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An Easily Compatible Eye-tracking System for Freely-moving Small Animals

Neuroscience Bulletin 2022年第6期38卷 661-676页

作者： Kang Huang Qin Yang Yaning Han Yulin Zhang Zhiyi Wang Liping Wang Pengfei Wei shenzhen key lab of neuropsychiatric modulation and collaborative innovation center for brain science Guang-dong Provincial Key Laboratory of Brain Connectome and BehaviorCAS Center for Excellence in Brain Science and Intelligence TechnologyBrain Cognition and Brain Disease InstituteShenzhen-Hong Kong Institute of Brain Science-Shenzhen Fundamental Research InstitutionsShenzhen Institute of Advanced TechnologyChinese Academy of SciencesShenzhen 518055China University of Chinese Academy of Seiences Bejing 100049China Harbin Institute of Technology shenzhen Shenzhen 518055.China

Measuring eye movement is a fundamental approach in cognitive science as it provides a variety of insightful parameters that reflect brain states such as visual attention and emotions.Combining eye-tracking with multimodal neural recordings or manipulation techniques is beneficial for understanding the neural substrates of cognitive function.Many commercially-available and custom-built systems have been widely applied to awake,head-fixed small animals.However,the existing eyetracking systems used in freely-moving animals are still limited in terms of their compatibility with other devices and of the algorithm used to detect eye movements.Here,we report a novel system that integrates a general-purpose,easily compatible eye-tracking hardware with a robust eye feature-detection algorithm.With ultra-light hardware and a detachable design,the system allows for more implants to be added to the animal's exposed head and has a precise synchronization module to coordinate with other neural implants.Moreover,we systematically compared the performance of existing commonly-used pupil-detection approaches,and demonstrated that the proposed adaptive pupil feature-detection algorithm allows the analysis of more complex and dynamic eye-tracking data in freemoving animals.Synchronized eye-tracking and electroencephalogram recordings,as well as algorithm validation under five noise conditions,suggested that our system is flexibly adaptable and can be combined with a wide range of neural manipulation and recording technologies.

关键词： Eye-tracking Freely-moving Head-mounted device Pupil detection Adaptive Kalman filter

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MouseVenue3D:A Markerless Three-Dimension Behavioral Tracking System for Matching Two-Photon brain Imaging in Free-Moving Mice

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Neuroscience Bulletin 2022年第3期38卷 303-317页

作者： Yaning Han Kang Huang Ke Chen Hongli Pan Furong Ju Yueyue Long Gao Gao Runlong Wu Aimin Wang Liping Wang Pengfei Wei shenzhen key laboratory of neuropsychiatric modulation and collaborative innovation center for brain science Guangdong Provincial Key Laboratory of Brain Connectome and BehaviorCAS Center for Excellence in Brain Science and Intelligence TechnologyBrain Cognition and Brain Disease InstituteShenzhen Institute of Advanced TechnologyChinese Academy of SciencesShenzhen-Hong Kong Institute of Brain ScienceShenzhen Fundamental Research InstitutionsShenzhen518055China University of Chinese Academy of sciences Beijing100049China University of Rochester RochesterNY14627USA Honam University Gwangju62399South Korea State key laboratory of Membrane Biology Institute of Molecular MedicinePeking UniversityBeijing100101China Department of Electronics Peking UniversityBeijing100871China State key laboratory of Advanced Optical Communication Systems and Networks Peking UniversityBeijing100101China

Understanding the connection between brain and behavior in animals requires precise monitoring of their behaviors in three-dimensional(3-D)space.However,there is no available three-dimensional behavior capture system that focuses on rodents.Here,we present MouseVenue3D,an automated and low-cost system for the efficient capture of 3-D skeleton trajectories in markerless rodents.We improved the most time-consuming step in 3-D behavior capturing by developing an automatic calibration module.Then,we validated this process in behavior recognition tasks,and showed that 3-D behavioral data achieved higher accuracy than 2-D data.Subsequently,MouseVenue3D was combined with fast high-resolution miniature two-photon microscopy for synchronous neural recording and behavioral tracking in the freely-moving mouse.Finally,we successfully decoded spontaneous neuronal activity from the 3-D behavior of mice.Our findings reveal that subtle,spontaneous behavior modules are strongly correlated with spontaneous neuronal activity patterns.

关键词： Computational neuroethology Behavioral and neural recording 3-D pose estimation Multi-view cameras Automatic calibration

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An Infrared Touch System for Automatic Behavior Monitoring

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Neuroscience Bulletin 2021年第6期37卷 815-830页

作者： Qingqing Liu Xing Yang Ru Song Junying Su Moxuan Luo Jinling Zhong Liping Wang shenzhen key laboratory of neuropsychiatric modulation and collaborative innovation center for brain science Guangdong Provincial Key Laboratory of Brain Connectome and BehaviorCAS Center for Excellence in Brain Science and Intelligence Technologythe Brain Cognition and Brain Disease InstituteShenzhen Institutes of Advanced TechnologyChinese Academy of SciencesShenzhen 518055China University of the Chinese Academy of sciences Beijing 100049China University of science and Technology of China Hefei 230026China

key requirements of successful animal behavior research in the laboratory are robustness,objectivity,and high throughput,which apply to both the recording and analysis of behavior.Many automatic methods of monitoring animal behavior meet these requirements.However,they usually depend on high-performing hardware and sophisticated software,which may be expensive.Here,we describe an automatic infrared behavior-monitor(AIBM)system based on an infrared touchscreen frame.Using this,animal positions can be recorded and used for further behavioral analysis by any PC supporting touch events.This system detects animal behavior in real time and gives closed-loop feedback using relatively low computing resources and simple algorithms.The AIBM system automatically records and analyzes multiple types of animal behavior in a highly efficient,unbiased,and low-cost manner.

关键词： Automatic behavior detection Elevated plus maze Two-chamber Looming Foot-shock Optogenetics Fiber photometry Heart rate and blood pressure

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modulation of Predator Cue-Evoked Tonic Immobility by Acetylcholine Released in the Basolateral Complex of the Amygdala

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Neuroscience Bulletin 2021年第11期37卷 1599-1604页

作者： Nan Liu Kang Huang Pengfei Wei Xiaolei Liu Liping Wang shenzhen key laboratory of neuropsychiatric modulation and collaborative innovation center for brain science Guangdong Provincial Key Laboratory of Brain Connectome and BehaviorCAS Center for Excellence in Brain Science and Intelligence TechnologyBrain Cognition and Brain Disease InstituteShenzhen-Hong Kong Institute of Brain Science-Shenzhen Fundamental Research InstitutionsShenzhen Institute of Advanced TechnologyChinese Academy of SciencesShenzhen518055China University of the Chinese Academy of sciences Beijing100049China

Dear Editor,Tonic immobility(TI),also termed"feigning death," is the ultimate defensive response to in escapable threats.Since some predators do not like to eat dead animals.

关键词： threat Release Ton

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Activation of parvalbumin interneurons in anterior cingulate cortex impairs observational fear

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science Bulletin 2018年第12期63卷 771-778页

作者： Chunran Zhou Zheng Zhou Yushui Han Zhuogui Lei Lei Li Quentin Montardy Xuemei Liu Fuqiang Xu Liping Wang shenzhen key laboratory of neuropsychiatric modulation and collaborative innovation center for brain science.Guangdong Provincial key laboratory of brain Connectome and Behavior CAS Center for Excellence in Brain Science and Intelligence TechnologyThe Brain Cognition and Brain Disease Institute(BCBDI).Shenzhen Institutes of Advanced TechnologyChinese Academy of SciencesShenzhen 518055China University of Chinese Academy of sciences Beijing 100049.China Southern Medical University Guangzhou 510168China center for brain science Key Laboratory of Magnetic Resonance in Biological Systems and State Key Laboratory of Magnetic Resonance and Atomic and Molecular PhysicsWuhan institute of Physics and MathematicsCAS Center for Excellence in Brain Science and Intelligence TechnologyChinese Academy of SciencesWuhan 430071China

The ability to detect conspecific's distress is crucial for animal survival.In rodent models,observational fear(OF) occurs when one animal perceives another fear related negative emotions,which may model certain behaviors caused by witnessing traumatic experiences in humans.Anterior cingulate cortex(ACC) has been showed to play a crucial role in OF.However,cellular and neural circuit basis relating to ACC governing OF is poorly understood.Here,we used Designer Receptor Exclusively Activated by a Designer Drug(DREADD) system to investigate the cell type specific circuit mechanism of ACC in OF.Firstly,inhibitory hM4D(Gi) designer receptor together with clozapine N-oxide(CNO) injection was applied to inactivate ACC neurons in the observer mice.We found that,chemogenetic inhibition of ACC resulted in a decreased freezing response in the observer mice.Next,combining PV-ires-Cre mice and Cre-dependent DREADD system,we selectively targeted the ACC parvalbumin(PV) interneurons with the excitatory hM3D(Gq) designer receptor.Activation of ACC PV interneurons following CNO injection reduced freezing response in the observer mice,while had no effect on freezing response in the demonstrator mice.Finally,monosynaptic rabies retrograde tracing revealed that ACC PV interneurons receive inputs from the mediodorsal thalamic nucleus(MD) and the ventromedial thalamic nucleus(VM),both known for their roles in OF.Taken together,these findings reveal that ACC activation is important for OF,during which PV interneurons in ACC play an important regulatory role.Abnormal function of ACC PV interneurons might contribute to the pathology of empathy-deficits related diseases,such as autism and schizophrenia.

关键词： parvalbumin 激活外皮损害动物模型 ACC 电路基础设计者

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Fabrication and modification of implantable optrode arrays for in vivo optogenetic applications

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Biophysics Reports 2018年第2期4卷 82-93页

作者： Lulu Wang Kang Huang Cheng Zhong Liping Wang Yi Lu shenzhen key lab of neuropsychiatric modulation and collaborative innovation center for brain science CAS Center for Excellence in Brain Science and Intelligence Technology The Brain Cognition and Brain Disease Institute Shenzhen Institutes of Advanced Technology Chinese Academy of Sciences shenzhen College of Advanced Technology University of Chinese Academy of Sciences

Recent advances in optogenetics have established a precisely timed and cell-specific methodology for understanding the functions of brain circuits and the mechanisms underlying neuropsychiatric disorders. However, the fabrication of optrodes, a key functional element in optogenetics, remains a great challenge. Here, we report reliable and efficient fabrication strategies for chronically implantable optrode arrays. To improve the performance of the fabricated optrode arrays, surfaces of the recording sites were modified using optimized electrochemical processes. We have also demonstrated the feasibility of using the fabricated optrode arrays to detect seizures in multiple brain regions and inhibit ictal propagation in vivo. Furthermore, the results of the histology study imply that the electrodeposition of composite conducting polymers notably alleviated the inflammatory response and improved neuronal survival at the implant/neural-tissue interface. In summary, we provide reliable and efficient strategies for the fabrication and modification of customized optrode arrays that can fulfill the requirements of in vivo optogenetic applications.

关键词： Optogenetics Optrode Neural electrode Electrodeposition Optical stimulation Electrophysiological recording

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Characterization of glutamatergic VTA neural population responses to aversive and rewarding conditioning in freely-moving mice

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science Bulletin 2019年第16期64卷 1167-1178页

作者： Quentin Montardy Zheng Zhou Zhuogui Lei Xuemei Liu Pengyu Zeng Chen Chen Yuanming Liu Paula Sanz-Leon Kang Huang Liping Wang shenzhen key lab of neuropsychiatric modulation and collaborative innovation center for brain science Guangdong Provincial Key Laboratory of Brain Connectome and BehaviorCAS Center for Excellence in Brain Science and Intelligence TechnologyThe Brain Cognition and Brain Disease Institute(BCBDI)Shenzhen Institutes of Advanced TechnologyChinese Academy of SciencesShenzhen-Hong Kong Institute of Brain Science-Shenzhen Fundamental Research InstitutionsShenzhen 518055China University of Chinese Academy of sciences Beijing 100049China School of Physics the University of SydneySydney 2006Australia Centre for Integrative brain Function the University of SydneySydney 2006Australia Department of Biomedical sciences City University of Hong KongTat Chee AvenueKowloonHong KongSAR 999077China

The Ventral Tegmental Area(VTA) is a midbrain structure known to integrate aversive and rewarding stimuli, but little is known about the role of VTA glutamatergic(VGluT2) neurons in these functions.Direct activation of VGluT2 soma evokes rewarding behaviors, while activation of their downstream projections evokes aversive behaviors. To facilitate our understanding of these conflicting properties, we recorded calcium signals from VTAVGluT2+ neurons using fiber photometry in VGluT2-cre mice to investigate how this population was recruited by aversive and rewarding stimulation, both during unconditioned and conditioned protocols. Our results revealed that, as a population, VTAVGluT2+neurons responded similarly to unconditioned-aversive and unconditioned-rewarding stimulation. During aversive and rewarding conditioning, the CS-evoked responses gradually increased across trials whilst the US-evoked response remained stable. Retrieval 24 h after conditioning, during which mice received only CS presentation, resulted in VTAVGluT2+ neurons strongly responding to CS presentation and to the expected-US but only for aversive conditioning. To help understand these differences based on VTAVGluT2+ neuronal networks, the inputs and outputs of VTAVGluT2+ neurons were investigated using Cholera Toxin B(CTB) and rabies virus. Based on our results, we propose that the divergent VTAVGluT2+ neuronal responses to aversion and reward conditioning may be partly due to the existence of VTAVGluT2+ subpopulations that are characterized by their connectivity.

关键词： Ventral Tegmental Area Aversion Reward Conditioning Behavior

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The Glutamatergic Postrhinal Cortex–Ventrolateral Orbitofrontal Cortex Pathway Regulates Spatial Memory Retrieval

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Neuroscience Bulletin 2019年第3期35卷 447-460页

作者： Xinyang Qi Zhanhong Jeff Du Lin Zhu Xuemei Liu Hua Xu Zheng Zhou Cheng Zhong Shijiang Li Liping Wang Zhijun Zhang Department of Neurology Affiliated ZhongDa HospitalInstitute of NeuropsychiatrySchool of MedicineSoutheast UniversityNanjing 210009China shenzhen key lab of neuropsychiatric modulation and collaborative innovation center for brain science Chinese Academy of Sciences (CAS) Center for Excellence in Brain Science and Intelligence Technologythe Brain Cognition and Brain Disease Institute for Collaboration Research of the Shenzhen Institutes of Advanced Technology at the CAS and the McGovern Institute at Massachusetts Institute of TechnologyShenzhen 518055China Department of Biophysics Medical College of WisconsinMilwaukeeWI53226USA

A deficit in spatial memory has been taken as an early predictor of Alzheimer’s disease(AD) or mild cognitive impairment(MCI). The uncinate fasciculus(UF) is a long-range white-matter tract that connects the anterior temporal lobe with the orbitofrontal cortex(OFC)in primates. Previous studies have shown that the UF impairment associated with spatial memory deficits may be an important pathological change in aging and AD, but its exact role in spatial memory is not well understood. The pathway arising from the postrhinal cortex(POR) and projecting to the ventrolateral orbitofrontal cortex(vlOFC)performs most of the functions of the UF in rodents.Although the literature suggests an association between spatial memory and the regions connected by the POR–vlOFC pathway, the function of the pathway in spatialmemory is relatively unknown. To further illuminate the function of the UF in spatial memory, we dissected the POR–vlOFC pathway in mice. We determined that the POR–vlOFC pathway is a glutamatergic structure, and that glutamatergic neurons in the POR regulate spatial memory retrieval. We also demonstrated that the POR–vlOFC pathway specifically transmits spatial information to participate in memory retrieval. These findings provide a deeper understanding of UF function and dysfunction related to disorders of memory, as in MCI and AD.

关键词： Spatial memory Postrhinal cortex Ventrolateral orbitofrontal cortex Mild cognitive impairment Alzheimer's disease

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Rabies Virus Pseudotyped with CVS-N2C Glycoprotein as a Powerful Tool for Retrograde Neuronal Network Tracing

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Neuroscience Bulletin 2020年第3期36卷 202-216页

作者： Xutao Zhu Kunzhang Lin Qing Liu Xinpei Yue Huijie Mi Xiaoping Huang Xiaobin He Ruiqi Wu Danhao Zheng Dong Wei Liangliang Jia Weilin Wang Anne Manyande Jie Wang Zhijian Zhang Fuqiang Xu shenzhen key lab of neuropsychiatric modulation and collaborative innovation center for brain science Guangdong Provincial Key Laboratory of Brain Connectome and BehaviorCAS Center for Excellence in Brain Science and Intelligence TechnologyBrain Cognition and Brain Disease Institute(BCBDI)Shenzhen Institutes of Advanced TechnologyChinese Academy of SciencesShenzhen-Hong Kong Institute of Brain Science-Shenzhen Fundamental Research InstitutionsShenzhen 518055China State key laboratory of Magnetic Resonance and Atomic and Molecular Physics Key Laboratory of Magnetic Resonance in Biological SystemsWuhan Center for Magnetic ResonanceWuhan Institute of Physics and MathematicsChinese Academy of SciencesWuhan 430071China Wuhan National laboratory for Optoelectronics Huazhong University of Science and TechnologyWuhan 430071China College of Life sciences Wuhan UniversityWuhan 430071China School of Human and Social sciences University of West LondonLondonUK University of the Chinese Academy of sciences Beijing 100049China

Efficient viral vectors for mapping and manipulating long-projection neuronal circuits are crucial in structural and functional studies of the brain. The SAD strain rabies virus with the glycoprotein gene deleted pseudotyped with the N2 C glycoprotein(SAD-RV(DG)-N2 C(G)) shows strong neuro-tropism in cell culture, but its in vivo efficiency for retrograde gene transduction and neuro-tropism have not been systematically characterized.We compared these features in different mouse brain regions for SAD-RV-N2 C(G) and two other widely-used retrograde tracers, SAD-RV(DG)-B19(G) and r AAV2-retro. We found that SAD-RV(DG)-N2 C(G) enhanced the infection efficiency of long-projecting neurons by^10 times but with very similar neuro-tropism, compared with SAD-RV(DG)-B19(G). On the other hand, SAD-RV(DG)-N2 C(G) had an infection efficiency comparable with r AAV2-retro, but a more restricted diffusion range, and broader tropism to different types and regions of longprojecting neuronal populations. These results demonstrate that SAD-RV(DG)-N2 C(G) can serve as an effective retrograde vector for studying neuronal circuits.

关键词： Viral vector N2C glycoprotein Neuronal circuits Retrograde tracing

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Structure and function of superior colliculus-thalamicamygdala subcoritcal pathway and its role in visually evoked innate fear procesing

Structure and function of superior colliculus-thalamicamygda...

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中国神经科学学会第十二届全国学术会议

作者： Peng-Fei WEI Nan LIU Zhen Ren Wen-Hao Liu Li-Ping WANG shenzhen key lab of neuropsychiatric modulation and collaborative innovation center for brain science CAS Center for Excellence in Brain Science and Intelligence Technology the Brain Cognition and Brain Disease Institute （BCBDI） for Collaboration Research of SIAT at CAS and the McGovern Institute at MIT Shenzhen Institutes of Advanced Technology Chinese Academy of Sciences

A fundamental question in the fields of neuroscience and cognition science is: how the brain works for sensing,processing and handling the information from external stimulus, especially for emotion-laden stimuli. Individuals have its inherent ability to evaluate the biological significance of affective stimuli through a modular, automatically brain mechanism. More specifically, detection and reaction of an unexpected salient stimuli, especially of potential danger or emergency in the natural environment, is crucial for human and animal’s survival. The standard hypothesis suggests that the processing of threating stimuli initiate by a subcortical pathway which largely bypass cortical information processing. A potential threat object, such as an approaching predator, can be detected by the midbrain superior colliculus, and the visual pulvinar Abstracts the key features of threat signal then directly send to amygdala to generate the fear responses. However, there is still lack evidence of detail neural circuitry mechanisms to support this idea and the controversy exists about the precise relay pathway that might connect the superior colliculus with the amygdala. To address this question, we established a behavioral paradigm to evoke an unlearned fearful freezing in mice by presenting a looming stimulus from overhead. By utilizing optogenetic, multi-areas electrophysiology virus based tracing techniques, we performed a careful characterization of a subcortical pathway superior colliculus-lateral nucleus of the posterior thalamus（pulvinar in rodents）-lateral amygdala. For the first time, we found the unrevealed role of this subcortical pathway on processing visually evoked innate defensive responses, and this pathway very specifically encodes a stereotyped unlearned fear-like freezing behavior. Importantly, we also showed that activity in lateral amygdala is necessary for the display of looming triggered defensive behaviors and is correlated with neuronal activity in the SC. Furthermore, the activation of the innate f

关键词： Optogenetics Subcortical visual pathway Innate fear

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