NONCLASSICAL EXPORT CHICKEN CILIARY NEUROTROPHIC FACTOR

描述：(摘自申请者摘要)多细胞生物拥有 协调不同细胞活动的复杂机制和 器官，需要某些类型的细胞释放信号分子和 通过其他分子结合并适当地对这些分子作出反应的能力 类型。鸡睫状神经营养因子(ChCNTF)是一种蛋白质，是 鸟类神经系统的正常发育。由神经细胞产生 睫状神经节神经元，chCNTF必须由它们释放才能结合， 并调节神经元的发育。 CNTF缺少一个N-末端序列，该序列可以将蛋白质靶向 细胞中特征明确的分泌途径；先前的研究表明，它 并不是通过这种“经典”途径分泌的。这项提议的目标是 从而确定其分泌方式。这些结果将是意义重大的 为了阐明真核细胞用于通讯的基本方法， 因为CNTF和相关蛋白质对正常胚胎是必不可少的 发展，可以确定发展缺陷和/或潜力的来源 增强细胞间通讯的操作。 CNTF出口中涉及的蜂窝舱将通过以下方式确定：1) CNTF在培养细胞中的代谢标记及细胞分级 以及它所在的隔室的特征；2)附着 将绿色荧光蛋白与CNTF结合，使其能够在生命中被跟随 细胞；3)分离细胞器并评估其摄取CNTF的能力 在试管中。将其靶向输出途径所需chCNTF区域将 也可以通过检测不同的chCNTF和大鼠杂交的能力来鉴定 CNTF(一种密切相关的蛋白质，不能分泌)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.