PURIFICATION OF MESSENGER RNAS ENCODING FOR NEUROTROPIC FACTORS IN RAT BRAIN

已经表明，脑损伤诱导神经营养的出现， 病变部位的活动。 神经营养活性可以在 对交感神经元和副交感神经元的作用。 由于这种和其他神经营养物质的潜在重要性， 神经突促进因子在神经精神病学的大部分工作中 分支，我们决定确定我们是否可以使用分子技术 生物学，以更好地了解它们的功能。 该项目的第一步是开发 用于测试神经营养活性及其产生的测定。 不同的技术用于识别病变 脑特异性mRNA。 信使RNA是从细胞壁制备的。 损伤大鼠脑的伤口腔和对照的等同区域。 采用蔗糖密度梯度离心法进行分离。 每个 在非洲爪蟾细胞中， 在神经元培养物中测试神经营养活性。 的分数 来自受损大脑的负责神经营养活性的mRNA被 用作cDNA合成的模板。 cDNA与mRNA杂交 从对照品中减去两种制剂共有的序列。

It has been shown that brain injury induces the appearance of neurotrophic activity at the lesion site. The neurotrophic activity can be assayed in vitro on sympathetic and parasympathic neurons in culture. Because of the potential importance of this and other neurotrophic and neurite promoting factors for much of the work at the Neuropsychiatry Branch, we decided to determine if we could use techniques from molecular biology to get a better understanding of their function. The first steps at the project have consisted in the development of the assays for testing for neurotrophic activity and their production. Different techniques were used for identification of lesioned brain-specific mRNAs. Messenger RNAs were prepared from the walls of the wound cavity of lesioned rat brains and from equivalent areas of controls. They were fractionnated by sucrose density gradient centrifugation. Each fraction was translated into proteins in Xenopus cocytes and the products tested for neurotrophic activity in the neuron cultures. The fraction of mRNA from lesioned brains responsible for the neurotrophic activity was used as a template for cDNA synthesis. The cDNA was hybridized with mRNA from controls to subtract the sequences common to the two preparations.