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

Regulación específica del circuito de ensayo de células precursoras neuronales hipocampales adultas

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Neuroscience

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JoVE Journal Neuroscience

Assaying Circuit Specific Regulation of Adult Hippocampal Neural Precursor Cells

Regulación específica del circuito de ensayo de células precursoras neuronales hipocampales 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.

El objetivo de este protocolo es describir un enfoque para analizar el comportamiento de las células madre/progenitoras neurales adultas en respuesta a la manipulación quimiogenética de un circuito neural local específico.

Este protocolo puede responder cómo varios circuitos en el cerebro regulan la neurogénesis adulta y específicamente cómo estimular o inhibir los circuitos neuronales afectan la proliferación de células madre neurales adultas. La ventaja de esta técnica es que es capaz de apuntar específicamente a los circuitos neuronales deseados, así como reducir la cantidad de estrés introducido a los animales. Este método puede proporcionar información sobre cómo diferentes circuitos cerebrales regulan la neurogénesis adulta, en particular, cómo los tipos de células específicas que liberan ciertos neurotransmisores regulan la proliferación de células madre neurales adultas.

Para comenzar este procedimiento, coloque las secciones de tejido en PBS y monte de cinco a ocho secciones en una diapositiva cargada positivamente en orden de serie de anterior a posterior. Deje que las secciones del tejido se sequen a temperatura ambiente durante dos a cinco minutos y se adhieran completamente a los portaobjetos. A continuación, prepare el búfer de citrato en un contenedor.

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