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Nrf2 modulates cytosolic and mitochondrial calcium signal

Alessandra Preziuso¹, Artyom Y Baev², Fozila R Rustamova²

¹Department of Biomedical Sciences and Public Health, School of Medicine, University "Politecnica delle Marche", Via Tronto 10/A, Ancona, 60126, Italy.

|May 16, 2026

Abstract:

Nrf2 is a transcription factor which regulates ∼1% of the mammalian genome and is responsible for orchestrating the cellular defense against oxidative, inflammatory and metabolic stress. Calcium (Ca²⁺) is a ubiquitous intracellular messenger which controls most cellular processes, from fertilization to cell death. Nrf2 and Ca²⁺ are involved in a large number of similar physiological processes, but it is not clear if they can regulate each other. Here, using primary co-cultures of neurons and astrocytes we asked if Nrf2 activation or deficiency alters physiological Ca²⁺ signaling and mitochondrial Ca²⁺ handling in brain cells. We found that activation of Nrf2 leads to an increase in the amplitude of Ca²⁺ peak and a faster Ca²⁺efflux in response to glutamate and ATP in neurons and astrocytes. Interestingly, Nrf2-deficient neurons and astrocytes also had higher Ca²⁺ peaks in response to glutamate and ATP, but the recovery in neurons was significantly delayed. Genetic (Keap1-knockdown) or pharmacological (ovameloxolone, RTA-408) activation of Nrf2 increases mitochondrial Ca²⁺ uptake and mitochondrial Ca²⁺ capacity, and this correlates with increased activity of the Na⁺/Ca²⁺/Li⁺ exchanger (NCLX) and inhibition of the mitochondrial permeability transition pore (mPTP). Conversely, mitochondria in neurons and astrocytes from Nrf2-knockout mice had a lower Ca²⁺ uptake, lower mitochondrial Ca²⁺ capacity and lower mitochondrial Ca²⁺efflux, making these cell vulnerable to Ca²⁺-induced cell death. Thus, Nrf2 modulates cytosolic calcium signaling and activates the mitochondrial NCLX, increasing the mitochondrial Ca²⁺ capacity, which adds another critical aspect to the multifaceted nature of Nrf2-mediated cytoprotection.

Insights

Nuclear factor erythroid 2-related factor 2 (Nrf2) influences cellular calcium (Ca2+) signaling and mitochondrial handling in brain cells. Nrf2 activation enhances Ca2+ signaling and mitochondrial capacity, offering crucial cytoprotection.

Area of Science:

Neuroscience
Cell Biology
Biochemistry

Background:

Nuclear factor erythroid 2-related factor 2 (Nrf2) is a key transcription factor regulating cellular defense mechanisms against various stresses.
Calcium (Ca2+) acts as a vital intracellular messenger controlling numerous cellular functions.
The interplay between Nrf2 and Ca2+ signaling in brain cells remains incompletely understood.

Purpose of the Study:

To investigate whether Nrf2 activation or deficiency impacts physiological Ca2+ signaling and mitochondrial Ca2+ handling in primary neuronal and astrocytic co-cultures.
To elucidate the role of Nrf2 in modulating cytosolic and mitochondrial Ca2+ dynamics in the brain.

Main Methods:

Primary co-cultures of neurons and astrocytes were utilized.
Nrf2 activation was achieved through genetic (Keap1-knockdown) and pharmacological (ovameloxolone, RTA-408) means.
Nrf2 deficiency was studied using Nrf2-knockout mouse models.
Calcium signaling and mitochondrial Ca2+ handling were assessed using various techniques.

Main Results:

Nrf2 activation increased the amplitude and accelerated the efflux of Ca2+ in response to glutamate and ATP in both neurons and astrocytes.
Nrf2-deficient cells exhibited elevated Ca2+ peaks, with delayed recovery in neurons.
Nrf2 activation enhanced mitochondrial Ca2+ uptake and capacity, correlating with increased Na+/Ca2+/Li+-exchanger (NCLX) activity and mPTP inhibition.
Nrf2 deficiency impaired mitochondrial Ca2+ handling, increasing vulnerability to Ca2+-induced cell death.

Conclusions:

Nrf2 significantly modulates cytosolic Ca2+ signaling dynamics in brain cells.
Nrf2 activation enhances mitochondrial Ca2+ buffering capacity via NCLX activation, contributing to cytoprotection.
These findings highlight a novel role for Nrf2 in regulating cellular calcium homeostasis and mitochondrial function.

Read full analysis

Keywords:

Astrocyte Calcium signal Keap1 Mitochondria Neuron Nrf2

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