ONTOGENY OF FOREBRAIN DOPAMINE RECEPTORS

PI将在该奖项的支持下学习以下内容： 使用D2、D3和D4多巴胺受体亚型探针的杂交 结合脑提取mRNA的北方印迹分析; 免疫组织学技术来鉴定对应的细胞群 配体结合或探针杂交的领域;以及，详细的 发育中的神经系统的大体和显微神经解剖学知识 人脑这些获得的技能将用于补充 表征[3H]-YM个体发育的正在进行的工作的发现 09151 - 2在人胎脑组织中的结合，使用受体 放射自显影PI新获得的技能将应用于 以下具体项目使用的组织从中央 华盛顿大学人类胚胎学实验室：1） 人前脑多巴胺受体的个体发育特征 使用定量受体放射自显影术来识别大体和微观- 特异性结合的神经解剖学分布; 2）比较 细胞表面多巴胺受体的分布 D2、D3和D4多巴胺受体信息原位杂交; 3） 表达多巴胺受体的细胞群的鉴定 （表型和/或信息）在早期至中期的前脑中 妊娠期使用适当的组织学技术; 4）澄清 亚板神经元（NGFR）之间的关系 免疫组织化学）对表达多巴胺的细胞群 受体;和5）在试点项目中识别特定组织 被认为存在先天性脑异常风险的样本，以与 没有这样的风险。监督研究经验提供了 通过这个奖项将使PI能够描述"正常"个体发育 人类前脑多巴胺受体。

The PI will learn the following with the support of this award: in situ hybridization using probes for D2, D3 and D4 dopamine receptor subtypes in conjunction with Northern blot analysis of brain-extracted mRNA; immunohistological techniques to identify cell populations corresponding with the areas of ligand binding or probe hybridization; and, a detailed knowledge of the gross and microscopic neuroanatomy of the developing human brain. These acquired skills will be used to complement the findings of work-in-progress characterizing the ontogeny of [3H]-YM 09151-2 binding in human fetal brain tissue using receptor autoradiography. The PI's newly acquired skills will be applied to the following specific projects using tissues obtained from the Central Laboratory for Human Embryology at the University of Washington: 1) characterization of the ontogeny of dopamine receptors in human forebrain using quantitative receptor autoradiography to identify gross and micro- neuroanatomic distributions of specific binding; 2) comparison of the distribution of cell-surface dopamine receptors to the distribution of D2, D3 and D4 dopamine receptor message using in situ hybridization; 3) identification of the cell populations which express dopamine receptors (phenotype and/or message) int he forebrain during early-to-middle gestation using appropriate histological techniques; 4) clarification of the relationship of subplate neurons (identified by NGFR immunohistochemistry) to cell populations which express dopamine receptors; and 5) identification, in a pilot project, specific tissue specimens considered at risk for congenital brain anomaly to compare to tissues without such risk. The supervised research experience provided by this award will enable the PI to characterize the "normal" ontogeny of human forebrain dopamine receptors.