支配初级视皮层的丘脑背外侧核神经元亚群的小鼠全脑输入图谱构建

doi:10.3969/j.issn.1674-8115.2021.06.001

上海交通大学学报(医学版) ›› 2021, Vol. 41 ›› Issue (6): 701-709.doi: 10.3969/j.issn.1674-8115.2021.06.001

支配初级视皮层的丘脑背外侧核神经元亚群的小鼠全脑输入图谱构建

王丽昭¹^,²(), 马国芬¹^,²(), 刘燕梅¹^,², 王彦婕¹^,², 王紫玥¹^,², 陈兆南¹^,², 张思宇¹^,²(), 徐天乐¹^,²()

^1.上海交通大学医学院附属上海儿童医学中心儿童脑科学中心，上海 200127
^2.上海交通大学基础医学院解剖学与生理学系，上海 200025

出版日期:2021-06-28 发布日期:2021-06-29
通讯作者: 张思宇,徐天乐 E-mail:wanglizhao@shsmu.edu.cn;zhang_siyu@sjtu.edu.cn;zhang_siyu@shsmu.edu.cn;xu-happiness@shsmu.edu.cn
作者简介:王丽昭（1982—），女，助理实验师，硕士；电子信箱：wanglizhao@shsmu.edu.cn|王丽昭（1982—），女，助理实验师，硕士；电子信箱：wanglizhao@shsmu.edu.cn
基金资助:
国家自然科学基金(31900711);上海交通大学医学院高水平地方高校创新团队(SSMU-ZDCX20181100);上海市科学技术委员会基础研究重点项目(18JC1420302);上海市市级科技重大专项(2018SHZDZX05)

Whole-brain inputs mapping to the projection neurons in laterodorsal thalamic nucleus innervating primary visual cortex in mice

Li-zhao WANG¹^,²(), Guo-fen MA¹^,²(), Yan-mei LIU¹^,², Yan-jie WANG¹^,², Zi-yue WANG¹^,², Zhao-nan CHEN¹^,², Si-yu ZHANG¹^,²(), Tian-le XU¹^,²()

^1.Center for Brain Science, Shanghai Children's Medical Center, Shanghai Jiao Tong University School of Medicine, Shanghai 200127, China
^2.Department of Anatomy and Physiology, Shanghai Jiao Tong University College of Basic Medical Sciences, Shanghai 200025, China

Online:2021-06-28 Published:2021-06-29
Contact: Si-yu ZHANG,Tian-le XU E-mail:wanglizhao@shsmu.edu.cn;zhang_siyu@sjtu.edu.cn;zhang_siyu@shsmu.edu.cn;xu-happiness@shsmu.edu.cn
Supported by:
National Natural Science Foundation of China(31900711);Innovative Research Team of High-Level Local Universities in Shanghai(SSMU-ZDCX20181100);Basic Research Project of Science and Technology Commission of Shanghai Municipality(18JC1420302);Key Project of Science and Technology Commission of Shanghai Municipality(2018SHZDZX05)

摘要/Abstract

摘要：

目的·在小鼠全脑范围定量分析支配初级视皮层的丘脑背外侧核神经元亚群的直接输入神经元。方法·向C57BL/6J雄性小鼠初级视皮层注射表达小麦胚芽凝集素和Cre重组酶融合蛋白的腺相关病毒，同时向丘脑背外侧核注射Cre重组酶依赖的分别表达禽类肉瘤/白血病病毒包膜蛋白受体TVA和狂犬病毒糖蛋白的假性狂犬病毒辅助腺相关病毒。28 d后，向初级视皮层注射假性狂犬病毒。待假性狂犬病毒携带的绿色荧光蛋白在丘脑背外侧核神经元亚群的上游输入神经元中充分表达后，即7 d后，收集小鼠脑部样本，并制作冰冻切片。荧光显微镜下观察支配初级视皮层的丘脑背外侧核神经元亚群的输入神经元在脑内的分布情况，并使用高通量解剖数据分析软件进行定量分析。结果·荧光显微镜观察及定量分析结果发现，支配初级视觉皮层的丘脑背外侧核神经元亚群的输入神经元在皮层、间脑和中脑均有分布，分别占全脑绿色荧光标记神经元总数的（58.1±4.8）%、（23.2±0.9）%和（15.3±4.0）%。在皮层区域，输入神经元分布最多的区域是躯体运动皮层、视皮层和躯体感觉皮层；在前额叶皮层和中间联合皮层也有大量输入神经元分布；在感觉皮层中，输入神经元的胞体主要分布在第5层和第6层。结论·支配初级视觉皮层的丘脑背外侧核神经元亚群的输入神经元在全脑分布广泛，其中多数位于皮层区。

关键词: 丘脑背外侧核, 初级视皮层, 全脑输入图谱, 小鼠, 假性狂犬病毒

Abstract:

Objective·To analyze the whole-brain inputs to the projection neurons in the lateral dorsal nucleus (LD) of thalamus innervating primary visual cortex (V1) in mice.

Methods·The adeno-associated virus (AAV) vectors expressing the fusion protein of wheat germ agglutinin and Cre recombinase (Cre) were injected into V1 of C57BL/6J male mice, and the AAV vectors with Cre-inducible expression of avian sarcoma/leukosis virus envelope glycoprotein receptor TVA and rabies glycoprotein were injected into LD, respectively. Twenty-eight days later, the pseudotyped rabies virus (RV) was injected into V1. After the full expression of RV-mediated green fluorescent protein in the infected input neurons of LD, i.e., 7 d later, brain samples were collected and frozen sections were made. The whole-brain distribution of RV-labeled neurons were observed under fluorescence microscope, and quantitative analysis was performed by using high-throughput anatomical data analysis software.

Results·Through fluorescence microscope and quantitative analysis, the whole-brain map showed that the inputs of LD neurons innervating V1 mainly came from cortex, middle brain and inter brain, accounting for (58.1±4.8)%, (23.2±0.9)% and (15.3±4.0)% of the total number of GFP-labeled neurons in the whole brain, respectively. In the cortex, the areas with the most input neurons were somatomotor area, visual area and somatosensory area. A large number of input neurons were also distributed in prefrontal cortex and medial commissural cortex. In the sensory cortex, the cell bodies of the inputs were mainly distributed in the fifth and sixth layers.

Conclusion·The direct inputs of LD neuron subgroups innervating V1 in mice are widely distributed throughout the brain, most of which are located in the cortex.

Key words: lateral dorsal nucleus of thalamus, primary visual cortex, whole-brain inputs, mouse, pseudotyped rabies virus

中图分类号:

Q422

王丽昭, 马国芬, 刘燕梅, 王彦婕, 王紫玥, 陈兆南, 张思宇, 徐天乐. 支配初级视皮层的丘脑背外侧核神经元亚群的小鼠全脑输入图谱构建[J]. 上海交通大学学报(医学版), 2021, 41(6): 701-709.

Li-zhao WANG, Guo-fen MA, Yan-mei LIU, Yan-jie WANG, Zi-yue WANG, Zhao-nan CHEN, Si-yu ZHANG, Tian-le XU. Whole-brain inputs mapping to the projection neurons in laterodorsal thalamic nucleus innervating primary visual cortex in mice[J]. JOURNAL OF SHANGHAI JIAOTONG UNIVERSITY (MEDICAL SCIENCE), 2021, 41(6): 701-709.

图/表 5

图1 标记支配初级视皮层的丘脑背外侧核神经元全脑输入神经元的病毒联合使用策略Note:A. Viruses used in the experiment. B. Upper panel, the procedure of RV-mediated trans-synaptic retrograde tracing. Lower panel, the illustration for injection sites. C. The time schedule of viral vector injection.

Fig 1 Strategy for joint viral labeling of the whole-brain inputs to LD neurons innervating V1

图2 病毒注射位点及荧光蛋白在各级神经元的表达情况Note:A. The injection site of AAV-EF1a-mCherry-IRES-WGA-Cre. B. The injection site of AAV-DIO-TVA-mCherry and AAV-DIO-RG. C. PRV-labeled input neurons to LD neurons innervating V1. D. Start neurons in the LD of thamalus.

Fig 2 Viral vector injection sites and the expression of fluorescence proteins in the neurons

图3 全脑范围内假性狂犬病毒逆向示踪标记的输入神经元Note:Input neurons (green) in the cortical areas (A), interbrain (B), and midbrain (C).

Fig 3 Whole-brain distribution of pseudorabies virus-labeled input neurons

图4 全脑范围内各脑区假性狂犬病毒逆向示踪标记的输入神经元百分比Note:Percentages of labeled input neurons in cortical areas (A), interbrain (B), and midbrain (C).

Fig 4 Percentages of pseudorabies virus-labeled input neurons in the whole brain

图5 假性狂犬病毒逆向标记的神经元在各感觉皮层中的分布

Fig 5 Laminar distribution of pseudorabies virus-labeled input neurons in the sensory cortices

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支配初级视皮层的丘脑背外侧核神经元亚群的小鼠全脑输入图谱构建

Whole-brain inputs mapping to the projection neurons in laterodorsal thalamic nucleus innervating primary visual cortex in mice

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