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水通道蛋白2在小鼠内耳中的发育表达。

Developmental expression of aquaporin 2 in the mouse inner ear.

作者信息

Merves M, Bobbitt B, Parker K, Kishore B K, Choo D

机构信息

Department of Otolaryngology--Head and Neck Surgery, University of Cincinnati College of Medicine, Ohio, USA.

出版信息

Laryngoscope. 2000 Nov;110(11):1925-30. doi: 10.1097/00005537-200011000-00030.

Abstract

OBJECTIVES

The maintenance of endolymph homeostasis is critical for the inner ear to perform its functions of hearing and maintaining balance. The identification and cloning of aquaporins (a family of water channel proteins) has allowed the study of a novel cellular mechanism potentially involved in endolymph homeostasis. The objective of the present study was to define the developmental temporal and spatial expression pattern of aquaporin 2 (Aqp2) in the developing mouse inner ear.

STUDY DESIGN

A systematic immunohistochemical study of Aqp2 protein expression was performed on embryonic mouse inner ears ranging from embryonic day 10 (otocyst stage) to embryonic day 18 (just before birth).

METHODS

Serial cryosections of embryonic mouse inner ears were used for immunohistochemical experiments. A rabbit polyclonal antisera raised against a synthetic Aqp2 peptide was used with a standard nickel intensified 3,3-diaminobenzidine reaction protocol for immunolocalization of Aqp2 in tissue sections.

RESULTS

Aquaporin 2 is expressed diffusely in the early otocyst, then becomes progressively restricted as the inner ear matures. During early cochlear duct formation (embryonic days 12 and 13), expression of Aqp2 is homogeneous; later, it becomes restricted to specific regions of the endolymphatic compartment (embryonic days 15 and 18). Similar restriction of expression patterns could be noted for the vestibular structures. Endolymphatic duct and sac and stria vascularis expression of Aqp2 was noted to occur fairly late during development but demonstrated a distinct pattern of immunolabeling.

CONCLUSIONS

Aquaporin 2 shows an early and specific pattern of expression in the developing mouse inner ear, suggesting a significant role for this water channel protein in the development of endolymph homeostasis and meriting further functional studies of Aqp2 in the inner ear.

摘要

目的

内淋巴液稳态的维持对于内耳执行其听觉和维持平衡的功能至关重要。水通道蛋白(一类水通道蛋白家族)的鉴定和克隆使得研究一种可能参与内淋巴液稳态的新型细胞机制成为可能。本研究的目的是确定水通道蛋白2(Aqp2)在发育中的小鼠内耳中的发育时间和空间表达模式。

研究设计

对胚胎期第10天(耳泡期)至胚胎期第18天(即将出生)的胚胎小鼠内耳进行了Aqp2蛋白表达的系统免疫组织化学研究。

方法

胚胎小鼠内耳的连续冰冻切片用于免疫组织化学实验。使用针对合成Aqp2肽产生的兔多克隆抗血清,采用标准的镍增强3,3 -二氨基联苯胺反应方案对内耳组织切片中的Aqp2进行免疫定位。

结果

水通道蛋白2在早期耳泡中广泛表达,然后随着内耳成熟逐渐受到限制。在耳蜗管早期形成阶段(胚胎期第12天和第13天),Aqp2的表达是均匀的;后来,它局限于内淋巴腔的特定区域(胚胎期第15天和第18天)。前庭结构也有类似的表达模式限制。内淋巴管和内淋巴囊以及血管纹中Aqp2的表达在发育过程中出现得相当晚,但呈现出明显的免疫标记模式。

结论

水通道蛋白2在发育中的小鼠内耳中呈现早期且特异的表达模式,表明这种水通道蛋白在内淋巴液稳态的发育中起重要作用,值得对内耳中的Aqp2进行进一步的功能研究。

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