Functional role of transporter Slc4a11 (BTR1/NaBC1) in the cornea

转运蛋白 Slc4a11 (BTR1/NaBC1) 在角膜中的功能作用

基本信息

批准号：
8243356
负责人：
MICHAEL L JENNINGS
金额：
$ 23.66万
依托单位：
UNIV OF ARKANSAS FOR MED SCIS
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-12-01 至 2013-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8243356
关键词：
Animal Model Antibodies Apoptosis Aqueous Humor Bicarbonates Bilateral Borates Boron Carrier Proteins Cattle Cell Line Cells Clinical assessments Confocal Microscopy Cornea Corneal Endothelium Corneal Stroma Corneal dystrophy Coupled Data Diffuse Edema Endoplasmic Reticulum Endothelial Cells Exhibits Face Family Fuchs&apos Endothelial Dystrophy Functional disorder Genes Genetic Goals Guidelines Hearing Hereditary Disease Human Hydration status Immunohistochemistry Individual Inherited Ion Transport Ions Knock-in Mouse Knockout Mice Knowledge Label Link Liquid substance Location Maintenance Mass Spectrum Analysis Measurement Measures Mediating Microelectrodes Microscope Molecular Monitor Mus Mutation Phenotype Physiological Plasma Play Point Mutation Positioning Attribute Process Protein Family Proteins Publishing Reporting Role Symptoms Syndrome Transgenic Mice Transgenic Organisms Vision Xenopus oocyte alkalinity apical membrane aqueous base basolateral membrane extracellular fascinate man member mouse model mutant overexpression research clinical testing research study solute

项目摘要

DESCRIPTION (provided by applicant): Congenital hereditary endothelial dystrophy (CHED) causes impaired vision resulting from opacification of the cornea. The recessive form of CHED (CHED2) as well as some types of Fuch's endothelial dystrophy and Harboyan syndrome were recently shown to be associated with mutations in the SLC4A11 gene that encodes BTR1/NaBC1, a member of the bicarbonate transporter family. The function and subcellular location of this protein in cornea are unknown, and there is currently no adequate animal model for the study of CHED caused by SLC4A11 mutations. The goals of the proposed experiments are to determine the molecular function of SLC4A11 and its localization in the cornea, and to generate a mouse model for CHED2. The available functional information suggests that SLC4A11 transports Na+ and borate [B(OH)4- ], although no boron transport experiments with this protein have been performed. Specific Aim 1A is to use HEK293 cells and a corneal endothelial cell line, with mass-spectrometry, to determine whether mammalian (mouse, human) SLC4A11 cotransports Na+ and boron. Specific Aim 1B is to use Xenopus oocytes, with ion-selective microelectrodes, to determine whether SLC4A11 cotransports Na+ and HCO3-. In Specific Aim 2 we will perform immunohistochemistry to determine whether Slc4a11 is located in the basolateral or apical membrane of the mouse corneal endothelium. In Specific Aim 3 we will generate and evaluate the phenotype of a mouse model of CHED2 prepared by introducing, to mouse Slc4a11, a point mutation (in the position of human R755Q) that is known to cause CHED2 in humans. PUBLIC HEALTH RELEVANCE: Maintenance of transparency of the cornea is essential for good vision. A variety of conditions can cause the cornea to become cloudy, including an inherited condition known as congenital hereditary endothelial dystrophy (CHED). Some forms of CHED are a result of mutations in a transport protein known as SLC4A11, the function of which has not been well established. The goals of this project are to determine the function of SLC4A11 and to establish a mouse model of CHED.

描述（由申请人提供）：先天性遗传性内皮营养不良（CHED）会因角膜浑浊而导致视力受损。最近显示，隐性形式的 CHED (CHED2) 以及某些类型的 Fuch 内皮营养不良和 Harboyan 综合征与编码 BTR1/NaBC1（碳酸氢盐转运蛋白家族成员）的 SLC4A11 基因突变有关。该蛋白在角膜中的功能和亚细胞位置尚不清楚，目前还没有足够的动物模型来研究 SLC4A11 突变引起的 CHED。拟议实验的目标是确定 SLC4A11 的分子功能及其在角膜中的定位，并生成 CHED2 的小鼠模型。现有的功能信息表明，SLC4A11 转运 Na+ 和硼酸盐 [B(OH)4- ]，尽管尚未对该蛋白进行硼转运实验。具体目标 1A 是使用 HEK293 细胞和角膜内皮细胞系，通过质谱分析确定哺乳动物（小鼠、人）SLC4A11 是否共同转运 Na+ 和硼。具体目标 1B 是使用爪蟾卵母细胞和离子选择性微电极来确定 SLC4A11 是否共同转运 Na+ 和 HCO3-。在具体目标 2 中，我们将进行免疫组织化学以确定 Slc4a11 是否位于小鼠角膜内皮的基底外侧膜或顶膜中。在具体目标 3 中，我们将生成并评估 CHED2 小鼠模型的表型，该模型是通过向小鼠 Slc4a11 引入已知会在人类中引起 CHED2 的点突变（在人 R755Q 的位置）而制备的。公众健康相关性：保持角膜透明度对于良好视力至关重要。有多种情况会导致角膜变得浑浊，其中包括一种称为先天性遗传性内皮营养不良 (CHED) 的遗传性疾病。某些形式的 CHED 是一种名为 SLC4A11 的转运蛋白突变的结果，该蛋白的功能尚未明确。该项目的目标是确定SLC4A11的功能并建立CHED小鼠模型。