利用VSV标记具有特定投射的特异类型神经元的精细结构

Labeling Fine Morphology of Neurons with Specific Cell Types and Specific Projection Using Vesicular Stomatitis Virus

神经元是构成神经系统的基本单位之一,对其精细形态结构的研究是了解神经网络构筑和信息处理方式的基础,然而目前缺乏能够标记具有特定投射特征的特异类型神经元精细结构的有效方法.在糖蛋白基因缺失的水泡性口炎病毒(VSV)毒株基础上突变其核蛋白,我们获得了重组病毒VSV-(35)G-NR7A-EGFP,并发现该病毒在一定窗口期内可实现神经元形态的快速高亮度标记;我们进一步在此基础上构建了VSV-(35)G-EnvA-NR7A-EGFP病毒,并基于特定的转基因动物及辅助病毒rAAV-EF1α-Dio-Bfp-Tva,通过控制注射位点,分别实现了有NAc-LH投射特异性的D2R神经元和VTA-NAc投射特异性的多巴胺神经元的标记,展示了一种可用于稀疏、高亮地标记具有特定投射特征的特异类型神经元精细结构的新方法.

Neuron is the basic unit of nervous system.Studying the fine morphology of neurons is crucial for understanding the structure of neural network and the manner of neural information processing.At present,effective methods to visualize fine morphology of neurons with specific cell types and specific projection are still lacking.In this paper,a recombination vesicular stomatitis virus(VSV)was designed,with nucleoprotein gene mutated and glycoprotein gene deleted.The virus,VSV-?G-NR7A-EGFP,was capable of highlighted morphology of the labelled neurons in a short time.With specific transgenic animals and a helper virus rAAV-EF1α-Dio-Bfp-Tva,it was demonstrated that,a further constructed virus VSV-?G-EnvA-NR7A-EGFP could sparsely and specifically label D2R neurons with nucleus accumbens-lateral hypothalamus(NAc-LH)projection and dopamine neurons with ventral tegmental area(VTA)-NAc projection.