Purification and Characterization of Neurotrophic Factor for Retinal Cholinergic Neurons Derived from Cultured Hippocampal Neurons

The Japanese Journal of Pharmacology

Kyoko Omori¹  Masafumi Inoue¹  Chiyoko Inagaki¹  Masanobu Uyama²  Tohru Yasukura¹  Toshiko Mikami¹ 

[1] Departments of Pharmacology, Kansai Medical University;Departments of Ophthalmology, Kansai Medical University

关键词: Hippocampus;    Neurotrophic factor;    Choline acetyltransferase;    Amacrine cell (retina);    Peptide;   

DOI  :  10.1254/jjp.65.153

学科分类：药理学

来源: Nihon Yakuri Gakkai Henshuubu / Japanese Pharmacological Society

【 摘 要 】

References(32)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.

【 授权许可】

Unknown