DEVELOPMENT OF NEURONAL CELL SURFACE GLYCOPROTEINS

神经细胞表面糖蛋白的发育

基本信息

批准号：
3397184
负责人：
MICHAEL L SHELANSKI
金额：
$ 20.14万
依托单位：
COLUMBIA UNIV NEW YORK MORNINGSIDE
依托单位国家：
美国
项目类别：
财政年份：
1987
资助国家：
美国
起止时间：
1987-09-01 至 1989-08-31
项目状态：
已结题

项目摘要

Our aim is to define the role of cell surface glycoproteins (GP) in nervous system development and function. Work will continue on the purification and characterization of the principal antigenic cell surface and released GP of PC12 cells and sympathetic neurons, two homogeneous, well characterized model neuronal systems. A number of antigenic cells surface glycoproteins also present in brain have been identified on PC12 cells and sympathetic neurons including those at apparent Mr = 230, 180, 160, 140, 118 and 25 kd. We have studied the 230 kd component extensively and named it after its properties, the NGF-inducible large external (NILE) GP. Specific antibodies (Ab) have been prepared against the NILE and the 180 kd GP and monoclonal and monospecific polyclonal Ab will be produced against the others. The Ab will be coupled to insoluble matrices and for the immunoaffinity purification of each of these macromolecules. The Ab will also be used to identify cross-reactive proteins in brain which will be purified and compared to their membrane bound and released PC12 and sympathetic neuron counterparts. The polyclonal sera will be used in conjunction with trypsin-treatment of the intact cells and tunicamycin-inhibition of glycosylation to determine the structure of each of these components with respect to the cell membrane. The Ab will also be used in anatomical localization studies in cultured and in fixed CNS, PNS and adrenals of different developmental ages. in an attempt understand the role of these GP, the effects of the antisera and the purified GP will be assessed on a variety of neuronal behaviors. Immunological methods will be used to select mutants of PC12 cells which lack or are altered in specific cell surface GP and these will also be screened for functional changes. Transplacental exposure of embryos to Ab against the individual GP will be studied to understand the developmental role of these molecules. Finally a radioimmunoassay for the released GP will be developed for immunodiagnosis of neural crest tumors in animals.

我们的目标是确定细胞表面糖蛋白 (GP) 在神经系统中的作用系统的开发和功能。净化工作将继续进行主要抗原细胞表面和释放的表征 PC12细胞和交感神经元的GP，两个均质，良好表征模型神经元系统。表面有许多抗原细胞 PC12 细胞上也发现了大脑中也存在的糖蛋白交感神经元，包括表观 Mr = 230、180、160、140 处的交感神经元， 118 和 25 公里。我们广泛研究了 230 kd 组件并将其命名为根据其特性，NGF 诱导型大外部 (NILE) GP。已制备针对 NILE 和 180 kd 的特异性抗体 (Ab) 将生产 GP 以及单克隆和单特异性多克隆抗体其他人。 Ab 将与不溶性基质偶联，并用于对这些大分子中的每一个进行免疫亲和纯化。抗体将也可用于识别大脑中的交叉反应蛋白纯化并与膜结合和释放的 PC12 和交感神经元对应物。多克隆血清将用于与完整细胞的胰蛋白酶处理相结合，衣霉素抑制糖基化以确定每个结构这些成分相对于细胞膜的影响。 Ab 也将是用于培养和固定 CNS、PNS 的解剖定位研究以及不同发育年龄的肾上腺。试图理解这些 GP 的作用、抗血清和纯化 GP 的作用将是评估各种神经元行为。免疫学方法将用于选择 PC12 细胞的突变体，这些突变体缺乏或改变了特定的细胞表面GP等也将进行功能变化筛查。胚胎经胎盘暴露于针对个体 GP 的抗体将研究以了解这些分子的发育作用。最后一个已发布的GP的放射免疫分析将被开发用于免疫诊断动物神经嵴肿瘤。

项目成果

期刊论文数量（9）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Nerve growth factor-inducible large external (NILE) glycoprotein: studies of a central and peripheral neuronal marker.

神经生长因子诱导性大外部 (NILE) 糖蛋白：中枢和外周神经元标记物的研究。

DOI：
10.1523/jneurosci.03-03-00441.1983
发表时间：
1983
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
0
作者：
Salton,SR;Richter-Landsberg,C;Greene,LA;Shelanski,ML
通讯作者：
Shelanski,ML

Isolation of NILE glycoprotein-related cDNA probes.

NILE 糖蛋白相关 cDNA 探针的分离。

DOI：
10.1111/j.1471-4159.1987.tb00958.x
发表时间：
1987
期刊：
Journal of neurochemistry
影响因子：
4.7
作者：
Sajovic,P;Ennulat,DJ;Shelanski,ML;Greene,LA
通讯作者：
Greene,LA

Biochemical properties of the nerve growth factor-inducible large external (NILE) glycoprotein.

神经生长因子诱导性大外部 (NILE) 糖蛋白的生化特性。

DOI：
10.1523/jneurosci.03-12-02420.1983
发表时间：
1983
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
0
作者：
Salton,SR;Shelanski,ML;Greene,LA
通讯作者：
Greene,LA

Regional brain glucose use in unstressed rats after two days of starvation.

饥饿两天后无应激大鼠的局部脑葡萄糖使用情况。