Increased levels of GAP-43 protein in schizophrenic brain tissues demonstrated by a novel immunodetection method.
Increased levels of GAP-43 protein in schizophrenic brain tissues demonstrated by a novel immunodetection method.
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一种新型免疫检测方法证明精神分裂症脑组织中 GAP-43 蛋白水平升高。
DOI：
10.1007/bf03160108
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
Perrone-Bizzozero,NI
中科院分区：
文献类型：
--
作者：
Sower,AC;Bird,ED;Perrone-Bizzozero,NI
神经和精神疾病的分子基础研究往往依赖于脑组织中特定蛋白质的精确测定。在这项研究中，我们开发了一种方法来测量人类死后大脑标本中神经特异性生长相关蛋白GAP-43的水平。这种快速定量的方法是基于免疫检测程序。简单地说，通过点印迹装置将突触体质膜（SPMs）沉积到聚偏二氟乙烯（PVDF）膜上，然后使用单特异性多克隆抗体进行特异性GAP-43检测。总的来说，与Western blot和一维和二维聚丙烯酰胺凝胶相比，点印迹方法有几个优点。该方法灵敏度高，重复性好，可同时快速测定多个样品。使用这种技术，我们检测了精神分裂症患者和年龄、性别和死后间隔（PMI）匹配的对照组的Brodmann区17、20和10的GAP-43蛋白水平。这些研究表明，精神分裂症患者大脑视觉关联和额叶皮层（20区和10区）的GAP-43水平有所增加。考虑到GAP-43的表达与神经连接的建立和重塑之间的关系，我们的研究结果支持了精神分裂症与人类大脑联合区突触连接组织紊乱有关的假设。
Studies on the molecular basis of neurological and psychiatric disorders often rely on the precise determination of specific proteins in brain tissues. In this study, we have developed a method for measuring the levels of the neural-specific growth-associated protein, GAP-43, in human postmortem brain specimens. This rapid and quantitative method is based on immunodetection procedures. Briefly, synaptosomal plasma membranes (SPMs) are deposited onto polyvinylidene difluoride (PVDF) membranes via a dot-blotting apparatus, followed by specific GAP-43 detection using a monospecific polyclonal antibody. Overall, the dot-blot procedure provided several advantages over Western blots and one-dimensional and two-dimensional polyacrylamide gels. The assays were more sensitive, reproducible, and allowed the rapid and simultaneous determination of multiple samples. Using this technique, we examined the levels of the GAP-43 protein in Brodmann’s areas 17, 20, and 10 of schizophrenic and age-, sex- and postmortem interval (PMI) matched controls. These studies revealed an increase in the levels of GAP-43 in visual association and frontal cortices (areas 20 and 10) of schizophrenic brains. Given the relationship of GAP-43 expression with the establishment and remodeling of neural connections, our results support the hypothesis that schizophrenia is associated with a perturbed organization of synaptic connections in associative areas of the human brain.