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.

我们的目的是定义细胞表面糖蛋白（GP）在神经中的作用系统开发和功能。工作将继续进行净化和主抗原细胞表面的表征并释放 PC12细胞和交感神经元的GP，两个同质，很好特征模型神经元系统。许多抗原细胞表面在PC12细胞和同情神经元，包括明显MR = 230、180、160、140， 118和25 KD。我们已经广泛研究了230 kD的组件，并命名在其性质之后，NGF诱导的大型外部（尼罗河）GP。针对尼罗河和180 kD制备了特定抗体（AB） GP和单克隆和单特异性多克隆AB将产生针对其他。 AB将与不溶性矩阵相连这些大分子的免疫亲和力纯化。 AB会还用于识别大脑中的交叉反应蛋白，这将是纯化并将其与膜绑定并释放的PC12进行了比较，并且同情神经元对应物。多克隆血清将用于与完整细胞和完整细胞的胰蛋白酶处理结合糖基化糖基抑制穿牙霉素，以确定每种结构这些成分相对于细胞膜。 AB也将是用于培养和固定CNS中的解剖学定位研究，PNS 和不同发育年龄的肾上腺。尝试了解这些GP的作用，抗血清和纯化的GP的影响将是评估各种神经元行为。免疫方法将是用于选择缺乏或改变特定的PC12细胞突变体细胞表面GP以及这些功能变化也将筛选。胚胎对AB的移植对单个GP的接触将是研究以了解这些分子的发展作用。最后将开发用于免疫诊断的释放GP的放射免疫分子动物中神经rest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。