FETAL TISSUE TRANSPLANT AND OTHER GENE EXPRESSION SYSTEMS

该组件中描述的实验的总体目标是 描述胎儿组织的生长、发育和功能 从有精神分裂症风险的流产中分离出的碎片，以及 确定这些组织之间是否存在功能差异 和正常对照组织，可能会露出一些生物花瓶 治疗精神分裂症。这些实验有四个基本目标。这个 第一个目标将是研究移植的脑区(例如 海马区和大脑皮层)取自正常和 将精神分裂症患者转化为裸鼠，并探讨其作用机制 在我们的增长和成熟率下降的基础上 观察到的。这些实验将测试非遗传性(例如，药物- 相关)以及这些差异背后的遗传机制。在……里面 特别是，我们将测试神经营养的特殊异常 如NT-3，它主要定位于海马体，或者 GDNF，对多巴胺神经元有特殊的营养作用。第二 主要目标将是记录脑电生理活动 海马区、大脑皮层等异种移植中的脑细胞 正常女性和精神分裂症患者胎儿组织的脑区 眼科和体外培养。细胞内和细胞外记录将被 采用以及全细胞斑片技术来表征固有的 这些神经元的特性以及它们对神经递质的反应。这个 移植也将通过免疫组织化学和原位研究。 杂交以评估特定类型神经元的发育。 第三个主要目标将是在眼科和体外建立 假设的电路(基于正在进行的其他 该中心的组成部分)对感官缺陷负责 精神分裂症患者中出现的门控。人胎脑序贯异种移植 将结合眼球中的区域，试图定义哪个神经元 元素必须来自流产的脑组织碎片 精神分裂症患者在发育或功能上产生缺陷 这些电路。最终目标将是研究分离的细胞， 已经被从正常和成人中提取的遗传物质中 精神分裂症尸检脑组织的可能特征 神经递质受体的差异。候选基因将是 在非洲爪哇卵母细胞中表达，在人胚胎肾细胞系(HEK)中表达 293细胞)、雪旺细胞和原代培养的成纤维细胞。 正常人、精神分裂症患者和专职携带者。在所有这些牢房中 系统，基因的表达将通过细胞内和补丁来研究 记录技术和特定受体亚基的表达 将在生化上得到证实。

The overall objectives of the experiments described in the component are to characterize the growth, development, and function of fetal tissue fragments isolated from abortuses at risk for schizophrenia, and to determine whether there are functional differences between such tissue and normal control tissue that might reveal some of the biological vases for schizophrenia. There are four basic aims of these experiments. The first objective will be to study transplanted brain regions (e.g. hippocampus and cerebral cortex) obtained from fetal tissue of normal and schizophrenic women into athymic rats, and to examine the mechanisms underlying the reduced growth and rate of maturation that we have observed. These experiments will test for non-genetic (e.g., medication- related) as well as genetic mechanisms underlying these differences. In particular, we will test for specific abnormalities in neurotrophic factors such as NT-3, which is primarily localized to hippocampus, or GDNF, which has specific trophic effects on dopamine neurons. The second major objective will be to record the electrophysiological activity of brain cells in xenotransplants of hippocampus, cerebral cortex, and other brain areas from fetal tissue of normal and schizophrenic women both in oculo and in vitro. Intracellular and extracellular recording will be employed as well as whole cell patch techniques to characterize intrinsic properties of these neurons and their response to neurotransmitters. The transplants will also be studied by immunohistochemistry and by in situ hybridization to assess the development of specific types of neurons. The third major objective will be to establish in oculo and in vitro the circuits hypothesized (on the basis of ongoing studies in other components of the Center) to be responsible for the deficit in sensory gating seen in schizophrenia. Sequential xenografts of human fetal brain areas in oculo will be combined in an attempt to define which neuronal elements must be derived from brain tissue fragments from abortuses of schizophrenics to produce deficits in the development or function of these circuits. The final objective will be to study isolated cells that have been transfected with genetic material derived from normal and adult schizophrenic post mortem brain tissue to characterize possible differences in neurotransmitter receptors. Candidate genes will be expressed in Xenopus oocytes, in a human embryonic kidney cell line (HEK 293 cells), in Schwann cells, and in primary cultures of fibroblasts from normals, schizophrenics and obligate carriers. In all these cell systems, gene expression will be studied by intracellular and patch recording techniques, and the expression of specific receptor subunits will be confirmed biochemically.