TGF-β Signaling Regulates SLC8A3 Expression and Prevents Oxidative Stress in Developing Midbrain Dopaminergic and Dorsal Raphe Serotonergic Neurons
Abstract
:1. Introduction
2. Results
2.1. SLC8A3 Expression is Regulated by TGF-β Signaling
2.2. Anti-Apoptotic Action of SLC8A3
2.3. Cell-Type-Dependent Regulation of SLC8A3 by TGF-β
2.4. TGF-β Canonical Pathway and p38 Signaling Pathway Regulate Basal SLC8A3 Protein Expression
2.5. Smad4 Binds to Slc8a3 Promoter
2.6. Knockdown of Slc8a3 is Associated with Oxidative Stress
2.7. Cell-Type-Specific Deletion of TGF-β Signaling is Associated with Oxidative Stress
3. Discussion
4. Materials and Methods
4.1. Animals
4.2. Genotyping
4.3. Immunohistochemistry
4.4. Cell Culture of E14 Ventral Mesencephalon and Ventral Hindbrain
4.5. Cell Culture of MN9D and LUHMES Cells
4.6. Immunocytochemistry
4.7. Image Acquisition and Analysis
4.8. Immunoblotting
4.9. Chromatin Immunoprecipitation Assay
4.10. Transient Transfection of MN9D Cells
4.11. Statistics
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
4-HNE | 4-hydroxynonenal |
5-HT | 5-hydroxytryptamine |
Bcl-xL | B-cell lymphoma-extra large |
BDNF | Brain derived neurotrophic factor |
cKO | Conditional knockout |
DMEM | Dulbecco’s Modified Eagle Medium |
DR | Dorsal raphe |
E# | Embryonic day |
En1 | Engrailed 1 |
ERK1/2 | Extracellular signal–regulated kinases-1/2 |
GAPDH | Glyceraldehyde 3-phosphate dehydrogenase |
H3 | Histone 3 |
JNK1/2 | c-Jun N-terminal kinases |
L-type Ca2+ | Long-lasting calcium channel |
LUHMES | Lund Human Mesencephalic |
MAPK | Mitogen-activated protein kinase |
MEK | Mitogen-activated protein kinase kinase |
mDA | Midbrain dopaminergic |
MDA | Malondialdehyde |
NCX3 | Na+/Ca2+ exchanger 3 |
NGF | Nerve growth factor |
PC12 | Pheochromocytoma of the rat adrenal medulla |
PD | Parkinson’s disease |
PINK1 | PTEN-induced kinase 1 or PTEN-induced putative kinase 1 |
ROS | Reactive oxygen species |
siRNA | Silencing RNA |
SLC8A3 | Solute carrier Family 8 member A3 |
Smad | Small mother against decapentaplegic |
SNc | Substantia nigra pars compacta |
TGF-β | Transforming growth factor beta |
TRPC | Transient receptor potential canonical channels |
TβRII | Type II TGF-β receptor |
vH | Ventral hindbrain |
VTA | Ventral tegmental area |
Wt | Wildtype |
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Chleilat, E.; Pethe, A.; Pfeifer, D.; Krieglstein, K.; Roussa, E. TGF-β Signaling Regulates SLC8A3 Expression and Prevents Oxidative Stress in Developing Midbrain Dopaminergic and Dorsal Raphe Serotonergic Neurons. Int. J. Mol. Sci. 2020, 21, 2735. https://doi.org/10.3390/ijms21082735
Chleilat E, Pethe A, Pfeifer D, Krieglstein K, Roussa E. TGF-β Signaling Regulates SLC8A3 Expression and Prevents Oxidative Stress in Developing Midbrain Dopaminergic and Dorsal Raphe Serotonergic Neurons. International Journal of Molecular Sciences. 2020; 21(8):2735. https://doi.org/10.3390/ijms21082735
Chicago/Turabian StyleChleilat, Enaam, Abhishek Pethe, Dietmar Pfeifer, Kerstin Krieglstein, and Eleni Roussa. 2020. "TGF-β Signaling Regulates SLC8A3 Expression and Prevents Oxidative Stress in Developing Midbrain Dopaminergic and Dorsal Raphe Serotonergic Neurons" International Journal of Molecular Sciences 21, no. 8: 2735. https://doi.org/10.3390/ijms21082735