小鼠内耳水通道蛋白的定位及其意义

The orientation and its significance of water channel protein in the mouse inner ear

目的 检测水通道蛋白 (waterchannelprotein ;aquaporin ,Aqp)亚型在小鼠内耳不同部位的分布和各亚型所在的部位。方法 使用成年小鼠 30只 ,经活体灌注 ,切取双侧颞骨 ,按石蜡包埋技术处理和切片。使用免疫组织化学方法标记确认小鼠内耳组织中水通道蛋白亚型 1、3、4、5、7、9分布情况。结果 Aqp 1为 1∶2 0 0和Aqp 3、7、9为 1∶10 0的一抗浓度可以看到它们在内耳不同部位稳定、清晰的染色反应 ,但Aqp 4和 5使用 1∶5 0甚至 1∶30也未观察到染色。Aqp 1在圆窗膜、螺旋韧带、内淋巴囊和内淋巴管、椭圆囊和球囊以及内耳血管壁等处被标记 ;Aqp 3在螺旋韧带、前庭唇、内、外螺旋沟、基底膜和基底膜嵴、内淋巴囊和内淋巴管、膜半规管和椭圆囊、球囊斑等处显示荧光染色。Aqp 7在血管纹、基底膜、前庭膜、椭圆囊和球囊及其囊斑有反应染色 ,而Aqp 9在螺旋缘、前庭唇、内、外螺旋沟、前庭膜、膜半规管和球囊及其囊斑有较强染色。结论 水通道蛋白亚型 1、3、7、9广泛分布于小鼠内耳不同组织中 ,其分布部位和反应强弱存在差异 。

Objective To examine the distribution of various subtypes of water channel protein (aquaporin, Aqp) in various structures of the mouse inner ear. Methods Thirty mice with white color were used in this study and were cardiacally perfused. The temporal bones were taken out and were processed and sectioned by paraffin embedded technique. The sections were labeled with fluorescent antibody by immunohistochemical method. The distribution of Aqp 1, 3, 4, 5, 7, 9 were confirmed in the inner ear of mouse. Results The constant and clear fluorescent reaction could be observed in the inner ear tissue with the first antibody concentration of 1∶200 for Aqp 1 and 1∶100 for Aqp 3, 7, 9. But the reaction of Aqp 4 and 5 could not be found with the concentration of 1∶50, even 1∶30. Aqp 1 was labeled in the round window membrane, spiral ligament, endolymphatic sac and duct, utricle and saccule and the wall of inner ear blood vessel; Aqp 3 in the spiral limb, vestibular lip, internal and external sulci, basilar membrane and basilar membrane crest, endolymphatic sac and duct, membranous semicircular canal and utricle and saccular macula; Aqp 7 in the stria vascularis, basilar membrane, Reissner′s membrane, utricle and saccule and their maculae; Aqp 9 in the spiral limb, vestibular lip internal and external sulci, Reissner′s membrane, membranous semicircular canal, saccule and its macula. Conclusion Aqp 1, 3, 7, 9 were extensively distributed in various tissues of mouse inner ear. Their distribution sites and reaction degree were different, and mainly located in the structures related to the endolymph.