永岡 明伸 武田薬品工業株式会社, 創薬第一研究所, 所長
FURUKAWA Yoshiko Gifu Pharmaceutical University Pharmacy, Associate, 薬学部, 助教授 (20219108)
FURUKAWA Shoei Gifu Pharmaceutical University Pharmacy, Associate Professor, 薬学部, 助教授 (90159129)
KOHNO Michiaki Gifu Pharmaceutical University Pharmacy, Associate Professor, 薬学部, 助手 (00027335)
NAGAOKA Akinobu Takeda Pharmaceutical Industries, Ltd.Discovery Research Laboratories I,Head
|Budget Amount *help
¥11,500,000 (Direct Cost : ¥11,500,000)
Fiscal Year 1994 : ¥5,000,000 (Direct Cost : ¥5,000,000)
Fiscal Year 1993 : ¥6,500,000 (Direct Cost : ¥6,500,000)
Alzheimer's disease (AD) leads to a progressive and irreversible damage of memory and cognitive function in afflicted elderly individuals. AD is characterized by various pathological markers in the brain, chief of which are large amounts of amyloid plaques and neurofibrillary tangles and neuronal cell loss. Especially, cholinergic neurons of basal forebrain nuclei, which are important for cognitive functions, undergo degenerative changes in AD.Besides, in AD,the activety of acetylcholine esterase and choline acetyltransferase are reduece and, as a results, the amount of acetylcholine, a neurotransmitter, is also reduced. At present, litte is known about the relationship between AD and neurotrophic factors (NTFs) and other growth factors related to the pathogenesis of AD.The main reason seems to be lack of aboundance of these factors in most tissues, and lack of a method to reliably detect the protein levels of these factos. Purpose of this reseach was thus to develop the highly sentiti
ve, specific enzyme immunoassay (EIA) systems for various NTFs and other cell growth factors related to the pathogenesis of AD and to apply these developed EIA systems for the diagnosis of the AD.Major results are summarized as follows :
(1) we prepared anti-recombinant human NGF antibody IgG and characterized its properties immunologically. Using this antibody IgG,we developed a sentive two-site EIA system for human NGF.We measured NGF levels using this system and found no difference in NGF level in serum, brain-spinal fluid, or brain (hippocampus and parietal cortx) obtained from normal people and patients with AD.These results suggest that a decrease in the NGF level is not causative factor of AD.
(2) We developed an EIA system for NT-3, based on a biotin-streptoavidin detection system capable of measureing concentrations as low as 0.5 pg/ml with high reproducibility. We measured NT-3 levels in the developing rat nervous system as well as in normal subjects and patients with AD.The developmental changes of NT-3 level in rat brain and peripheral tissues coincided with the changes in NT-3 mRNA level reported before. We could not find any differences in NT-3 level in serum, cerebrospinal fluid, or brain (hippocampus and parietal cortex) obtained from controls and patients with AD.