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

Imagem de cálcio in vivo de células granulares no giro dentado do hipocampo em camundongos

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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

Imagem de cálcio in vivo de células granulares no giro dentado do hipocampo em camundongos

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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.

O giro dentado do hipocampo desempenha funções essenciais e distintas na aprendizagem e na memória. Este protocolo descreve um conjunto de procedimentos robustos e eficientes para imagens de cálcio in vivo de células granulares no giro dentado em camundongos em movimento livre.

Bem, neste protocolo, usamos um microscópio em miniatura para investigar as atividades neurais no giro dentado do hipocampo. Nosso objetivo é investigar a codificação de neurônios e as representações espaciais durante as diferentes condições de doença. Acredito que o principal desafio de estudar o giro dentado in vivo envolve a obtenção de dados de imagem de cálcio de alta qualidade, se a expressão do vírus é satisfatória, se o gradiente é implantado no local adequado.

E os detalhes como esses, toda a cirurgia são de importância crítica para o sucesso do estudo. Na cirurgia tradicional de imagem de cálcio, a injeção do vírus e o implante da lente GRIN são normalmente realizados como operações separadas. Em nosso protocolo, combinamos essas duas etapas em um único procedimento, o que não apenas reduz o tempo de espera, mas também garante o posicionamento preciso da lente GRIN.

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