小鼠内耳发育过程中C1ql1基因的表达特点

Expression of C1ql1 Gene during of Inner Ear Development in Mice

目的 探究C1ql1基因在不同发育阶段小鼠耳蜗细胞中的表达特点及在内耳发育中的作用。方法 选取出生后1 d(P1)、7 d(P7)、14 d(P14)、28 d(P28)、56 d(P56)的KM小鼠各6只进行耳蜗基底膜铺片、耳蜗中轴冷冻切片及免疫荧光染色,采用共聚焦显微镜观察不同发育阶段小鼠耳蜗中C1ql1基因的表达特点,通过逆转录聚合酶链反应、Western blot检测不同发育阶段小鼠耳蜗中C1ql1基因的表达量变化。结果 发育期小鼠耳蜗螺旋神经节细胞、毛细胞中C1ql1基因表达量随年龄增长逐渐增加。P1小鼠耳蜗无C1QL1蛋白条带,P7、P14、P28、P56小鼠耳蜗C1QL1蛋白条带均可清晰显影。P1小鼠C1QL1蛋白未明确表达,P7平均光密度值与P14、P28、P56比较,差异均有统计学意义(P<0.05)。P14、P28、P56平均光密度值比较,差异均无统计学意义(P>0.05)。C1ql1基因在听觉发育关键期小鼠耳蜗听神经纤维有一过性表达,出生后7 d表达最高,成年后消失。结论 C1ql1基因在小鼠耳发育和听觉功能形成过程中起着重要作用。

Objective To report expression characteristics and significance of the C1ql1 gene in mouse cochlear cells during different developmental stages.Methods Kunming mice at 1 day,7 days,14 days,28 days and 56 days of age(n=6 each)were selected,and expression of the C1ql1 gene on the basilar membrane and in the cochlea was examined by confocal microscopy with immunofluorescence,as well as RT-PCR and Western blot.Results The level of C1ql1 gene expression in spiral ganglion cells and hair cells gradually increased with age.C1QL1 protein bands were lacking in P1 mice,but clearly visualized in P7,P14,P28 and P56 mice,with average optical density of the C1QL1 protein in P7 mice showing a difference as compared with that in P14,P28 and P56 mice(P<0.05)and no statistically significant difference among the latter three age groups(P>0.05).The C1ql1 gene was transiently expressed in auditory nerve fibers during the critical period of auditory development,peaking on the 7th day after birth while absent in adult mice.Conclusion The C1ql1 gene play an important role in inner ear development and the formation of auditory function.