MLKL overexpression leads to Ca2+ and metabolic dyshomeostasis in a neuronal cell model

doi:10.1016/j.ceca.2024.102854

. 2024 May:119:102854.

doi: 10.1016/j.ceca.2024.102854. Epub 2024 Feb 6.

MLKL overexpression leads to Ca²⁺ and metabolic dyshomeostasis in a neuronal cell model

Sathyaseelan S Deepa¹, Nidheesh Thadathil², Jorge Corral³, Sabira Mohammed⁴, Sophia Pham³, Hadyn Rose³, Michael T Kinter⁵, Arlan Richardson⁶, Carlos Manlio Díaz-García⁷

Affiliations

¹ Department of Biochemistry and Physiology, University of Oklahoma Health Sciences Center, OK, USA; Center for Geroscience and Healthy Brain Aging, University of Oklahoma Health Sciences Center, OK, USA; Stephenson Cancer Center, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA. Electronic address: deepa-sathyaseelan@ouhsc.edu.
² Department of Biochemistry and Physiology, University of Oklahoma Health Sciences Center, OK, USA.
³ Department of Biochemistry and Physiology, University of Oklahoma Health Sciences Center, OK, USA; Center for Geroscience and Healthy Brain Aging, University of Oklahoma Health Sciences Center, OK, USA.
⁴ Department of Biochemistry and Physiology, University of Oklahoma Health Sciences Center, OK, USA; Center for Geroscience and Healthy Brain Aging, University of Oklahoma Health Sciences Center, OK, USA; Stephenson Cancer Center, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA.
⁵ Aging & Metabolism Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, USA.
⁶ Department of Biochemistry and Physiology, University of Oklahoma Health Sciences Center, OK, USA; Center for Geroscience and Healthy Brain Aging, University of Oklahoma Health Sciences Center, OK, USA; Stephenson Cancer Center, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA; Oklahoma City VA Medical Center, Oklahoma City, OK, USA.
⁷ Department of Biochemistry and Physiology, University of Oklahoma Health Sciences Center, OK, USA; Center for Geroscience and Healthy Brain Aging, University of Oklahoma Health Sciences Center, OK, USA; Harold Hamm Diabetes Center, University of Oklahoma Health Sciences Center, OK, USA. Electronic address: carlosmanlio-diazgarcia@ouhsc.edu.

PMID: 38430790
PMCID: PMC10990772 (available on 2025-05-01)
DOI: 10.1016/j.ceca.2024.102854

MLKL overexpression leads to Ca²⁺ and metabolic dyshomeostasis in a neuronal cell model

Sathyaseelan S Deepa et al. Cell Calcium. 2024 May.

. 2024 May:119:102854.

doi: 10.1016/j.ceca.2024.102854. Epub 2024 Feb 6.

Authors

Sathyaseelan S Deepa¹, Nidheesh Thadathil², Jorge Corral³, Sabira Mohammed⁴, Sophia Pham³, Hadyn Rose³, Michael T Kinter⁵, Arlan Richardson⁶, Carlos Manlio Díaz-García⁷

Affiliations

¹ Department of Biochemistry and Physiology, University of Oklahoma Health Sciences Center, OK, USA; Center for Geroscience and Healthy Brain Aging, University of Oklahoma Health Sciences Center, OK, USA; Stephenson Cancer Center, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA. Electronic address: deepa-sathyaseelan@ouhsc.edu.
² Department of Biochemistry and Physiology, University of Oklahoma Health Sciences Center, OK, USA.
³ Department of Biochemistry and Physiology, University of Oklahoma Health Sciences Center, OK, USA; Center for Geroscience and Healthy Brain Aging, University of Oklahoma Health Sciences Center, OK, USA.
⁴ Department of Biochemistry and Physiology, University of Oklahoma Health Sciences Center, OK, USA; Center for Geroscience and Healthy Brain Aging, University of Oklahoma Health Sciences Center, OK, USA; Stephenson Cancer Center, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA.
⁵ Aging & Metabolism Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, USA.
⁶ Department of Biochemistry and Physiology, University of Oklahoma Health Sciences Center, OK, USA; Center for Geroscience and Healthy Brain Aging, University of Oklahoma Health Sciences Center, OK, USA; Stephenson Cancer Center, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA; Oklahoma City VA Medical Center, Oklahoma City, OK, USA.
⁷ Department of Biochemistry and Physiology, University of Oklahoma Health Sciences Center, OK, USA; Center for Geroscience and Healthy Brain Aging, University of Oklahoma Health Sciences Center, OK, USA; Harold Hamm Diabetes Center, University of Oklahoma Health Sciences Center, OK, USA. Electronic address: carlosmanlio-diazgarcia@ouhsc.edu.

PMID: 38430790
PMCID: PMC10990772 (available on 2025-05-01)
DOI: 10.1016/j.ceca.2024.102854

Abstract

The necroptotic effector molecule MLKL accumulates in neurons over the lifespan of mice, and its downregulation has the potential to improve cognition through neuroinflammation, and changes in the abundance of synaptic proteins and enzymes in the central nervous system. Notwithstanding, direct evidence of cell-autonomous effects of MLKL expression on neuronal physiology and metabolism are lacking. Here, we tested whether the overexpression of MLKL in the absence of cell death in the neuronal cell line Neuro-2a recapitulates some of the hallmarks of aging at the cellular level. Using genetically-encoded fluorescent biosensors, we monitored the cytosolic and mitochondrial Ca²⁺ levels, along with the cytosolic concentrations of several metabolites involved in energy metabolism (lactate, glucose, ATP) and oxidative stress (oxidized/reduced glutathione). We found that MLKL overexpression marginally decreased cell viability, however, it led to reduced cytosolic and mitochondrial Ca²⁺ elevations in response to Ca²⁺ influx from the extracellular space. On the contrary, Ca²⁺ signals were elevated after mobilizing Ca²⁺ from the endoplasmic reticulum. Transient elevations in cytosolic Ca²⁺, mimicking neuronal stimulation, lead to higher lactate levels and lower glucose concentrations in Neuro-2a cells when overexpressing MLKL, which suggest enhanced neuronal glycolysis. Despite these alterations, energy levels and glutathione redox state in the cell bodies remained largely preserved after inducing MLKL overexpression for 24-48 h. Taken together, our proof-of-concept experiments are consistent with the hypothesis that MLKL overexpression in the absence of cell death contributes to both Ca²⁺ and metabolic dyshomeostasis, which are cellular hallmarks of brain aging.

Keywords: Aging; Calcium; Lactate; MLKL; Metabolism; Mitochondria; Necroptosis; Neuron.

PubMed Disclaimer

Conflict of interest statement

Declaration of competing interest The authors declare no conflict of interest.

References

1. Akerboom J, Carreras Calderón N, Tian L, Wabnig S, Prigge M, Tolö J, Gordus A, Orger MB, Severi KE, Macklin JJ, Patel R, Pulver SR, Wardill TJ, Fischer E, Schüler C, Chen T-W, Sarkisyan KS, Marvin JS, Bargmann CI, … Looger LL (2013). Genetically encoded calcium indicators for multi-color neural activity imaging and combination with optogenetics. Frontiers in Molecular Neuroscience, 6. 10.3389/fnmol.2013.00002 - DOI - PMC - PubMed
1. Arrázola MS, Lira M, Véliz-Valverde F, Quiroz G, Iqbal S, Eaton SL, Lamont DJ, Huerta H, Ureta G, Bernales S, Cárdenas JC, Cerpa W, Wishart TM, & Court FA (2023). Necroptosis inhibition counteracts neurodegeneration, memory decline, and key hallmarks of aging, promoting brain rejuvenation. Aging Cell, 22(5), e13814. 10.1111/acel.13814 - DOI - PMC - PubMed
1. Arrázola MS, Saquel C, Catalán RJ, Barrientos SA, Hernandez DE, Martínez NW, Catenaccio A, & Court FA (2019). Axonal Degeneration Is Mediated by Necroptosis Activation. The Journal of Neuroscience: The Official Journal of the Society for Neuroscience, 39(20), 3832–3844. 10.1523/JNEUROSCI.0881-18.2019 - DOI - PMC - PubMed
1. Bishop NA, Lu T, & Yankner BA (2010). Neural mechanisms of ageing and cognitive decline. Nature, 464(7288), 529–535. 10.1038/nature08983 - DOI - PMC - PubMed
1. Bröer S, Schneider HP, Bröer A, Rahman B, Hamprecht B, & Deitmer JW (1998). Characterization of the monocarboxylate transporter 1 expressed in Xenopus laevis oocytes by changes in cytosolic pH. Biochemical Journal, 333(Pt 1), 167–174. - PMC - PubMed

MeSH terms

Substances

Grants and funding

P20 GM125528/GM/NIGMS NIH HHS/United States

LinkOut - more resources

Full Text Sources
- ClinicalKey
- Elsevier Science
Miscellaneous
- NCI CPTAC Assay Portal

[1] Akerboom J, Carreras Calderón N, Tian L, Wabnig S, Prigge M, Tolö J, Gordus A, Orger MB, Severi KE, Macklin JJ, Patel R, Pulver SR, Wardill TJ, Fischer E, Schüler C, Chen T-W, Sarkisyan KS, Marvin JS, Bargmann CI, … Looger LL (2013). Genetically encoded calcium indicators for multi-color neural activity imaging and combination with optogenetics. Frontiers in Molecular Neuroscience, 6. 10.3389/fnmol.2013.00002 - DOI - PMC - PubMed

[2] Akerboom J, Carreras Calderón N, Tian L, Wabnig S, Prigge M, Tolö J, Gordus A, Orger MB, Severi KE, Macklin JJ, Patel R, Pulver SR, Wardill TJ, Fischer E, Schüler C, Chen T-W, Sarkisyan KS, Marvin JS, Bargmann CI, … Looger LL (2013). Genetically encoded calcium indicators for multi-color neural activity imaging and combination with optogenetics. Frontiers in Molecular Neuroscience, 6. 10.3389/fnmol.2013.00002 - DOI - PMC - PubMed

[3] Arrázola MS, Lira M, Véliz-Valverde F, Quiroz G, Iqbal S, Eaton SL, Lamont DJ, Huerta H, Ureta G, Bernales S, Cárdenas JC, Cerpa W, Wishart TM, & Court FA (2023). Necroptosis inhibition counteracts neurodegeneration, memory decline, and key hallmarks of aging, promoting brain rejuvenation. Aging Cell, 22(5), e13814. 10.1111/acel.13814 - DOI - PMC - PubMed

[4] Arrázola MS, Lira M, Véliz-Valverde F, Quiroz G, Iqbal S, Eaton SL, Lamont DJ, Huerta H, Ureta G, Bernales S, Cárdenas JC, Cerpa W, Wishart TM, & Court FA (2023). Necroptosis inhibition counteracts neurodegeneration, memory decline, and key hallmarks of aging, promoting brain rejuvenation. Aging Cell, 22(5), e13814. 10.1111/acel.13814 - DOI - PMC - PubMed

[5] Arrázola MS, Saquel C, Catalán RJ, Barrientos SA, Hernandez DE, Martínez NW, Catenaccio A, & Court FA (2019). Axonal Degeneration Is Mediated by Necroptosis Activation. The Journal of Neuroscience: The Official Journal of the Society for Neuroscience, 39(20), 3832–3844. 10.1523/JNEUROSCI.0881-18.2019 - DOI - PMC - PubMed

[6] Arrázola MS, Saquel C, Catalán RJ, Barrientos SA, Hernandez DE, Martínez NW, Catenaccio A, & Court FA (2019). Axonal Degeneration Is Mediated by Necroptosis Activation. The Journal of Neuroscience: The Official Journal of the Society for Neuroscience, 39(20), 3832–3844. 10.1523/JNEUROSCI.0881-18.2019 - DOI - PMC - PubMed

[7] Bishop NA, Lu T, & Yankner BA (2010). Neural mechanisms of ageing and cognitive decline. Nature, 464(7288), 529–535. 10.1038/nature08983 - DOI - PMC - PubMed

[8] Bishop NA, Lu T, & Yankner BA (2010). Neural mechanisms of ageing and cognitive decline. Nature, 464(7288), 529–535. 10.1038/nature08983 - DOI - PMC - PubMed

[9] Bröer S, Schneider HP, Bröer A, Rahman B, Hamprecht B, & Deitmer JW (1998). Characterization of the monocarboxylate transporter 1 expressed in Xenopus laevis oocytes by changes in cytosolic pH. Biochemical Journal, 333(Pt 1), 167–174. - PMC - PubMed

[10] Bröer S, Schneider HP, Bröer A, Rahman B, Hamprecht B, & Deitmer JW (1998). Characterization of the monocarboxylate transporter 1 expressed in Xenopus laevis oocytes by changes in cytosolic pH. Biochemical Journal, 333(Pt 1), 167–174. - PMC - PubMed

Save citation to file

Email citation

Add to Collections

Add to My Bibliography

Your saved search

Create a file for external citation management software

Your RSS Feed

MLKL overexpression leads to Ca²⁺ and metabolic dyshomeostasis in a neuronal cell model

Affiliations

MLKL overexpression leads to Ca²⁺ and metabolic dyshomeostasis in a neuronal cell model

Authors

Affiliations

Abstract

Conflict of interest statement

Similar articles

References

MeSH terms

Substances

Grants and funding

LinkOut - more resources

Full Text Sources

Miscellaneous

Abstract

Conflict of interest statement

Similar articles

References

MeSH terms

Substances

Related information

Grants and funding

LinkOut - more resources

Full Text Sources

Miscellaneous