Purification and Characterization of Neurotrophic Factor for Retinal Cholinergic Neurons Derived from Cultured Hippocampal Neurons.
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概要
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A neurotrophic factor that supports the development of cholinergic retinal neurons was purified from media conditioned by a primary culture of embryonic hippocampal neurons. Retinal choline acetyltransferase (ChAT), which is located exclusively in amacrine cells, served as a marker for the development of retinal cholinergic neurons. In a serum-free control culture, retinal neurons from 17-day-old rat fetus displayed little increase in the enzyme activity and a low proportion of neurite-bearing cells (15-20%) within 7 days. The conditioned media, when added to the retinal neuron culture, dose-dependently increased ChAT activity and the number of neurite-bearing cells (40-60%), the maximum ChAT activity being ∼sixfold higher than that in the control. The fraction with these stimulatory activities was purified by Sephadex G-15 column chromatography and two times reverse-phase HPLC. The final fraction showed ∼3, 000-fold higher purification as compared with that in the Sephadex G-15 fraction. Gas-phase protein sequencing analysis of the final fraction yielded a peptide sequence: Tyr-Leu-Leu-Pro-Ala-Gln-Val-Asn-Ile-Asp. A synthetic peptide with this sequence dose-dependently stimulated ChAT activity in the retinal cell culture and dissociated cell culture of the septal nucleus. These findings suggest that the developing hippocampal neurons produce a neurotrophic peptide that stimulates the development of cholinergic neurons.
- 公益社団法人 日本薬理学会の論文
著者
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Uyama Masanobu
Department Of Ophthalmology Kansai Medical University
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YASUKURA Tohru
Department of Pharmacology
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MIKAMI Toshiko
Department of Pharmacology
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OMORI Kyoko
Department of Pharmacology
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Inoue Masafumi
Department Of Cardiology National Fukuyama Hospital
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INAGAKI Chiyoko
Departmctit of Pharmacology, Faculty of Medicine, Kyoto University
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Uyama Masanobu
Departments of Ophthalmology, Kansai Medical University
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Omori Kyoko
Departments of Pharmacology, Kansai Medical University
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