FETAL TISSUE TRANSPLANT AND OTHER GENE EXPRESSION SYSTEMS

胎儿组织移植和其他基因表达系统

• 批准号: 6243058
• 负责人: BARRY J. HOFFER
• 金额: $ 21.21万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-05-01 至 1998-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6243058
• 关键词: GABA receptor; Xenopus oocyte; cerebral cortex; developmental neurobiology; disease /disorder proneness /risk; electrophysiology; eye transplantation; gene expression; hippocampus; human fetus tissue; human tissue; immunocytochemistry; in situ hybridization; innervation; laboratory rat; neural transmission; neuroanatomy; neuropharmacology; neurotrophic factors; nicotinic receptors; phenotype; postmortem; schizophrenia; voltage /patch clamp; xenotransplantation

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.

本部分所述实验的总体目标是 来表征胎儿组织的生长、发育和功能 从有精神分裂症风险的流产者中分离的片段， 确定这些组织之间是否存在功能差异 和正常对照组织，可能会揭示一些生物花瓶 精神分裂症 这些实验有四个基本目标。 的 第一个目标将是研究移植的大脑区域（例如， 海马和大脑皮层）， 精神分裂症妇女到无胸腺大鼠，并检查机制 这也是我们的生长速度和成熟率降低的原因 观察 这些实验将测试非遗传性（例如，药物治疗- 相关）以及这些差异背后的遗传机制。 在 特别是，我们将测试特定的神经营养异常， 因子如NT-3，其主要定位于海马，或 GDNF对多巴胺神经元具有特异性营养作用。 第二 主要目的是记录以下的电生理活动： 海马、大脑皮层和其他组织的异种移植中的脑细胞 正常和精神分裂症妇女胎儿组织的脑区， oculo和in vitro。 细胞内和细胞外记录将是 采用以及全细胞补丁技术来表征内在的 这些神经元的特性及其对神经递质的反应。 的 还将通过免疫组织化学和原位免疫组织化学研究移植物。 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oculo和体外建立 电路假设（基于其他正在进行的研究 该中心的组成部分）负责感官缺陷 在精神分裂症中出现门控。 人胎脑连续异种移植 将结合oculo中的区域，试图确定哪些神经元 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方面产生缺陷 这些电路。 最终的目标是研究分离的细胞， 已经被来自正常和成年人的遗传物质转染 精神分裂症死后的脑组织 神经递质受体的差异。 候选基因将是 在非洲爪蟾卵母细胞、人胚肾细胞系（HEK）中表达 293细胞）、雪旺细胞和来自人成纤维细胞的原代培养物中， 正常人精神分裂症患者和携带者 在所有这些细胞 系统，基因表达将研究细胞内和补丁 记录技术和特定受体亚单位的表达 将在生化方面得到证实