SVP38: A Synaptic Vesicle Protein Whose Appearance Correlates Closely with Synaptogenesis in the Rat Nervous System
SVP38: A Synaptic Vesicle Protein Whose Appearance Correlates Closely with Synaptogenesis in the Rat Nervous System
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SVP38：一种突触小泡蛋白，其外观与大鼠神经系统突触发生密切相关
DOI：
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发表时间：
1987
期刊：
影响因子：
--
通讯作者：
C. Barnstable
中科院分区：
文献类型：
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作者：
S. Devoto;C. Barnstable
与突触小泡特异性相关的分子显然是执行突触小泡某些特殊功能的候选分子。突触小泡的几种蛋白质已被鉴定，但没有一个具有经过证实的功能。突触蛋白 I 被确定为突触小泡的主要磷蛋白，可以调节小泡释放的可用性。在单克隆抗体的帮助下已经鉴定了其他小泡蛋白，包括 65,000 Da 跨膜蛋白 (p65)：和 100,000 Da 跨膜糖蛋白。”我们和其他人已经表征了一种 38,000 Da 的突触小泡蛋白，该蛋白被命名为 p38、“突触素”和 SVP38.6。我们针对这种蛋白生产的单克隆抗体对整个大鼠神经系统的突触结构进行了染色。基于已建立的程序(图1A)，对均质化的大鼠大脑皮层进行亚细胞分级分离，以鉴定SVP38在细胞内的位置。该分级分离的结果显示抗原在已知含有突触小泡的级分中一致富集(图1B)。通过SVP38和突触蛋白I的免疫印迹分析来自受控孔玻璃色谱的级分，这是分离突触小泡的最后一步。两种蛋白质显示相同的洗脱曲线(图1C)，证明SVP38也是突触小泡蛋白质。了解突触小泡蛋白功能的一种方法是将其发育外观与突触前功能的其他特征的外观进行比较。仅在突触形成后表达的分子预计将执行与突触形成前也在生长锥中表达的分子不同的功能。我们使用点免疫结合试验来分析三种突触小泡蛋白在大脑皮层中的发育表达：突触蛋白 I、p65 和 SVP38。这些数据以及使用电子显微镜测量的有关皮层突触形成的可比信息如图 2 所示。8 SVP38 水平随着突触形成而增加，而突触蛋白 I 和 p65 较早表达，并且在突触发生期间不会增加到相同程度。由于突触形成的电子显微镜分析依赖于电子致密突触后密度的出现，因此突触形成的初始阶段可能先于 SVP38 的表达。突触蛋白 I、p65 和 SVP38 之间发育表达的差异
Molecules specifically associated with synaptic vesicles are obvious candidates to carry out some of the specialized functions of synaptic vesicles. Several proteins of the synaptic vesicle have been identified, however, none of them have a proven function. Synapsin I was identified as a major phosphoprotein of the synaptic vesicle and may regulate the availability of vesicles for release.’ Other vesicle proteins have been identified with the aid of monoclonal antibodies, including a 65,000-Da transmembrane protein (p65): and a 100,000-Da transmembrane glycoprotein.” We and others have characterized a 38,000-Da synaptic vesicle protein that has been named p38,’ synaptophysin,’ and SVP38.6 The monoclonal antibody we produced against this protein stained synaptic structures throughout the rat nervous system. Subcellular fractionation of homogenized rat cerebral cortex, based on established procedures’ (FIG. lA), was undertaken to identify the location of SVP38 within the cell. The results of this fractionation showed the antigen to be consistently enriched in the fractions known to contain synaptic vesicles (FIG. 1B). The fractions from the controlled pore glass chromatography, the last step in the isolation of synaptic vesicles, were analyzed by immunoblotting for SVP38 and synapsin I. Both proteins showed the same elution profile (FIG. lC), demonstrating that SVP38 is also a synaptic vesicle protein. One approach to understanding the function of synaptic vesicle proteins is to compare their developmental appearance with the appearance of other features of presynaptic function. Molecules expressed only subsequent to synapse formation can be expected to carry out different functions from those also expressed in the growth cone prior to synapse formation. We have used a dot immunobinding assay to analyze the developmental expression in the cerebral cortex of three synaptic vesicle proteins: synapsin I, p65, and SVP38. These data are shown in FIGURE 2, along with comparable information on synapse formation in the cortex, as measured using the electron microscope.8 SVP38 levels increase in parallel with the formation of synapses, while synapsin I and p65 are expressed earlier and do not increase to the same extent during synaptogenesis. Since the electron microscope analysis of synapse formation relied on the appearance of the electron-dense postsynaptic density, it is possible that the initial stages of synapse formation precede the expression of SVP38. The difference in developmental expression between synapsin I, p65, and SVP38
DOI：
10.1073/pnas.82.12.4137
发表时间：
1985-01-01
影响因子：
11.1
作者：
JAHN, R;SCHIEBLER, W;GREENGARD, P
通讯作者：
GREENGARD, P
DOI：
10.1073/pnas.82.9.3035
发表时间：
1985-01-01
影响因子：
11.1
作者：
LLINAS, R;MCGUINNESS, TL;GREENGARD, P
通讯作者：
GREENGARD, P