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。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 表示子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子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，用于在遗传和细胞水平上选择性激活和失活调节配对结合的神经系统。这些资源的开发将有两个主要焦点：1）催产素受体（Oxtr）的遗传失活和2）草原田鼠转基因系中催产素（OT）释放的细胞调节。为了实现第一个目标，我们将利用Cre-lox和siRNA技术，这些技术已经成功地用于突变小鼠，以时间调节和细胞类型特异性的方式调节基因表达。在过去的一年中，我们筛选了几个oxtr shRNA，以确定两个在细胞培养和大脑中产生最大敲低的基因。 我们目前正在制造慢病毒结构，我们将用它来注射到胚胎中。 花了大量的时间培训一名学生和博士后胚胎移植研究员。 Alaine Keebaugh和Meera Modi参加了杰克逊实验室的胚胎移植课程。 所以我们已经做好准备在明年制造出oxtr基因敲除的转基因田鼠。

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.