Laboratory of Cellular and Molecular Neurotherapies, Center for Neuroscience Research, Clinical Neurosciences Department, Lausanne University Hospital, Switzerland.
Laboratory of Cellular and Molecular Neurotherapies, Center for Neuroscience Research, Clinical Neurosciences Department, Lausanne University Hospital, Switzerland.
J Neurosci Methods. 2021 Apr 15;354:109105. doi: 10.1016/j.jneumeth.2021.109105. Epub 2021 Feb 27.
PV mice--> combined with AAV-FLEX vectors allowed efficient and specific targeting of PV interneurons in the striatum. However, diffusion of viral particles to the globus pallidus caused massive transduction of PV projection neurons and subsequent anterograde transport of the transgene product to the subthalamic nucleus and the substantia nigra pars reticulata. Different AAV serotypes (1 and 9) and promoters (CBA and human synapsin) were evaluated. The combination of AAV1, a moderate expression level (human synapsin promoter) and a precise adjustment of the stereotaxic coordinates in the anterior and dorsolateral part of the striatum were necessary to avoid transduction of PV GP projection neurons. Even in the absence of direct transduction due to diffusion of viral particles, GP PV projection neurons could be retrogradely transduced via their terminals present in the dorsal striatum. However, in the absence of diffusion, GP-Str PV projection neurons were poorly or not transduced suggesting that retrograde transduction did not significantly impair the selective targeting of striatal PV neurons. Finally, a prominent reduction of the number of striatal PV interneurons (about 50 %) was evidenced in the presence of the Cre recombinase suggesting that functional effects of AAV-mediated transgene expression in PV striatal interneurons in PV mice should be analyzed with caution.
PV 小鼠--结合 AAV-FLEX 载体，可实现纹状体中 PV 中间神经元的高效、特异性靶向。然而，病毒颗粒向苍白球的扩散导致大量的 PV 投射神经元被转导，并随后将转基因产物顺行转运到丘脑底核和黑质网状部。评估了不同的 AAV 血清型（1 和 9）和启动子（CBA 和人突触素）。为了避免 PV-GP 投射神经元的转导，需要结合 AAV1、适度的表达水平（人突触素启动子）和在前纹状体的前侧和背外侧部分进行精确的立体定位调整。即使由于病毒颗粒的扩散而没有直接转导，通过存在于背侧纹状体中的末端，也可以逆行转导 GP-PV 投射神经元。然而，在没有扩散的情况下，GP-Str PV 投射神经元被转导的情况较差或没有，这表明逆行转导并没有显著损害纹状体 PV 神经元的选择性靶向。最后，在 Cre 重组酶存在的情况下，纹状体 PV 中间神经元的数量明显减少（约 50%），这表明在 PV 小鼠的纹状体 PV 中间神经元中，AAV 介导的转基因表达的功能影响应谨慎分析。
