Comparative analyses of transgene expression patterns after intra‐striatal injections of rAAV2‐retro in rats and rhesus monkeys: A light and electron microscopic study,European Journal of Neuroscience

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Comparative analyses of transgene expression patterns after intra‐striatal injections of rAAV2‐retro in rats and rhesus monkeys: A light and electron microscopic study
European Journal of Neuroscience ( IF 2.7 ) Pub Date : 2020-10-28 , DOI: 10.1111/ejn.15027
Daniel L. Albaugh _{1,

2} , Yoland Smith _{1,

2,

3} , Adriana Galvan _{1,

2,

3}

Affiliation

Retrogradely‐transducing viral vectors are versatile tools for anatomical and functional interrogations of neural circuits. These vectors can be applied in nonhuman primates (NHPs), powerful model species for neuroscientific studies with limited genetic tractability, but limited data are available regarding the tropism and transgene expression patterns of such viruses after injections in NHP brains. Consequently, NHP researchers must often rely on related data available from other species for experimental planning. To evaluate the suitability of rAAV2‐retro in the NHP basal ganglia, we studied the transgene expression patterns at the light and electron microscope level after injections of rAAV2‐retro vector encoding the opsin Jaws conjugated to a green fluorescent protein (GFP) in the putamen of rhesus macaques. For inter‐species comparison, we injected the same vector in the rat dorsal striatum. In both species, GFP expression was observed in numerous cortical and subcortical regions with known striatal projections. However, important inter‐species differences in pathway transduction were seen, including labeling of the intralaminar thalamostriatal projection in rats, but not monkeys. Electron microscopic ultrastructural observations within the basal ganglia revealed GFP labeling in both postsynaptic dendrites and presynaptic axonal terminals; the latter likely derived from anterograde transgene transport in neurons that project to the striatum, and from collaterals of these neurons. Our results suggest that certain neural pathways may be refractory to transduction by retrograde vectors in a species‐specific manner, highlighting the need for caution when determining the suitability of a retrograde vector for NHP studies based solely on rodent data.

中文翻译：

大鼠和恒河猴纹状体内注射rAAV2-retro后转基因表达模式的比较分析：光学和电子显微镜研究

逆行转导的病毒载体是用于神经回路的解剖和功能询问的多功能工具。这些载体可用于非人类灵长类动物（NHPs），这是用于神经科学研究的功能强大的模型物种，具有有限的遗传可操作性，但有关注射入NHP脑后此类病毒的向性和转基因表达模式的数据有限。因此，NHP研究人员必须经常依靠其他物种的相关数据进行实验计划。为了评估rAAV2-retro在NHP基底神经节中的适用性，我们研究了在视核细胞中注射编码视蛋白钳与绿色荧光蛋白（GFP）结合的视蛋白钳的rAAV2-retro载体后在光镜和电子显微镜下的转基因表达模式恒河猴。为了进行种间比较，我们在大鼠背侧纹状体中注射了相同的载体。在这两个物种中，在许多具有已知纹状体投射的皮质和皮质下区域均观察到了GFP表达。但是，在通路转导中发现了重要的种间差异，包括标记了大鼠（而非猴子）的椎板内丘脑投射。基底节内的电子显微超微结构观察显示，在突触后树突和突触前轴突末端都有GFP标记。后者可能源自投射到纹状体的神经元的顺行转基因转运，以及这些神经元的侧支。我们的结果表明，某些神经通路可能难以逆转载体以物种特异性方式进行转导，

更新日期：2020-12-31

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