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

Dhyani, Vaibhav and Swain, Sarpras and Gupta, Rishikesh Kumar and Saxena, Abha and Singh, Ranjana and Giri, Lopamudra (2021) Role of Metabotropic Glutamate Receptors (mGluRs) in the Regulation of Cellular Calcium Signaling: Theory, Protocols, and Data Analysis. Neuromethods, 164. pp. 81-115. ISSN 0893-2336

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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.

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IITH Creators:

IITH Creators	ORCiD
Dhyani, Vaibhav	UNSPECIFIED
Swain, Sarpras	UNSPECIFIED
Saxena, Abha	UNSPECIFIED
Giri, Lopamudra	http://orcid.org/0000-0002-2352-7919

Item Type:

Article

Uncontrolled Keywords:

animal cell; animal tissue; automation; calcium signaling; confocal microscopy; controlled study; correlation analysis; data analysis; data clustering; density functional theory; hippocampus; methodology; nerve cell culture; nerve cell network; nonhuman; priority journal; probability; protein function; rat; regulatory mechanism; signal transduction

Subjects:

Chemical Engineering

Divisions:

Department of Chemical Engineering

Depositing User:

. LibTrainee 2021

Date Deposited:

09 Jul 2021 05:13

Last Modified:

09 Jul 2021 05:13

URI:

http://raiithold.iith.ac.in/id/eprint/8184