Double Virus Vector Infection to the Prefrontal Network of the Macaque Brain.
Double Virus Vector Infection to the Prefrontal Network of the Macaque Brain.
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DOI：
10.1371/journal.pone.0132825
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Sakagami M
中科院分区：
文献类型：
--
作者：
Oguchi M;Okajima M;Tanaka S;Koizumi M;Kikusui T;Ichihara N;Kato S;Kobayashi K;Sakagami M
为了准确理解大脑前额叶网络中高级认知功能的实现，需要光遗传学和药物遗传学方法来操纵特定神经通路的信号传递。然而，这些方法的应用大多局限于灵长类动物以外的动物，而灵长类动物是研究高级认知功能的最佳动物模型。在本研究中，我们采用双病毒载体感染猕猴大脑前额叶网络的方法。这使我们能够在不使用种系遗传操作的情况下，将特定结构表达到构成目标通路的特定神经元中。双重感染技术利用两个不同的病毒载体在两个单突触连接的区域。一种是局部感染投射神经元细胞体的载体（局部载体），另一种是从同一投射神经元的轴突末端逆行感染的载体（逆行载体）。逆行载体包含编码Cre重组酶的序列，而局部载体包含编码报告蛋白（mCherry）的“creon”FLEX双絮凝序列。因此，mCherry只能在这些载体双重感染的投射神经元中表达。我们将这种方法应用于两只猕猴，并针对前额叶网络中的两条不同通路：从外侧前额叶皮层到尾状核的通路和从外侧前额叶皮层到额叶视野的通路。结果，在所有四个被注射的大脑半球的外侧前额叶皮层中都观察到麦克切利阳性细胞，这表明双重病毒载体转染在猕猴大脑前额叶网络中是可行的。
To precisely understand how higher cognitive functions are implemented in the prefrontal network of the brain, optogenetic and pharmacogenetic methods to manipulate the signal transmission of a specific neural pathway are required. The application of these methods, however, has been mostly restricted to animals other than the primate, which is the best animal model to investigate higher cognitive functions. In this study, we used a double viral vector infection method in the prefrontal network of the macaque brain. This enabled us to express specific constructs into specific neurons that constitute a target pathway without use of germline genetic manipulation. The double-infection technique utilizes two different virus vectors in two monosynaptically connected areas. One is a vector which can locally infect cell bodies of projection neurons (local vector) and the other can retrogradely infect from axon terminals of the same projection neurons (retrograde vector). The retrograde vector incorporates the sequence which encodes Cre recombinase and the local vector incorporates the “Cre-On” FLEX double-floxed sequence in which a reporter protein (mCherry) was encoded. mCherry thus came to be expressed only in doubly infected projection neurons with these vectors. We applied this method to two macaque monkeys and targeted two different pathways in the prefrontal network: The pathway from the lateral prefrontal cortex to the caudate nucleus and the pathway from the lateral prefrontal cortex to the frontal eye field. As a result, mCherry-positive cells were observed in the lateral prefrontal cortex in all of the four injected hemispheres, indicating that the double virus vector transfection is workable in the prefrontal network of the macaque brain.