TRANSGENIC PRAIRIE VOLES TO DISSECT GENETICS/NEURAL CIRCUITRY OF SOCIAL BONDING

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 该项目的目标是开发转基因技术，在基因和细胞水平上选择性地激活和失活调节配对结合的神经系统。这些资源的开发将主要集中在两个方面：1)催产素受体(OXTR)的遗传失活；2)草原田鼠转基因品系中催产素(OT)释放的细胞调控。对于第一个目标，我们将利用Cre-lox和siRNA技术，这些技术已经在突变小鼠中成功使用，以时间调节和细胞类型特定的方式调节基因表达。在过去的一年里，我们已经筛选了几个OXTRshRNA，以确定两个在细胞培养和脑中产生最大击倒的shRNA。我们目前正在制造Lenti病毒结构，我们将用它来注射到胚胎中。已经花费了大量的时间来培训一名学生和博士后进行胚胎移植。Alaine Keebaugh和Meera Modi参加了杰克逊实验室的胚胎移植课程。因此，我们已经做好了准备，在明年真正创造出OXTR基因敲除转基因田鼠。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The objectives of this project are to develop transgenic technologies for the selective activation and inactivation of the neural systems regulating pair bonding on the genetic and cellular level. Development of these resources will have two main foci: 1) the genetic inactivation of oxytocin receptor (Oxtr) and 2) the cellular regulation of oxytocin (OT) release within transgenic lines of prairie voles. Toward the first goal we will take advantage of Cre-lox and siRNA technologies that have been used so successfully in mutant mice to regulate gene expression in a temporally regulated and cell-type specific manner. Over the past year we have screened several oxtr shRNA's to identify two that yield maximum knockdown in cell culture and in the brain. We are currently having Lenti-viral constructs being made which we will used to inject into embryos. A significant amount of time has been spent training a student and post-doctoral fellow in embryo transfer. Alaine Keebaugh and Meera Modi participated in a Jackson Laboratories Embryo transfer class. So we are well poised to actually create oxtr knockdown transgenic voles in the next year.