Bmp5短耳小鼠动物模型在先天性小耳畸形研究中的应用

Application of Bmp5 short ear mouse model in the study of congenital microtia

目的研究Bmp5短耳纯合型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Bmp5短耳小鼠12只,通过繁育获得Bmp5短耳小鼠13只孕鼠,10只孕鼠共取86只胎龄在14.5 d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Bmp5短耳纯合型小鼠耳廓明显比其同胞正常耳廓短小,Sanger法测序后发现Bmp5点突变位置在外显子2倒数第7个碱基,碱基由C突变为T。阿利新蓝染色结果可见,纯合型突变畸形小鼠体形较小,胸骨和肋骨发育明显异常。结论Bmp5短耳小鼠模型可以完全适用于人类先天性小耳畸形的研究。

Objective To study the construction of Bmp5 short ear homozygous mouse model and the feasibility of its application in the study of congenital microtia.Methods Twelve Bmp5 short-ear mouse were introduced from JAX laboratory in the United States.Ten pregnant Bmp5 short-ear mouse were obtained by breeding.DNA was extracted from the tail of 86 fetal rats aged 14.5 days and sequenced by Sanger method.Three pregnant mice were born normally,and the young mouse was fed conventionally.Three 40-day-old mice with auricle deformity and normal auricle were stained with Alcian blue,and the result were compared and analyzed.Results The auricle of Bmp5 short ear homozygous mouse was significantly shorter than that of their sibling normal auricles.Sanger sequencing showed that the point mutation of Bmp5 was the seventh base at the bottom of exon 2,and the base mutated from C to T.The result of Alcianblue staining showed that the size of homozygous mutant mouse was small and the development of sternum and ribs was obviously abnormal.Conclusions The model of Bmp5 short ear mouse can be completely suitable for the study of human congenital microtia.The model of Bmp5 short ear mouse is a suitable animal model to study the development mechanism of congenital microtia.