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Related Experiment Video

Updated: Oct 4, 2025

Imaging Mitochondrial Ca2+ Uptake in Astrocytes and Neurons using Genetically Encoded Ca2+ Indicators GECIs
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Imaging Mitochondrial Ca2+ Uptake in Astrocytes and Neurons using Genetically Encoded Ca2+ Indicators GECIs

Published on: January 22, 2022

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Imaging Mitochondrial Ca2+ Uptake in Astrocytes and Neurons using Genetically Encoded Ca2+ Indicators (GECIs).

Nannan Zhang1, Zhe Zhang2, Ilker Ozden3

  • 1Dalton Cardiovascular Research Center, University of Missouri-Columbia.

Journal of Visualized Experiments : Jove
|February 7, 2022
PubMed
Summary

Researchers developed a novel method for imaging mitochondrial calcium (Ca2+) in brain cells. This technique allows for cell-specific visualization of Ca2+ dynamics in astrocytes and neurons, aiding the study of neurological diseases.

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Last Updated: Oct 4, 2025

Imaging Mitochondrial Ca2+ Uptake in Astrocytes and Neurons using Genetically Encoded Ca2+ Indicators GECIs
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Analyses of Mitochondrial Calcium Influx in Isolated Mitochondria and Cultured Cells
08:29

Analyses of Mitochondrial Calcium Influx in Isolated Mitochondria and Cultured Cells

Published on: April 27, 2018

14.0K

Area of Science:

  • Neuroscience
  • Cell Biology
  • Biochemistry

Background:

  • Mitochondrial calcium (Ca2+) is crucial for cellular functions like energy metabolism and signal transduction.
  • Dysregulation of mitochondrial Ca2+ is implicated in neurodegenerative diseases and cell death.
  • Existing methods lack cell-type specificity and targeted mitochondrial visualization in astrocytes and neurons.

Purpose of the Study:

  • To develop a cell-type specific, mitochondria-targeting molecular approach for imaging mitochondrial Ca2+.
  • To enable in vitro and in vivo visualization of mitochondrial Ca2+ dynamics in astrocytes and neurons.
  • To facilitate the study of astrocyte-neuron interactions and Ca2+ signaling.

Main Methods:

  • Constructed DNA plasmids encoding mitochondria-targeting genetically encoded Ca2+ indicators (GECIs) GCaMP5G/6s.
  • Utilized astrocyte-specific (gfaABC1D) and neuron-specific (CaMKII) promoters for targeted expression.
  • Employed plasmid transfection for in vitro imaging and adeno-associated viral vectors (AAVs) for in vivo mouse brain imaging.

Main Results:

  • Successfully expressed GCaMP5G/6s in mitochondria of cultured astrocytes and neurons.
  • Achieved in vivo expression of GCaMP5G/6s in astrocyte and neuron mitochondria within the mouse brain.
  • Demonstrated a viable method for imaging mitochondrial Ca2+ dynamics in specific brain cell types.

Conclusions:

  • The developed approach enables precise imaging of mitochondrial Ca2+ in astrocytes and neurons.
  • This tool is valuable for investigating the interplay between cytosolic and mitochondrial Ca2+ signaling.
  • The method supports research into astrocyte-neuron communication and its role in neurological health and disease.