Real-time Live-cell Flow Cytometry to Investigate Calcium Influx, Pore Formation, and Phagocytosis by P2X7 Receptors in Adult Neural Progenitor Cells

作者： Hannah C. Leeson1,2, Tailoi Chan-Ling3,4, Michael D. Lovelace3,5,6, Jeremy D. Brownlie7, Michael W. Weible II*1,4,7, Ben J. Gu*8 1Griffith Institute for Drug Discovery,Griffith University, 2Australian Institute for Bioengineering and Nanotechnology,University of Queensland, 3Discipline of Anatomy and Histology, School of Medical Science,University of Sydney, 4Bosch Institute,University of Sydney, 5Applied Neurosciences Program, Peter Duncan Neurosciences Research Unit,St. Vincent's Centre for Applied Medical Research, 6School of Medical Sciences,The University of New South Wales (UNSW) Medicine, Sydney, New South Wales, 7School of Environment and Science,Griffith University, Brisbane, Queensland, 8Florey Institute of Neuroscience and Mental Health,University of Melbourne

简介：

Overview

This article presents a protocol for assessing P2X7 receptor functions in adult neural progenitor cells using real-time flow cytometry. The method enables the quantification of calcium influx, transmembrane pore formation, and phagocytosis.

Key Study Components

Area of Science

• Neuroscience
• Cell Biology
• Receptor Biology

Background

• P2X7 receptors exhibit versatile functions based on agonist concentration.
• Understanding P2X7 receptor modulation in neural progenitor cells is crucial for neuroscience research.
• This protocol is adaptable to various cell types.
• Rapid and reproducible methods are essential for effective research.

Purpose of Study

• To investigate the three major functions of the P2X7 receptor.
• To provide a quantifiable method for studying receptor activity.
• To enhance understanding of receptor modulation in neural progenitor cells.

Methods Used

• Isolation of neural progenitor cells from adult mice.
• Flow cytometry for real-time analysis of receptor functions.
• Calcium influx measurement using calcium indicator dye.
• Assessment of transmembrane pore formation and phagocytosis.

Main Results

• Successful quantification of P2X7 receptor functions in live cultures.
• Demonstrated the versatility of P2X7 receptors in different physiological contexts.
• Provided a clear methodology for future studies on receptor modulation.

Conclusions

• The protocol allows for effective study of P2X7 receptor functions.
• Real-time flow cytometry is a valuable tool for neuroscience research.
• Adaptability of the method makes it suitable for various cell types.

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