A Whole-Brain Cell-Type-Specific Sparse Neuron Labeling Method and Its Application in a Shank3 Autistic Mouse Model
A Whole-Brain Cell-Type-Specific Sparse Neuron Labeling Method and Its Application in a Shank3 Autistic Mouse Model
复制标题
全脑细胞类型特异性稀疏神经元标记方法及其在 Shank3 自闭症小鼠模型中的应用。
DOI：
10.3389/fncel.2020.00145
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发表时间：
2020-06-05
影响因子：
5.3
通讯作者：
Wang, Wenting
中科院分区：
文献类型：
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作者：
Chen, Di;Ren, Keke;Wang, Wenting
单个神经元作为脑的基本单位，由细胞体和突起（包括树突和轴突）组成。即使是同一类型的神经元也会表现出各种微妙的过程特征，以适应不同的神经回路。不同类型的神经元细胞在大脑中形成复杂的回路。因此，需要详细的神经元形态学来了解正常的神经元功能和病理机制，例如自闭症中发生的那些。在这里，我们开发了一种策略，在整个大脑中稀疏标记相同类型的神经元，并在自闭症动物模型-Shank 3基因敲除（KO）小鼠中测试其应用。为了实现这一点，我们设计了一种腺相关病毒（AAV），表达Cre重组酶依赖的常规和膜靶向增强型绿色荧光蛋白（EGFP）下的人突触蛋白1启动子，并验证了它在几个Cre转基因小鼠。我们可以稀疏标记的投射神经元在多个脑区的后眼注射的病毒到CaMKII α-Cre小鼠。然后，我们用这种方法对Shank 3 KO小鼠的投射神经元进行了形态学分析。我们发现，与同窝对照小鼠相比，Shank 3 KO小鼠与CaMKII α-Cre小鼠在纹状体、皮质和海马中投射神经元的树突复杂性和树突棘变化存在差异。通过将这种方法与各种Cre小鼠系与小鼠疾病模型相结合，我们可以筛选整个大脑中不同类型神经元的形态学特征，这将有助于我们了解特定细胞类型的神经元不仅在自闭症谱系障碍（ASD）小鼠模型中的确切作用，而且在其他精神疾病小鼠模型中。
Single neurons, as the basic unit of the brain, consist of a cell body and processes, including dendrites and axons. Even neurons of the same type show various subtle process characteristics to fit into the diverse neural circuits. Different cell types of neurons form complicated circuits in the brain. Therefore, detailed neuronal morphology is required to understand normal neuronal function and pathological mechanisms, such as those that occur in autism. Here, we developed a strategy to sparsely label the same type of neurons throughout the whole brain and tested its application in an autistic animal model-Shank3knockout (KO) mice. To achieve this, we designed an adeno-associated virus (AAV) that expresses Cre recombinase-dependent regular and membrane-targeted enhanced green fluorescent protein (EGFP) under a human synapsin 1 promoter and verified it in several Cre transgenic mice. We could sparsely label the projection neurons in multiple brain areas by retro-ocular injection of the virus into CaMKII alpha-Cre mice. Then, we analyzed the morphology of the projection neurons inShank3KO mice with this method. We found differential dendritic complexity and dendritic spine changes in projection neurons inShank3KO mice crossed with CaMKII alpha-Cre mice compared with littermate control mice in the striatum, cortex, and hippocampus. By combining this method with various Cre mouse lines crossed with mouse models of disease, we can screen the morphological traits of distinct types of neurons throughout the whole brain that will help us to understand the exact role of the specific cell types of neurons not only in autism spectrum disorder (ASD) mouse models but also in other psychiatric disorder mouse models.