NONCLASSICAL EXPORT CHICKEN CILIARY NEUROTROPHIC FACTOR

描述：（来自申请人摘要）多细胞生物体具有 协调其不同细胞活动的复杂机制， 器官，需要某些类型的细胞释放信号分子， 结合这些分子并对这些分子作出适当反应能力 类型鸡睫状神经营养因子（chCNTF）是一种蛋白质， 鸟类神经系统的正常发育。由神经支配的细胞产生 睫状神经节神经元，chCNTF必须由它们释放以结合， 并调节神经元的发育。 CNTF缺乏一个N-末端序列，该序列将蛋白质靶向到一个 细胞中的特征性分泌途径;先前的研究表明， 不是通过这种“经典”途径分泌的。这项提案的目的是 从而表征其分泌方式。这些成果将意义重大 在阐明真核细胞用于通信的基本方法中， 因为CNTF和相关蛋白质对于正常的胚胎发育是必需的， 发展，可以确定发展缺陷和/或潜在的来源 增强细胞间通讯的操作。 CNTF输出中涉及的细胞区室将通过以下方式进行鉴定：1） CNTF在培养细胞中的代谢标记，随后进行细胞分级 和其所在的隔室的表征; 2）附着 将绿色荧光蛋白转化为CNTF， 细胞;和3）分离细胞器并评估它们摄取CNTF的能力 体外将其靶向至输出途径所必需的chCNTF区域将 通过检测chCNTF与大鼠的各种杂交能力， CNTF（一种不分泌的密切相关的蛋白质）由细胞输出。

DESCRIPTION: (from applicant's abstract) Multicellular organisms possess complex mechanisms that coordinate the activity of their diverse cells and organs, entailing the release of signaling molecules by some types of cells and the ability to bind and respond appropriately to these molecules by other types. Chicken ciliary neurotrophic factor (chCNTF) is a protein required for proper development of the avian nervous system. Produced by cells innervated by ciliary ganglion neurons, chCNTF must be released by them in order to bind to, and mediate development of, the neurons. CNTF lacks an N-terminal sequence that targets proteins into a well-characterized secretory pathway in cells; prior studies indicate that it is not secreted through this "classical" pathway. The goal of this proposal is thus to characterize its means of secretion. These results will be significant in clarifying the basic methods used for communication by eukaryotic cells and, because CNTF and related proteins are essential for proper embryonic development, may identify sources of developmental defects and/or potential manipulations that enhance intercellular communication. The cellular compartments involved in CNTF export will be identified by: 1) metabolic labeling of CNTF in cultured cells followed by fractionation of cells and characterization of the compartments in which it resides; 2) attaching green fluorescent protein to CNTF to allow it to be followed through living cells; and 3) isolating organelles and assessing their ability to take up CNTF in vitro. The region of chCNTF necessary to target it to an export pathway will also be identified by testing the ability of various hybrids of chCNTF and rat CNTF (a closely related protein that is not secreted) to be exported by cells.