U.S.-Argentina Cooperative Research: Conditional Gene Targeting of Brain Dopamine Receptors

9901278LowThis America Program奖将资助俄勒冈州波特兰市俄勒冈健康科学大学伏勒姆研究所的马尔科姆·洛博士和阿根廷布宜诺斯艾利斯大学的马塞洛·鲁宾斯坦博士之间为期三年的合作研究项目。他们计划利用一项新技术，在成年小鼠的选择性脑底物中产生有条件的多巴胺受体亚型的敲除。哺乳动物大脑中这些突变的目的是提供对生理过程的实验控制。在胚胎干细胞中通过同源重组产生零等位基因是确定基因在体内的生理功能的一项强有力的技术。然而，这种方法的缺点包括单个基因的多种功能的丧失和特定神经元中其他基因表达过程的变化对发育的补偿。在这个项目中，研究人员将开发一种新技术来克服这些限制。如果在多巴胺能系统中取得成功，同样的实验策略将普遍适用于大脑中表达的许多基因的功能分析。两名研究人员是将要使用的技术方面的专家，因此这项工作将产生重大的新突变方法、小鼠品系和对多巴胺受体功能的见解。这项研究的更广泛的影响包括，除了开发新的方法来产生中枢神经系统基因的局部条件突变外，还包括了解五种不同的多巴胺受体亚型在局部脑区的功能意义*

9901278LowThis Americas Program award will fund a three year cooperative research project between Dr. Malcolm Low, Vollum Institute, Oregon Health Sciences University, Portland, Oregon, and Dr. Marcello Rubinstein, Universidad de Buenos Aires, Argentina. They plan to utilize a new technology that will produce a conditional knockout of dopamine receptor subtypes in selective brain substrates of the adult mouse. The objective of these mutations in mammalian brain is to provide the experimental control of physiological processes. The generation of null alleles by homologous recombination in embryonic stem cells is a powerful technique to determine the physiological function of genes in vivo. However, drawbacks of this approach include the loss of multiple functions of a single gene and developmental compensation by changes in the expression of other genes processes in specific neurons. In this project the investigators will develop a new technology to overcome these limitations. If successful in the dopaminergic system, the same experimental strategy would be generally applicable to the functional analysis of many genes expressed in the brain.The two investigators are experts in the techniques to be employed, therefore significant new mutational approaches, mouse strains and insights into dopamine receptor function will result from this work. The broader impact of this research includes, besides the development of new methods to produce localized, conditional mutations in central nervous systems genes, the understanding the functional significance of the five different dopamine receptor subtypes in localized brain regions ***