|Budget Amount *help
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1996: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1995: ¥1,500,000 (Direct Cost: ¥1,500,000)
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
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