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TROPHIC MATERIAL IN NEURONAL TISSUE

神经组织中的营养物质

基本信息

批准号：
3845302
负责人：
J ZHU
金额：
--
依托单位：
NATIONAL INSTITUTE OF MENTAL HEALTH
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/3845302
关键词：
PC12 cells adrenal glands adrenal medulla axon chromaffin cells dendrites growth media laboratory rat macroglobulins molecular genetics neurochemistry neurotrophic factors protease inhibitor protein purification tissue /cell culture trypsin inhibitors

项目摘要

During the development of a primary culture of chromaffin cells from adult rat adrenals, neurite outgrowth from chromaffin cells was sometimes observed. Subsequently we collected the adrenal medullary culture media in which chromaffin cell neurite outgrowth was observed and found that it stimulated neurite out-growth from cultured adrenal chromaffin cells and also from PC-12 cells. The experimental results to date indicate that this neurite out-growth activity is a heat labile protein but is distinct from NGF, aFGF, bFGF, EGF and IGF-I. Recently, it has been reported that various protease inhibitors are capable of initiating neurite-outgrowth from various cells. Because of this, various protease inhibitors, alpha2-macroglobulin, soybean trypsin inhibitor, alpha1-antitrypsin, aprotinin, leupeptin and antipain were examined and found to exert no neurite-outgrowth activity on PC- 12 cells. Our previous experiments also indicate that adrenal medullary non-chromaffin cells may release this neurite-outgrowth activity into the culture media during culture. In this study, large quantities of adrenal medullary cells were prepared for extractions of the neurite outgrowth promoting factor(s) and some extracts were found to be active in initiating neurite-outgrowth from PC-12 cells. We plan to establish an optimal extraction condition and then to isolate this neurite-outgrowth factor(s) for further characterization.

在嗜铬细胞的原代培养过程中，在成年大鼠肾上腺中，有时嗜铬细胞会长出神经突观察随后，我们收集肾上腺髓质培养基，其中嗜铬细胞神经突生长被观察到，并发现它刺激培养的肾上腺嗜铬细胞的神经突生长，也来自PC-12细胞。迄今为止的实验结果表明，这种神经突生长活性是一种热不稳定蛋白质， EGF、bFGF、EGF、IGF-1。近日，有报道称，各种蛋白酶抑制剂能够启动神经突生长从不同的细胞。因此，各种蛋白酶抑制剂， α 2-巨球蛋白，大豆胰蛋白酶抑制剂，α 1-抗胰蛋白酶，抑肽酶、亮抑酶肽和抗痛剂进行了检查，对PC- 12细胞的神经突生长活性。我们之前的实验也表明肾上腺髓质非嗜铬细胞可以释放在培养过程中将这种神经突生长活性转移到培养基中。本研究制备了大量的肾上腺髓质细胞，用于提取神经突生长促进因子和一些提取物被发现在启动神经突生长中具有活性， PC-12细胞我们计划建立最佳提取条件，然后分离这种神经突生长因子，特征化