HIV AND PEPTIDERGIC NEURONS IN BRAIN CULTURES

本申请的目标是阐明细胞和 HIV包膜蛋白的细胞内分子机制 gp 120诱导脑内肽能神经元的功能障碍和/或丧失， 大脑，重点是最丰富的肽能神经元， 神经肽Y（NPY）神经元。 两个工作假设将受到挑战。 第一，gp 120诱导神经细胞信号转导缺陷和神经元死亡。 艾滋病患者的大脑，这是由于，部分， gp 120在神经元本身和/或其他细胞上的分泌产物 调节神经元功能 第二，gp 120诱导了一种慢性的 Fos/Jun抑制性转录因子复合物的表达 在NPY神经元和/或调节NPY神经元的其它细胞内， 其结果是有缺陷的信号转导，例如，受损 对cAMP/蛋白激酶C通路的激活的反应。 一个 来自胎鼠的聚集培养物的实验模型系统 大脑在我们的实验室里完全运作，将被用作一种工具， 实现我们的目标。 为了验证大鼠模型可以作为 作为人类的替代品，我们将建立一个文化体系， 人类流产的大脑，并进行了一项平行的研究gp 120 在人类和大鼠模型中的作用。 S.A.1. 建立文化体系， 一个指导方针。 S.A.2.描述gp 120对 培养的NPY神经元的功能/存活：进行比较 在人类和大鼠模型系统中进行研究。 a）定义时间/剂量 GP 120诱导功能障碍的动力学（对毛喉素和 佛波酯）和NPY神经元死亡。B）定义结构-功能 gp 120作用的特点：gp 41的作用， VIP/CD 4样受体与gp 120特定区域的需求。 c）确定胶质细胞是否介导gp 120对NPY的作用 神经元 S.A. 3.阐明细胞内的分子机制 gp 120诱导的信号转导缺陷，重点是Fos/Jun 转录调节因子家族。 这些研究产生的结果将提供深入了解 gp 120在人脑中的作用机制，这将有助于 作为未来治疗剂设计的基线/指南 临床管理和可能的预防异常表达 在HIV感染者的大脑中。 一个重要而及时的 本研究的结果将是一种可靠的体外静脉给药系统的可用性。 模型系统来测试这些新设计的治疗效果 剂.

The goals of this application are to elucidate the cellular and intracellular molecular mechanisms by which the HIV envelope protein gp120 induces dysfunction and/or loss of peptidergic neurons in the brain, with emphasis on the most abundant peptidergic neuron, the neuropeptide Y (NPY) neuron. Two working hypotheses will be challenged. One, gp120 induces defective signal transduction and neuronal death in the brain of AIDS patients and this is due, in part, to an effect of gp120 on the neuron itself and/or other cells the secretory products of which regulate neuronal function. Two, gp120 induces a chronic expression of an inhibitory transcription factor complex of Fos/Jun within the NPY neuron and/or other cells regulating the NPY neuron, a consequence of which is defective signal transduction, e.g., impaired response to activation of the cAMP/protein kinase C pathways. An experimental model system of aggregate cultures derived from fetal rat brain is fully operative in our laboratory and will be used as a tool to achieve our goal. To validate that the rat model can serve as an alternative to the human, we will establish a culture system derived from the brains of human abortuses and conduct a parallel study of gp120 action in the human and rat models. S.A.1. Establish culture system as a guideline. S.A.2. Characterize the effects of the gp120 on the function/survival of the cultured NPY neurons: conduct a comparative study in the human and rat model systems. a) Define the time/dose kinetics of gp120 induction of dysfunction (response to forskolin and phorbol ester) and death of NPY neurons. b) Define the structure-function characteristics of gp120 actions: the role of gp41, the role of a VIP/CD4-like receptor and the requirement for specific regions of gp120. c) Ascertain if glial cells mediate the effects of gp120 on the NPY neurons. S.A. 3. Elucidate the intracellular molecular mechanisms of gp120-induced defective signal transduction, with emphasis on Fos/Jun family of transcription regulators. The results generated in these studies will provide insight into the mechanisms underlying gp120 actions in the human brain, which will serve as the baseline/guide for the design of therapeutic agents for future clinical management and possibly prevention of abnormalities expressed in the brains of HIV-infected people. An important and most timely outcome of this study will be the availability of an iv vitro reliable model system to test the efficacy of these newly designed therapeutic agents.