In Vivo Calcium Imaging of Granule Cells in the Dentate Gyrus of Hippocampus in Mice

Shanshan Han; Ning Ding; Ce Li; Peng Yuan

doi:10.3791/66916

In Vivo: Calcium Imaging of Granule cells in the dentate gyrus of hippocampus in mice(생쥐의 해마...

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Neuroscience

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JoVE Journal Neuroscience

In Vivo Calcium Imaging of Granule Cells in the Dentate Gyrus of Hippocampus in Mice

In Vivo: Calcium Imaging of Granule cells in the dentate gyrus of hippocampus in mice(생쥐의 해마 치열계에 있는 과립 세포의 In vivo 칼슘 이미징)

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07:00 min

August 2, 2024

DOI: 10.3791/66916-v

Shanshan Han¹, Ning Ding¹, Ce Li¹, Peng Yuan¹

¹Department of Rehabilitation Medicine, Huashan Hospital, State Key Laboratory of Medical Neurobiology, Institute for Translational Brain Research, MOE Frontiers Center for Brain Science, MOE Innovative Center for New Drug Development of Immune Inflammatory Diseases,Fudan University

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Overview

This study outlines a protocol for in vivo calcium imaging of granule cells in the dentate gyrus of the hippocampus in freely moving mice. Focused on investigating neuroactivities, the protocol combines virus injection and GRIN lens implantation in a single surgical procedure. This novel approach aims to enhance data quality and accuracy in understanding the neural coding and spatial representations in cognitive functions.

Key Study Components

Area of Science

Neuroimaging
Neuroscience
Cognition

Background

The dentate gyrus plays a crucial role in learning and memory.
Calcium imaging is vital for studying neuronal activities in real-time.
Traditional methods separate virus injection and lens implantation, which can lead to inaccuracies.
In vivo imaging allows for the observation of neural dynamics during cognitive tasks.

Purpose of Study

To improve the efficiency and accuracy of in vivo calcium imaging.
To facilitate the study of neuronal activity correlating with cognitive functions.
To understand the mechanisms underlying spatial representations in the dentate gyrus.

Methods Used

In vivo calcium imaging was performed using a miniature microscope.
The biological model included granule cells in the dentate gyrus of freely moving mice.
The protocol integrates virus injection with GRIN lens implantation to streamline the procedure.
Critical steps involve precise surgical techniques to ensure accurate placements.
Data analysis includes motion correction and single-cell signal extraction from imaging data.

Main Results

Successful imaging revealed varying numbers of active cells based on visibility of blood vessels.
Data showed individual cell responses that illuminate the coding properties of the dentate gyrus.
The integration of surgical steps enhances the overall quality of collected data.
Different active cell counts were associated with imaging field quality.

Conclusions

The study demonstrates an effective method for in vivo calcium imaging in freely moving mice.
This approach facilitates improved examination of neuroactivities related to cognitive processing.
Results have implications for understanding cognitive mechanisms and potential neurological disorders.

Frequently Asked Questions

What are the advantages of the combined surgical approach?

Combining virus injection and GRIN lens implantation minimizes wait times and improves accuracy, leading to better imaging quality.

How is the biological model implemented in the study?

The model involves freely moving mice with granule cells in the dentate gyrus being studied for neuroactivities using calcium imaging.

What types of data are obtained from calcium imaging?

The imaging provides insights into the neurodynamic patterns associated with cognition and allows for the identification of active neurons.

How can this method be adapted for other research?

This protocol can be modified for different neuroanatomical regions or species, enhancing its applicability in various neuroscience research contexts.

Are there any limitations to the protocol?

Key considerations include ensuring proper placement of the GRIN lens and maintaining clear visibility during imaging, which may require advanced surgical skills.

What implications do the findings have for understanding brain function?

The study's findings contribute to a deeper understanding of how spatial representations and neural coding underpin cognitive functions in the brain.

해마의 치상이랑(dentate gyrus)은 학습과 기억에서 필수적이고 뚜렷한 기능을 수행합니다. 이 프로토콜은 자유롭게 움직이는 마우스의 치상이랑에 있는 과립 세포의 생체 내 칼슘 이미징을 위한 일련의 강력하고 효율적인 절차를 설명합니다.

글쎄요, 이 프로토콜에서는 소형 현미경을 사용하여 해마의 치상이랑의 신경 활동을 조사합니다. 우리는 다양한 질병 조건에서 뉴런 코딩 및 공간 표현을 조사하는 것을 목표로 합니다. in vivo dentate gyrus 연구의 주요 과제는 고품질 칼슘 이미징 데이터를 얻는 것, 바이러스 발현이 만족스러운지, 그래디언트가 적절한 위치에 이식되었는지 여부와 관련이 있다고 생각합니다.

그리고 이와 같은 세부 사항, 전체 수술은 연구의 성공에 매우 중요합니다. 전통적인 칼슘 이미징 수술에서 바이러스 주입과 GRIN 렌즈 이식은 일반적으로 별도의 수술로 수행됩니다. 우리 프로토콜에서는 이 두 단계를 단일 절차로 결합하여 대기 시간을 줄일 뿐만 아니라 GRIN 렌즈의 정확한 배치를 보장합니다.

우리 연구실은 다양한 현미경 기술을 사용하여 인지 기능의 기저에 있는 신경 역학을 연구합니다. 우리는 인지를 지원하는 신경 활동 패턴을 이해하기를 희망합니다. 그리고 이론적 모델링과 인과 관계 조작을 결합하여 뇌의 정보 흐름과 신경 계산 과정을 해부하기를 희망합니다.

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