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Feedback Regulation of Calcium Concentration

Feedback Regulation of Calcium Concentration

Calcium is an essential signaling molecule required for various cellular functions. Calcium pumps and ion channels on cell and organellar membranes, such as those on the endoplasmic reticulum (ER), regulate calcium concentrations inside the cell. They remain closed, keeping the cytosolic calcium levels low at a resting state.
Various transmembrane receptors, such as G protein-coupled receptors (GPCRs), elicit a response to extracellular signals by increasing cytosolic calcium. Activated GPCRs...

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Fluorescent Calcium Imaging and Subsequent In Situ Hybridization for Neuronal Precursor Characterization in Xenopus laevis

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Calcium Oscillation Frequency Is a Potential Functional Complex Physiological Relevance Indicator for a

Amish Asthana, Kenneth Ndyabawe, Dayrin Mendez

ACS Biomaterials Science & Engineering

|January 19, 2021

Summary

New 3D neural models show physiologically relevant responses compared to 2D cultures. Calcium oscillation frequency in 3D models closely mimics in vivo data, suggesting it as a key indicator for neural drug screening.

Keywords:

3D culture HTS calcium oscillations caveolae cell-based assay neural scaffold spheroid

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Area of Science:

Neuroscience
Biomedical Engineering
Drug Discovery

Background:

In vitro drug screening for neural function relies on 2D cultures, which often exaggerate in vivo responses.
Developing physiologically relevant 3D neural models is crucial for accurate drug screening.
Complex physiologically relevant (CPR) outcomes are needed to evaluate 3D culture fidelity.

Purpose of the Study:

To evaluate calcium oscillation frequency as a functional CPR indicator in 2D versus 3D neural cell cultures.
To compare the physiological relevance of different 3D neural culture formats.

Main Methods:

SHSY5Y cells were cultured in both 2D and 3D formats (spheroids and microporous polymer scaffolds).
Fluo-4-based calcium fluorescence imaging was employed to monitor neural activity.
Automated image processing quantified calcium oscillation frequency.

Main Results:

Calcium oscillation frequency was significantly upregulated in 2D cultures.
Calcium oscillation frequency in spheroid and scaffold-based 3D models was comparable to in vivo measurements.
This suggests calcium oscillation frequency is a reliable indicator of physiological relevance.

Conclusions:

3D neural models, particularly spheroids and scaffold-based cultures, offer greater physiological relevance than traditional 2D cultures.
Calcium oscillation frequency serves as a promising functional CPR indicator for neural cultures.
These findings advance the development of more accurate in vitro models for neural drug discovery.