Altered expression of PTH type-2 receptor and TIP39

描述（由申请人提供）：在斑马鱼和小鼠大脑中，tuberininfundibular peptide 39 （TIP39）和甲状旁腺激素（PTH） 2型受体（PTH2R）构成了一种神经元mRNA表达保守的新型内分泌系统。与小鼠TIP39-PTH2R系统相关的功能包括痛觉和调节下丘脑（GnRH）和垂体激素（ACTH， GH和LH）。斑马鱼研究表明，TIP39和PTH2R在大脑发育早期与sonic hedgehog基因（Shh）的时空关系中表达。此外，被morpholino抑制TIR39表达的胚胎不能存活，并表现出严重的脑发育障碍。因此，斑马鱼TIP39-PTH2R研究表明，该系统在神经内分泌发育中起着至关重要的作用，可能在胰腺和心脏等其他器官中也有TIR39和PTH2R的表达。因此，斑马鱼（Danio rerio）为研究中枢神经系统（CNS）发育过程中tir39 - pth2r调控机制提供了强有力的模型系统。因此，我们提出了分子分析来更好地阐明斑马鱼PTH2R和TIR39转录物表达改变在早期大脑发育中的影响。1)通过检测Shh、GH、ACTH、LH和GnRH的表达来评估注射mo的胚胎在早期脑发育过程中的PTH2R和TIP39的表达。2)通过检测Shh或Shh信号（sonic you和slow-muscle-省略）缺失的胚胎中表达TIP39的细胞位置，确定大脑发育过程中TIP39和Shh之间的关系。3)通过检测上述标记基因的表达，评估大脑发育早期大脑和下丘脑中PTH2R和TIP39的过表达。由于PTH2R尚未在任何生物体中通过基因敲除进行分析，这些实验将为这种新的TIP39-PTH2R系统在正常和受干扰胚胎中的作用提供新的见解。这项工作是对TIP39-PTH2R系统在中枢神经系统和其他器官发育过程中机制作用的长期分子发育遗传学研究的一部分。

DESCRIPTION (provided by applicant): Tuberoinfundibular peptide 39 (TIP39) and Parathyroid Hormone (PTH) type-2 receptor (PTH2R) constitute a new endocrine system with conserved neuronal mRNA expression in zebrafish and mouse brain. The functions associated with the murine TIP39-PTH2R system include nociception and regulation of hypothalamic (GnRH) and pituitary hormones (ACTH, GH, and LH). Zebrafish studies have shown that TIP39 and the PTH2R are expressed early in brain development in a spatial and temporal relationship with sonic hedgehog (Shh). Furthermore, embryos which have TIR39 expression inhibited by morpholino are not viable and show severely impaired brain development. Thus, the zebrafish TIP39-PTH2R studies suggest that this system serves a crucial role in neuroendocrine development and perhaps other organs such as pancreas and heart where TIR39 and the PTH2R are expressed. Therefore, the zebrafish, Danio rerio, provides a powerful model system to study TIR39-PTH2R-regulated mechanisms during development of the central nervous system (CNS). Hence, molecular analyses are proposed to better elucidate the effects of altered zebrafish PTH2R and TIR39 transcript expression during early brain development. Three aims will be pursued, 1) Evaluation of PTH2R and TIP39 MO-injected embryos during early brain development by examining the expression of Shh, GH, ACTH, LH, and GnRH. 2) Determine the relationship between TIP39 and Shh during brain development by examining the locale of cells expressing TIP39 in embryos which have a deficiency for Shh or Shh signaling (sonic you and slow-muscle-omitted). 3) Evaluation of PTH2R and TIP39 overexpression in the brain and hypothalamus (respectively) during early brain development by examining the expression of the marker genes described above. Because the PTH2R has not been analyzed by gene knockout in any organism, these experiments will provide novel insights into the role of this new TIP39-PTH2R system in normal and perturbed embryos. This work is part of a long-term molecular developmental-genetic investigation on the mechanistic role(s) of TIP39-PTH2R system during CNS, and other organ, development.