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

PTH/PTHrP 2 型受体和 TIP39 的调节

• 批准号: 6504614
• 负责人: DAVID Alan RUBIN
• 金额: $ 12.5万
• 依托单位: ILLINOIS STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-01 至 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6504614
• 关键词: early embryonic stage; gene expression; genetic library; genetic regulatory element; hormone receptor; in situ hybridization; molecular cloning; nucleic acid sequence; parathyroid hormone related protein; peptide hormone; polymerase chain reaction; protein structure function; southern blotting; vertebrate embryology; zebrafish

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.

描述（由申请人提供）：G 蛋白偶联 PTH/PTHrP 2 型 来自硬骨鱼和哺乳动物的受体（PTH2R）与 脊椎动物 PTHfPTHrP 1 型受体 (PTH1R)。为了努力隔离 PTH2R 的同源配体，39 个氨基酸残基的肽被纯化 来自牛下丘脑（39 个残基的结节漏斗肽， bTIP39)，选择性地、有效地刺激哺乳动物和硬骨鱼 PTH2R。因此，TIP39 似乎是 TIP39 的内源配体。 脊椎动物 PTH2R。最近，人类和小鼠的基因组 DNA 和 cDNA 序列 TIP39 被分离出来，编码的配体在体外进行了表征，并且它们的 在许多不同的成人神经内分泌中鉴定出组织特异性表达 组织。此外，VTH2R 已被证明具有空间和时间 在肾脏、大脑和睾丸的早期发育过程中表达。迄今为止， 仅鉴定了几种斑马鱼假定的 PTH2R 剪接变体； 人类基因组调控元件的鉴定尚不清楚 斑马鱼 PTH2R 以及 TIP39。 TIP39启动子的鉴定 PTH2R 可以阐明其组织特异性表达并提供 有关这一新发现的生理影响的重要信息 发现内分泌系统。检验 TIP39-PTH2R 系统的假设 在多种内分泌和神经元组织中具有调节表达，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（具体目标 1) 和斑马鱼 TIP39 的功能表征（具体目标 2）。在 此外，该提案将克隆斑马鱼PTH2R基因以确定 调控元件，控制空间和时间的表达 PTH2R 和假定的剪接变体（具体目标 3）。新颖性和 使用斑马鱼作为动物模型的创新提供了一个强大的模型 系统研究 TIP39 和 PTH2R 在几种发育中的作用 神经内分泌系统，这可能为其生物学提供进一步的线索 在哺乳动物中的作用。因此，这项工作将启动长期分子 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过程中。此外，未来的研究可能会启动 表征调节 TIP39 和 PTH2R 基因的分子机制 表达。

项目成果

Bone-remodeling transcript levels are independent of perching in end-of-lay white leghorn chickens.

• DOI: 10.3390/ijms16022663
• 发表时间: 2015-01-23
• 期刊: International journal of molecular sciences
• 影响因子: 5.6
• 作者: Dale MD;Mortimer EM;Kolli S;Achramowicz E;Borchert G;Juliano SA;Halkyard S;Seitz N;Gatto C;Hester PY;Rubin DA
• 通讯作者: Rubin DA