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

Dhyani, Vaibhav; Swain, Sarpras; Gupta, Rishikesh Kumar; Saxena, Abha; Singh, Ranjana; Giri, Lopamudra

doi:10.1007/978-1-0716-1107-4_5

Cited by 3 publications

(3 citation statements)

References 49 publications

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“…To compress the rich information observed in these time-resolved profiles into a means of comparing conditions, we performed feature-based k -means clustering . Based on the DB-index values, four clusters were observed in this data set (Supplementary Figure S7A–D).…”

Section: Resultsmentioning

confidence: 99%

Three-Dimensional Tracking of Intracellular Calcium and Redox State during Real-Time Control in a Hypoxic Gradient in Microglia Culture: Comparison of the Channel Blocker and Reoxygenation under Ischemic Shock

Dhyani

Kumar

Manne

et al. 2023

ACS Chem. Neurosci.

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Real-time three-dimensional (3-D) imaging is crucial for quantifying correlations among various molecules under acute ischemic stroke. Insights into such correlations may be decisive in selecting molecules capable of providing a protective effect within a shorter period. The major bottleneck is maintaining the cultures under severely hypoxic conditions while simultaneously 3-D imaging intracellular organelles with a microscope. Moreover, comparing the protective effect of drugs and reoxygenation remains challenging. To address this, we propose a novel workflow for the induction of gas-environment-based hypoxia in the HMC-3 cells along with 3-D imaging using laser-scanning-confocal microscopy. The imaging framework is complemented with a pipeline for quantifying time-lapse videos and cell-state classification. First, we show an imaging-based assessment of the in vitro model for hypoxia using a steep gradient in O 2 with time. Second, we demonstrate the correlation between mitochondrial superoxide production and cytosolic calcium under acute hypoxia. We then test the efficacy of an L-type calcium channel blocker, compare the results with reoxygenation, and show that the blocker alleviates hypoxic conditions in terms of cytosolic calcium and viability within an acute window of one hour. Furthermore, we show that the drug reduces the expression of oxidative stress markers (HIF1A and OXR1) within the same time window. In the future, this model can also be used to investigate drug toxicity and efficacy under ischemic conditions.

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Section: Resultsmentioning

confidence: 99%

Three-Dimensional Tracking of Intracellular Calcium and Redox State during Real-Time Control in a Hypoxic Gradient in Microglia Culture: Comparison of the Channel Blocker and Reoxygenation under Ischemic Shock

Dhyani

Kumar

Manne

et al. 2023

ACS Chem. Neurosci.

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“…Eagle's minimal essential medium was used to culture the cells. On the fourth day of culture, astrocyte growth was inhibited by adding 10 μM cytosine arabinoside (Dhyani et al, 2021; Figure 1).…”

Section: Methodsmentioning

confidence: 99%

“…Since the characterization of Ca 2+ signals from dissociated neurons remains challenging, several attempts have been made to develop statistical formulations that quantify cell‐to‐cell heterogeneity via clustering and distribution fitting (Dhyani et al, 2021; Mölter et al, 2018; Swain et al, 2018). Despite the use of machine learning methods for functional clustering, effectively grouping heterogeneous responses remains challenging (Muldoon et al, 2013).…”

Section: Introductionmentioning

confidence: 99%

A computational model to uncover the biophysical underpinnings of neural firing heterogeneity in dissociated hippocampal cultures

Dhyani,

George,

Gare

et al. 2023

Hippocampus

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Calcium (Ca2+) imaging reveals a variety of correlated firing in cultures of dissociated hippocampal neurons, pinpointing the non‐synaptic paracrine release of glutamate as a possible mediator for such firing patterns, although the biophysical underpinnings remain unknown. An intriguing possibility is that extracellular glutamate could bind metabotropic receptors linked with inositol trisphosphate (IP3) mediated release of Ca2+ from the endoplasmic reticulum of individual neurons, thereby modulating neural activity in combination with sarco/endoplasmic reticulum Ca2+ transport ATPase (SERCA) and voltage‐gated Ca2+ channels (VGCC). However, the possibility that such release may occur in different neuronal compartments and can be inherently stochastic poses challenges in the characterization of such interplay between various Ca2+ channels. Here we deploy biophysical modeling in association with Monte Carlo parameter sampling to characterize such interplay and successfully predict experimentally observed Ca2+ patterns. The results show that the neurotransmitter level at the plasma membrane is the extrinsic source of heterogeneity in somatic Ca2+ transients. Our analysis, in particular, identifies the origin of such heterogeneity to an intrinsic differentiation of hippocampal neurons in terms of multiple cellular properties pertaining to intracellular Ca2+ signaling, such as VGCC, IP3 receptor, and SERCA expression. In the future, the biophysical model and parameter estimation approach used in this study can be upgraded to predict the response of a system of interconnected neurons.

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Microstate analysis of GABA_B and mGluR Mediated Modulation of Calcium Spiking in Hippocampal Neurons

Sehanobish,

Dhyani,

Gare

et al. 2023

2023 11th International IEEE/EMBS Conference on Neural Engineering (NER)

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

Cited by 3 publications

References 49 publications

Three-Dimensional Tracking of Intracellular Calcium and Redox State during Real-Time Control in a Hypoxic Gradient in Microglia Culture: Comparison of the Channel Blocker and Reoxygenation under Ischemic Shock

Three-Dimensional Tracking of Intracellular Calcium and Redox State during Real-Time Control in a Hypoxic Gradient in Microglia Culture: Comparison of the Channel Blocker and Reoxygenation under Ischemic Shock

A computational model to uncover the biophysical underpinnings of neural firing heterogeneity in dissociated hippocampal cultures

Microstate analysis of GABA_B and mGluR Mediated Modulation of Calcium Spiking in Hippocampal Neurons

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

Cited by 3 publications

References 49 publications

Three-Dimensional Tracking of Intracellular Calcium and Redox State during Real-Time Control in a Hypoxic Gradient in Microglia Culture: Comparison of the Channel Blocker and Reoxygenation under Ischemic Shock

Three-Dimensional Tracking of Intracellular Calcium and Redox State during Real-Time Control in a Hypoxic Gradient in Microglia Culture: Comparison of the Channel Blocker and Reoxygenation under Ischemic Shock

A computational model to uncover the biophysical underpinnings of neural firing heterogeneity in dissociated hippocampal cultures

Microstate analysis of GABAB and mGluR Mediated Modulation of Calcium Spiking in Hippocampal Neurons

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Microstate analysis of GABA_B and mGluR Mediated Modulation of Calcium Spiking in Hippocampal Neurons