EVALUATION OF TULP1 IN THE RETINA

拟议研究的总体目标是获取信息， 关于TULP1在视网膜中的作用。 申请人最近 获得的证据表明，TULP1基因是常染色体 隐性视网膜色素变性（ARRP）。这种基因的蛋白质产物 在视网膜中特异性表达， 关于这种蛋白质的功能的信息是存在的。 RP是 遗传性致盲疾病折磨着成千上万的人 的 只有大约20%的RP病例的遗传病因是已知的 而引起疾病的生化途径就更少了。 此应用程序的第一个特定目标集中于本地化TULP1 在视网膜上。 将开始确定表达TULP1的细胞类型 来提供蛋白质功能的线索。设计的实验 为了实现目标，将使用分子技术进行 包括原位杂交和免疫组织化学。 第二 目的是分析一种缺乏功能性 TULP1基因。 评估这些小鼠的视网膜将是重要的 可能与这些发现相关的病理变化 在有TULP1突变的人身上。 最后一个具体目标是证明 TULP1基因缺陷导致视网膜变性。 这将通过尝试拯救TULP1敲除小鼠来实现 （假设小鼠退化）通过重新引入野生型基因， 通过产生转基因小鼠，其表达含有已知的 无效背景上的TULP1突变。 评价所有小鼠的方法 包括眼底镜、光学和电子显微镜， 视网膜电图 发现有关功能的信息 TULP1有望提供有关参与的途径的知识， 视网膜变性

The overall objective of the proposed studies is to obtain information regarding the role of TULP1 in the retina. The applicant recently obtained evidence pointing to the TULP1 gene as a cause for autosomal recessive retinitis pigmentosa (ARRP). The protein product of this gene is specifically expressed in the retina, but little additional information regarding the function of this protein exists. RP is a hereditary blinding disorder afflicting thousands of people. The genetic etiology of RP is known for only about 20 percent of the cases and the biochemical pathways involved in causing the disease even less. The first specific aim of this application focuses on localizing TULP1 in the retina. Determining the cell type(s) expressing TULP1 will begin to give clues as to the function of the protein. Experiments designed to achieve aim one will be conducted using molecular techniques including in situ hybridization and immunohistochemistry. The second aim involves analyzing a transgenic mouse strain lacking a functional TULP1 gene. It will be important to evaluate the retina of these mice for possible pathologic changes that might correlate with those found in humans with TULP1 mutations. The last specific aim involves proving that identified defects in the TULP1 gene cause retinal degeneration. This will be done by attempting to rescue the TULP1 knock-out mice (assuming the mice degenerate) by re-introducing the wild-type gene and by generating transgenic mice expressing constructs containing known TULP1 mutations on the null background. Methods to evaluate all mice include funduscopy, light and electron microscopy, and electroretinography. Discovering information regarding the function of TULP1 will hopefully provide knowledge about pathways involved in retinal degeneration.