Regulation of the PTH/PTHrP type-2 receptor and TIP39

描述(申请人提供)：一种G蛋白偶联甲状旁腺激素/甲状旁腺激素受体2型 硬骨鱼和哺乳动物的受体(PTH2 R)与 脊椎动物PTHfPTHrP型受体(PTH1R)。以努力隔离 提纯了一种含39个氨基酸残基的PTH2 R同源配体 来自牛下丘脑(39个残基的漏斗结节多肽， BTIP39)，它选择性和有效地刺激哺乳动物和硬骨鱼 PTH_2R。因此，TIP39似乎是TIP39的内源性配体。 脊椎动物PTH2 R。近年来，人类和小鼠的基因组DNA和cDNA序列 TIP39的分离，编码配体的体外鉴定，以及它们的 多种不同成人神经内分泌的组织特异性表达 纸巾。此外，VTH2 R已被证明具有空间和时间特性 在早期发育的肾脏、脑和睾丸中的表达。到目前为止， 只有几种斑马鱼假定的PTH2 R剪接变异体已被鉴定； 未知的是人类和人类基因组调控元件的鉴定 斑马鱼PTH2Rs以及TIP39。TIP39基因启动子的鉴定 而PTH2 R可以阐明它们的组织特异性表达并提供 关于这一新的生理分支的重要信息 发现了内分泌系统。为了检验TIP39-PTH2 R系统的假设 在不同的内分泌和神经组织中有受调控的表达，这 提案描述了基因和转录本的分子克隆 斑马鱼TIP39，相应启动子的鉴定(特定目的 1)，以及斑马鱼TIP39的功能特性(特定目标2)。在……里面 此外，本方案还将克隆斑马鱼PTH2R基因，以确定 调控元件，控制基因的空间和时间表达 PTH2 R和假定的剪接变异体(具体目标3)。新颖性和 斑马鱼作为动物模型的创新提供了一个强大的模型 研究TIP39和PTH2 R在几种 神经内分泌系统，这可能为它们的生物学提供进一步的线索 在哺乳动物中的角色。因此，这项工作将启动一个长期的分子 TIP39-PTH2R系统作用的内分泌和发育研究 在脊椎动物的早期发育阶段。此外，未来的研究可能会启动 研究TIP39和PTH2R基因调控的分子机制 表情。

DESCRIPTION (provided by applicant): A G-protein coupled PTH/PTHrP type-2 receptor (PTH2R) from teleosts and mammals shows significant homology to the vertebrate PTHfPTHrP type-1 receptor (PTH1R). In an effort to isolate the cognate ligand of the PTH2R, a peptide of 39 amino acid residues was purified from the bovine hypothalamus (tuberoinfundibular peptide of 39 residues, bTIP39), which selectively and potently stimulates the mammalian and teleost PTH2R. It therefore appears that TIP39 is the endogenous ligand of the vertebrate PTH2R. Recently, genomic DNA and cDNA sequences for human and murine TIP39 were isolated, the encoded ligands characterized in vitro, and their tissue specific expression identified in many diverse adult neuroendocrine tissues. In addition, the VTH2R has been shown to have a spatial and temporal expression during early development in the kidney, brain, and testis. To date, only several zebrafish putative PTH2R splice variants have been identified; unknown are the identification of genomic regulatory elements for the human and zebrafish PTH2Rs, as well as TIP39. The identification of promoters for TIP39 and the PTH2R may clarify their tissue specific expression and provide significant information regarding the physiological ramifications of this newly discovered endocrine system. To test the hypothesis that the TIP39-PTH2R system has a regulated expression in diverse endocrine and neuronal tissues, this proposal describes the molecular cloning of the gene and transcript for zebrafish TIP39, identification of the corresponding promoters (Specific Aim 1), and functional characterization of zebrafish TIP39 (Specific Aim 2). In addition, this proposal will clone the zebrafish PTH2R gene to determine the regulatory elements, which control the spatial and temporal expression of the PTH2R and the putative splice variants (Specific Aim 3). The novelty and innovation of using zebrafish as an animal model provides a powerful model system to study the role of TIP39 and the PTH2R in development of several neuro-endocrine systems, which may provide further clues for their biological roles in mammals. Therefore, this work will initiate a long-term molecular endocrine and developmental investigation of the role of the TIP39-PTH2R system in early vertebrate development. Furthermore, future studies may be initiated in characterizing the molecular mechanisms regulating the TIP39 and PTH2R gene expression.