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Role of Metabotropic Glutamate Receptors (mGluRs) in the Regulation of Cellular Calcium Signaling: Theory, Protocols, and Data Analysis

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Metabotropic Glutamate Receptor Technologies

Part of the book series: Neuromethods ((NM,volume 164))

Abstract

Glutamate is the primary fast excitatory neurotransmitter which is responsible for neuronal communication, memory formation, and learning. It activates metabotropic glutamate receptors (mGluRs), a family of G protein-coupled receptors, which lead to oscillations in cytosolic calcium (Ca2+) through a variety of signaling cascades. Mechanisms of these cascades and their modulation by drugs are well understood, but there are limited methodological guidelines available for systematic study of Ca2+-spiking behavior and quantification of spiking information. Neurons are known to exhibit an extensive variety of Ca2+-spiking behavior from bursting spikes to sustained plateau, thus constituting the oscillations into high-dimensional time-series data, comprising of heterogeneous features (amplitude, frequency, inter-spike interval, etc.). This chapter addresses the signaling pathways involved in mGluR-mediated Ca2+-spiking along with a system of nonlinear ordinary differential equation used for explaining glutamate-induced-Ca2+-responses. We also discuss the modulatory effect of mGluRs on other channels and receptors involved in Ca2+-signaling. The chapter specifically focuses on the details of the imaging systems and parameters that can be used for high-resolution-Ca2+-imaging for monitoring neural activity. Here, we provide a detailed workflow of algorithms for various data analysis tools including clustering, measuring correlations, and probability density function (PDF) fitting for analyzing the heterogeneous data obtained from mGluR-mediated-Ca2+-spiking in primary hippocampal neurons. Moreover, we provide sample codes and MATLAB functions that can be used for automated analysis of large-scale-Ca2+-spiking data. Since there is a high level of heterogeneity present in dissociated cultured neurons, we provide a protocol for grouping the data and random sampling to perform the statistical analysis using clustering and PDF fitting.

All authors contributed to the conception and design of the chapter. VD, SS, RKG and LG planned the complete work. All authors wrote various sections of the manuscript. VD contributed to the theory of mGluRs and prepared the schematic diagrams and figures. SS contributed to the protocol writing for clustering and correlation analysis. RKG provided the details of experiments and the imaging setup. AS provided the theory and analysis of PDF fitting. RS performed analysis of early and delayed DHPG response. LG performed the imaging experiments. All authors read and approved the final manuscript.

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Acknowledgments

We thank Dr. Steven Mennerick for providing the hippocampal neurons and performing the experiments with dissociated neurons. We also thank Dr. Narsimhan Gautam for performing imaging assays using confocal microscopy at Washington University School of Medicine, St. Louis.We thank Vani Kalyanraman and Dr. Soumya Jana for their valuable discusssions.

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Authors and Affiliations

  1. Bioimaging and Data Analysis Laboratory, Indian Institute of Technology, Hyderabad, India

    Vaibhav Dhyani, Sarpras Swain, Abha Saxena, Ranjana Singh & Lopamudra Giri

  2. Laboratory of Neurodegeneration, International Institute of Molecular and Cell Biology in Warsaw, Warsaw, Poland

    Rishikesh Kumar Gupta

  3. School of Molecular Medicine, Medical University of Warsaw, Warsaw, Poland

    Rishikesh Kumar Gupta

Authors
  1. Vaibhav Dhyani
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  2. Sarpras Swain
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  3. Rishikesh Kumar Gupta
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  4. Abha Saxena
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  5. Ranjana Singh
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  6. Lopamudra Giri
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Corresponding author

Correspondence to Lopamudra Giri .

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Editors and Affiliations

  1. Psychology Department, Arizona State University, Tempe, AZ, USA

    M Foster Olive

  2. Phoenix Children’s Hospital, Barrow Neurological Institute, Phoenix, AZ, USA

    Brian T. Burrows

  3. Psychology Department, Arizona State University, Tempe, AZ, USA

    Jonna M. Leyrer-Jackson

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Dhyani, V., Swain, S., Gupta, R.K., Saxena, A., Singh, R., Giri, L. (2021). Role of Metabotropic Glutamate Receptors (mGluRs) in the Regulation of Cellular Calcium Signaling: Theory, Protocols, and Data Analysis. In: Olive, M.F., Burrows, B.T., Leyrer-Jackson, J.M. (eds) Metabotropic Glutamate Receptor Technologies. Neuromethods, vol 164. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1107-4_5

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  • DOI: https://doi.org/10.1007/978-1-0716-1107-4_5

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1106-7

  • Online ISBN: 978-1-0716-1107-4

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