HIV AND PEPTIDERGIC NEURONS IN BRAIN CULTURES

这个应用程序的目标是阐明细胞和 HIV包膜蛋白在细胞内的分子机制 Gp120诱导大鼠脑内多肽能神经元功能障碍和/或丢失 大脑，重点是最丰富的肽能神经元， 神经肽Y(NPY)神经元两个可行的假设将受到挑战。 其一，gp120诱导脑内信号转导缺陷和神经元死亡 艾滋病患者的大脑，这在一定程度上是由于 Gp120在神经元本身和/或其他细胞上的分泌产物 调节神经功能的物质。第二，gp120诱导慢性 抑制转录因子Fos/Jun复合体的表达 在NPY神经元和/或其他调节NPY神经元的细胞内， 其后果是信号转导有缺陷，例如受损 对cAMP/蛋白激酶C通路激活的反应。一个 胎鼠聚合培养实验模型体系的建立 Brain在我们的实验室中完全运行，并将用作工具 实现我们的目标。要验证大鼠模型是否可以作为 替代人类，我们将建立一种源于文化的体系 并对gp120基因进行平行研究 在人和大鼠模型中的作用。S.A.1。建立文化体系，如 一条指导方针。S.A.2。描述gp120对 培养的NPY神经元的功能/存活：进行比较 在人和大鼠模型系统中进行研究。A)确定时间/剂量 Gp120诱导功能障碍的动力学(对Forsklin和 佛波酯)和NPY神经元的死亡。B)确定结构-功能 Gp120行动的特点：gp41的作用、 VIP/CD4样受体和对gp120特定区域的要求。 C)确定神经胶质细胞是否介导了gp120对NPY的影响 神经元。阐明其胞内分子机制。 Gp120诱导的信号转导缺陷，重点是Fos/Jun 转录调控因子家族。 这些研究产生的结果将为深入了解 人脑中gp120活动的潜在机制，这将有助于 作为未来治疗药物设计的基线/指南 临床处理和可能的异常预防表达 在艾滋病毒感染者的大脑中。一个重要和最及时的 这项研究的结果将是一种可靠的静脉注射体外试验 模型系统来测试这些新设计的治疗方法的疗效 探员们。

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.