Connectivity and function of inhibitory neurons in the primate visual cortex

总结 本行政补充申请的目标和具体目的不变 关于母R 01奖励。在此，我们要求额外的资金， 在原项目范围内的费用。在最初的应用程序中，我们建议使用 特异性感染生长抑素（SOM）和小清蛋白（PV）抑制性神经元的病毒载体 在非人类灵长类动物的大脑皮层中。这些病毒载体是由我们的 合作者德州大学奥斯汀分校的Zemelman博士（该项目前两年的研究人员）， Dimidschstein（布罗德研究所）和Horwitz博士（华盛顿大学）。在前两年的 在项目中，我们测试并验证了我们的合作者产生的3种PV特异性病毒。 Dimidschstein博士的病毒对PV神经元的特异性最高。然而不幸的是， Dimidschstein博士最近离开了学术界，不再能够为我们提供这方面的信息。 vector及其变体。因此，我们开始与戈登博士合作， Fishell（哈佛）和Gabrielle Pouchelon博士（冷泉港），以生成 Dimidschstein PV特异性病毒，表达执行TRIO所需的基因 在该项目的目标1中提出的实验中，追踪PV电池的解剖输入。此外，委员会认为， 我们正在与私营公司（Genewiz和Addgene）和宾夕法尼亚大学病毒载体核心（U-Penn Viral Vector Core）合作， 产生这种携带兴奋和抑制基因的PV特异性病毒的变体， 视蛋白;这些是解决项目目标3所必需的。这些新方法需要 我们无法用奖励的资金支付的重大额外费用。请求的 成本增加将使我们能够使用向量来解决项目的部分Aim 1和Aim 3， >90%的特异性，通过我们在项目最初几年进行的研究验证。我们将 向社区提供额外资金产生的病媒。

SUMMARY The goals and specific aims of the present Administrative Supplement application are unchanged with respect to the parent R01 award. Here we request additional funds for unanticipated expenses within the original scope of the project. In the original application we proposed to use viral vectors that specifically infect somatostatin (SOM) and parvalbumin (PV) inhibitory neurons in non-human primate cortex. These viral vectors were generated and provided by our collaborators Dr Zemelman at UT Austin (a subcontract PI in the first two years of the project), Dr Dimidschstein (Broad Institute), and Dr Horwitz (Univ. of Washington). In the first two years of the project, we tested and validated the 3 types of PV-specific viruses generated by our collaborators. Dr Dimidschstein’s virus showed the highest specificity for PV neurons. Unfortunately, however, Dr Dimidschstein has recently left academia and is no longer in a position to provide us with this vector and its variants at no cost. We have, therefore, started a collaboration with Drs Gordon Fishell (Harvard) and Dr. Gabrielle Pouchelon (Cold Spring Harbor) to generate variants of the Dimidschstein PV-specific virus that express the necessary genes to perform the TRIO experiments proposed in Aim 1 of the project, to trace anatomical inputs to PV cells. Moreover, we are working with private companies (Genewiz and Addgene) and the U Penn Viral Vector Core to generate variants of this PV-specific virus carrying the genes for excitatory and inhibitory opsins; these are necessary to address Aim 3 of the project. These new approaches require significant additional expenses which we cannot cover with the funds awarded. The requested cost increases would allow us to address part of Aim1 and Aim3 of the project using vectors with >90% specificity as validated by our studies performed in the initial years of the project. We will make accessible to the community the vectors generated with the additional funds.