively to perform skeletal alcian blue staining and comparative analysis.Results The auricle of Prkra^lear-3J homozygous mice was significantly smaller than that of the wild-type mice,and the substructure morphology of the normal mouse auricle was lost.The mutation site was a G-to-A mutation on chromosome 76,643,218 bp(GRCm38),which was located after the third exon.The results of alcian blue staining showed that the overall body size of Prkra^lear-3J homozygous mouse was smaller than that of wild type mouse,and the bone development of limbs was basically normal,but it was thin and short,especially in the hind limbs,the ribs were smaller,the tailbone was shorter,and the skull was slightly smaller.Conclusions The Prkra^lear-3J mouse is a suitable animal model for studying the developmental mechanism of congenital microtia.

关 键 词：Prkra小耳小鼠 小耳畸形 动物模型 Prkra PACT 阿利新蓝染色 

分 类 号：R28[医药卫生—中药学]