Merves Michele, Krane Carissa M, Dou Hongwei, Greinwald John H, Menon Anil G, Choo Daniel
Department of Pediatric Otolaryngology, Center for Hearing and Deafness Research, Cincinnati, OH 45229, USA.
J Assoc Res Otolaryngol. 2003 Jun;4(2):264-75. doi: 10.1007/s10162-002-3033-7.
To examine the potential roles of aquaporins 1 and 5 (AQP1 and AQP5, respectively) in inner ear development and function, we defined their spatial and temporal expression patterns in the developing mouse inner ear and examined the morphologic and physiologic effects of loss of Aqp5 function. Standard in situ hybridization (ISH) and immunohistochemical (IHC) assays were used for expression studies with routine morphologic, behavioral, and physiologic assessments of hearing and balance in Aqp5 null mutant mice. AQP1 was first detected at embryonic day 10.5 (E10.5) in the otocyst but eventually localized to specific nonsensory portions of the inner ear and connective tissue cells surrounding the membranous labyrinth. AQP5 displayed specific cochlear expression, first detectable at E15.5 in the nonsensory epithelium and later restricted to the lateral wall of the cochlear duct near the spiral prominence. AQP5 expression continued through postnatal periods with a change of expression domain to the stria vascularis between postnatal day 7 (P7) and P14. By in situ hybridization and immunohistochemical techniques, subtle differences between transcript and protein expression patterns were noted for both AQP1 and 5. Although AQP5 is dynamically expressed in the developing mouse inner ear, adult Aqp5 knockout mice show normal hearing when tested and normal inner ear structural development. These results suggest redundant or alternative mechanisms that likely regulate water homeostasis in the developing and mature inner ear.
为了研究水通道蛋白1和5(分别为AQP1和AQP5)在内耳发育和功能中的潜在作用,我们确定了它们在发育中的小鼠内耳中的时空表达模式,并研究了Aqp5功能缺失的形态学和生理学影响。使用标准原位杂交(ISH)和免疫组织化学(IHC)分析进行表达研究,并对Aqp5基因敲除小鼠的听力和平衡进行常规形态学、行为学和生理学评估。AQP1最早在胚胎第10.5天(E10.5)在耳囊中被检测到,但最终定位于内耳的特定非感觉部分以及膜迷路周围的结缔组织细胞。AQP5表现出特定的耳蜗表达,最早在E15.5时在非感觉上皮中可检测到,随后局限于靠近螺旋隆起的耳蜗管侧壁。AQP5的表达在出生后阶段持续存在,在出生后第7天(P7)至P14之间表达域发生变化,变为血管纹。通过原位杂交和免疫组织化学技术,发现AQP1和5在转录本和蛋白质表达模式上存在细微差异。尽管AQP5在发育中的小鼠内耳中动态表达,但成年Aqp5基因敲除小鼠在测试时听力正常,内耳结构发育也正常。这些结果表明,可能存在冗余或替代机制来调节发育中和成熟内耳中的水平衡。