Role of Aquaporin in vocal fold epithelial water fluxes.

描述(由申请人提供)：这项建议的长期目标是更好地了解细胞机制，它调节声带中的跨上皮水流量。我们推测，存在于声带上皮细胞质膜上的水通道调节声带的渗透跨上皮水通量。如果是这样的话，水通道蛋白可能为调节声带水合和发声提供了一种内在的机制。我们将首先通过免疫印迹分析证明天然声带上皮组织中存在两个水通道蛋白，然后使用免疫金标记法和透射电子显微镜对水通道蛋白进行免疫定位。水通道蛋白在声带跨上皮水通量中的作用将通过对管腔上皮施加渗透梯度来阐明。水通道蛋白的表达依赖于渗透压梯度，将通过Northern Blot分析水通道蛋白的mRNA表达并测量相关的跨上皮水通量来证实。这里描述的数据将有助于了解水通过生物膜的通量以及层状鳞状上皮组织的细胞生物学，并可能有助于语音障碍临床治疗的进展。

DESCRIPTION (provided by applicant): The long-term goal of this proposal is to better understand the cellular mechanisms, which regulate transepithelial water flux in the vocal fold. We hypothesize that aquaporin water channels, present in the plasma membrane of vocal fold epithelial cells, regulate osmostic transepithelial water fluxes of the vocal fold. If so, aquaporins may provide an intrinsic mechanism for regulation of vocal fold hydration and phonation. We will first demonstrate the presence of two aquaporin water channels in native vocal fold epithelial tissue via Western immunoblot analysis, followed by immunolocalization of the water channel proteins using immunogold labeling and transmission electron microscopy. The role of aquaporins in vocal fold transepithelial water fluxes will then be elucidated by imposition of osmotic gradients to the luminal epithelium. The dependence of aquaporin expression on the osmotic gradient will be demonstrated by Northern Blot analysis of mRNA expression of aquaporins and measuring associated transepithelial water fluxes. Data described here will contribute to knowledge of the flux of water across biological membranes as well as the cell biology of stratified squamous epithelial tissue, and may contribute to advances in clinical treatment of voice disorders.