Kakava-Georgiadou Nefeli, Zwartkruis Maria M, Bullich-Vilarrubias Clara, Luijendijk Mieneke C M, Garner Keith M, van der Plasse Geoffrey, Adan Roger A H
Division of Neuroscience, Department of Translational Neuroscience, Brain Center Rudolf Magnus, University Medical Center Utrecht, Utrecht, Netherlands.
Master's Program Neuroscience and Cognition, Utrecht University, Utrecht, Netherlands.
Front Mol Neurosci. 2019 Feb 28;12:49. doi: 10.3389/fnmol.2019.00049. eCollection 2019.
Development of tools to manipulate activity of specific neurons is important for dissecting the function of neural circuits. Viral vectors and conditional transgenic animal lines that target recombinases to specific cells facilitate the successful manipulation and recording of specific subsets of neurons. So far, it has been possible to target neuronal subtypes within a certain brain region based on transcriptional control regions from a gene selectively expressed in those cells or based upon its projections. Nevertheless, there are only a few tools available that combine this and target a neuronal subtype within a projection. We tested a viral vector system, consisting of a canine adenovirus type 2 expressing a Cre-dependent Flp recombinase (CavFlexFlp) and an adeno-associated viral (AAV) vector expressing a Flp-dependent cDNA, which targets neurons in a subtype- and projection-specific manner. As proof of principle we targeted expression of a Designer Receptor Exclusively Activated by Designer Drugs (DREADD) to the dopamine neurons of the mesolimbic projection, which allows the transient activation of neurons by the ligand Clozapine-N-Oxide (CNO). We validated that the system specifically targets dopamine neurons and that chemogenetic activation of these neurons induces an increase in locomotor activity. We thus validated a valuable tool that allows neuronal activation in a projection- and subtype-specific manner.
开发用于操纵特定神经元活动的工具对于剖析神经回路的功能至关重要。将重组酶靶向特定细胞的病毒载体和条件转基因动物品系有助于成功操纵和记录特定的神经元亚群。到目前为止，基于在这些细胞中选择性表达的基因的转录控制区域或基于其投射，已经能够在特定脑区内靶向神经元亚型。然而，将这两者结合并在投射内靶向神经元亚型的工具只有少数几种。我们测试了一种病毒载体系统，该系统由表达Cre依赖性Flp重组酶的犬2型腺病毒（CavFlexFlp）和表达Flp依赖性cDNA的腺相关病毒（AAV）载体组成，该系统以亚型和投射特异性方式靶向神经元。作为原理验证，我们将仅由设计药物激活的设计受体（DREADD）的表达靶向中脑边缘投射的多巴胺神经元，这使得神经元能够被配体氯氮平 - N - 氧化物（CNO）瞬时激活。我们验证了该系统特异性靶向多巴胺神经元，并且这些神经元的化学遗传激活会导致运动活性增加。因此，我们验证了一种有价值的工具，该工具能够以投射和亚型特异性方式激活神经元。
