Immunohistochemcial investigation of brain-derived neurotrophic factor in the human brain

本研究成功地制备了一种高灵敏度、高特异性的抗BDNF抗体，该抗体可与人、猴和大鼠的BDNF发生反应，识别3.91pmol的BDNF，并在大鼠脑组织匀浆免疫印迹上显示一条约14 kDa的单带。用此抗体检测了BDNF在成年大鼠脑内的分布，发现BDNF在脑内的分布较以往的研究更为广泛。BDNF在神经元胞体和轴突终末有表达，但未发现胶质细胞。在大脑皮层和海马，第2 ~ 6层可见阳性神经元，其中锥体细胞和多极神经元免疫组化染色阳性。阳性细胞的染色强度在细胞间有一定差异。阳性细胞多呈多极形，分布于第2 ~ 6层。通常阳性的点状染色围绕着 ...更多信息 观察到细胞表面，根据形态学，其似乎是终末扣。皮质第一层有大量阳性轴突终末被标记。阳性神经元分布于基底前脑、纹状体、杏仁核、下丘脑、丘脑内侧核、黑质、颅运动核、脑桥核、前角细胞和丘脑前核。在这些区域，阳性细胞的染色强度是不同的。如在下丘脑的许多核团中含有阳性神经元，室旁核和视上核中的神经元染色最强。我们在肌萎缩侧索硬化症患者的脊髓前角中发现了大量的阳性纤维，而对照组的阳性纤维较少。这些发现证实了BDNF在肌萎缩侧索硬化患者前角轴突中的积聚。少

In the present project, we have succeded to raise a highly sensitive and specific antibody against BDNF,which reacts with the human, monkey and rat BDNF.The antibody recognized 3.91 pmol BDNF and stained a single band of approximately 14kda o nhte immunoblot lane which was sampled from the rat brain homogenates. Using the antibody, we examined the immunohistochemcial distribution of BDNF in the adult rat brain, and found the more widespread distribution of BDNF in the rat brain than the previous dscriptions. BDNF was obsrved in the neuronal somata and axon terminal, but not found glial cells. In the cerebral cortex and hippocampus, the posive neurous were observecddin layr 2 through to layr 6, in which pyramidal and multipolar neurous were simmunostained. The staining intensity of positive cells were somewhat varied among cells. The most intensively positive cells were of multipola shaped cells, which were scatterd fromlayr 2 to layr 6. Usually the positive punctate staining around the … More cell surface were oberved, which appeared to be the terminal button based on the morphology. In the first layr of the ccortex, the numerous positive axon terminals were labeled. The brain region in which we have found positive neurons include the basal forebrain, striatum, amygdala, hypothalamus, medial thalamic nuclei, substantia nigara, cranial motor nuclei, pontine nucleus, olve and the anterior horn cells. In these region, the staining intensity of positive cells were varied. For example, in the hypothalamus where many neclei contained the positive neurons, neurosn in the paraventricular and supraoptic nuclei showed the most intens staining.Using the antibody for the postmortem human brain samples, we observed the similar distributions of BDNF.We examined the autopsy samples of spinal cord of patients with amyotrophic lateral sclerosis. We found numerous positive fibers in the anterior horn of the spinal cords of patients with amyotrophic lateral sclerosis, the staining of whic was sparse in the control specimes. These findings suggset the accumulation fo BDNF in the axons in the anterior horn of patients with amyotrophic lateral sclerosis. Less