Assaying Circuit Specific Regulation of Adult Hippocampal Neural Precursor Cells

Luis J. Quintanilla; Chia-Yu Yeh; Hechen Bao; Christina Catavero; Juan Song

doi:10.3791/59237

Regulação específica do circuito de assaying de células precursoras neurais hipocampais adultas

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Neuroscience

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JoVE Journal Neuroscience

Assaying Circuit Specific Regulation of Adult Hippocampal Neural Precursor Cells

Regulação específica do circuito de assaying de células precursoras neurais hipocampais adultas

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08:52 min

July 24, 2019

DOI: 10.3791/59237-v

Luis J. Quintanilla^1,2,3, Chia-Yu Yeh^1,2, Hechen Bao^1,2, Christina Catavero^1,2,3, Juan Song^1,2,3

¹Department of Pharmacology,University of North Carolina Chapel Hill, ²Neuroscience Center,University of North Carolina Chapel Hill, ³Neuroscience Curriculum,University of North Carolina Chapel Hill

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Overview

This protocol describes a method for analyzing the behavior of adult neural stem/progenitor cells when subjected to chemogenetic manipulation of specific local neural circuits. The approach aims to elucidate how neural circuit stimulation or inhibition impacts adult neurogenesis.

Key Study Components

Area of Science

Neuroscience
Neurogenesis
Cell Biology

Background

Investigation of adult neural stem cell proliferation.
Understanding the regulation by specific neural circuits.
Role of neurotransmitters in adult neurogenesis.
Micro-manipulation techniques to reduce stress in animal models.

Purpose of Study

To evaluate the effect of targeted neural circuit manipulation on adult neurogenesis.
To assess the proliferation of neural stem cells.
To determine the functionality of specific neurotransmitter-releasing cell types.

Methods Used

Use of tissue sections and immunohistochemistry for analysis.
Adult neural stem/progenitor cells manipulated via chemogenetic techniques.
No multiomics workflow is mentioned in the study.
Key steps include tissue preparation, staining, and microscopy imaging.
Quantification of cell density and neurogenesis using imaging software.

Main Results

Confirmed the effectiveness of targeted neural circuit manipulation on neural stem cell proliferation.
Demonstrated fluorescent labeling of specific cell types involved in neurogenesis.
Proliferation rates of nestin-positive cells and their morphological characteristics analyzed.
Effective quantification methods established for evaluating neurogenesis across different experimental conditions.

Conclusions

The study illustrates the impact of local circuit activity on the regulation of neural stem cells.
The methodology enhances the understanding of neurogenic processes in the adult brain.
Possible implications for therapeutic strategies targeting neurogenesis in neurological diseases.

Frequently Asked Questions

What are the advantages of this protocol?

This protocol allows for precise manipulation of neural circuits while minimizing stress in animal subjects, enhancing the study’s reliability.

How is the neural stem cell proliferation measured?

Proliferation is assessed through fluorescence microscopy, identifying cells labeled with thymidine analogs and counting their density.

What type of tissue is used in this study?

Adult brain tissue sections are used to analyze the behavior of neural stem/progenitor cells under specific conditions.

How can this method be adapted for other studies?

The methodology can be tailored to investigate other neurotransmitter systems or different neural pathways associated with neurogenesis.

What are potential limitations of this approach?

The results may be influenced by the specific conditions set during circuit manipulation or the accessibility of target cells in the tissue sections.

What data outcomes can be obtained from this protocol?

Outcomes include cell density measurements, identification of proliferating progenitor cells, and insights into the regulatory effects of neurotransmitters on neurogenesis.

O objetivo deste protocolo é descrever uma aproximação para analisar o comportamento de pilhas neurais adultas da haste/progenitor em resposta à manipulação chemogenetic de um circuito neural local específico.

Este protocolo pode responder como vários circuitos no cérebro regulam neurogênese adulta e especificamente como estimular ou inibir circuitos neurais afetam a proliferação de células-tronco neurais adultas. A vantagem dessa técnica é que ela é capaz de atingir especificamente circuitos neurais desejados, bem como reduzir a quantidade de estresse introduzido aos animais. Este método pode fornecer uma visão de como diferentes circuitos cerebrais regulam a neurogênese adulta, em particular, como tipos de células específicas que liberam certos neurotransmissores regulam a proliferação de células-tronco neurais adultas.

Para iniciar este procedimento, coloque as seções teciduais em PBS e monte cinco a oito seções em um slide carregado positivamente em ordem serial de anterior para posterior. Deixe as seções de tecido secarem à temperatura ambiente por dois a cinco minutos e adere completamente aos slides. Em seguida, prepare o tampão de citrato em um recipiente.

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