Mass Spectrometry Analysis of Neurotransmitter Shifting during Neurogenesis and Neurodegeneration of PC12 Cells
Abstract
:1. Introduction
2. Results
2.1. Effects of Rotenone on the Morphology of Cells That Underwent Neurodifferentiation and Degeneration
2.2. Neurotransmitter Analysis
2.3. Activation of Dopaminergic Metabolism through PC12 Differentiation
2.4. Effects of Rotenone Pretreatment on PC12 Differentiation
2.5. Effects of Rotenone Pretreatment on Neurotransmitter Release
2.6. Effects of Rotenone on Neurotransmitters during Neurodegeneration
3. Discussion
3.1. Activation of Dopaminergic Metabolites during Neuron Differentiation
3.2. Rotenone Treatment Shifts Dopamine Metabolism and Activates Serotoninergic Pathway
3.3. Detrimental Effect of Rotenone on Differentiated PC12 Cells
4. Materials and Methods
4.1. Materials
4.2. PC12 Cell Culture and Differentiation
4.3. Inhibition of Cell Differentiation by Rotenone
4.4. Effects of Rotenone on Neurodegeneration
4.5. Extraction of Neurotransmitters in Cells and Medium
4.6. Liquid Chromatography–Mass Spectrometry Analysis
4.7. Quantification and Semiquantification of Neurotransmitters
4.8. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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No. | Observed Mass m/z | Identified Mass m/z | Neurotransmitter | Retention Time (min) | Section (min) | Collision (eV) |
---|---|---|---|---|---|---|
1 | 104.07 | 87 | γ-Aminobutyric acid | 4.0–4.8 | I (0–14) | 30 |
2 | 112.08 | 95 | Histamine | 3.7–4.8 | I (0–14) | 30 |
3 | 146.11 | 87 | Acetylcholine | 5.9–9.0 | I (0–14) | 20 |
4 | 148.06 | 84 | Glutamate | 4.7–5.9 | I (0–14) | 25 |
5 | 153.09 | 88 | Glutamate-D5 | 4.7–5.9 | I (0–14) | 25 |
6 | 154.08 | 91 | Dopamine | 9.4–12.0 | I (0–14) | 25 |
7 | 158.11 | 95 | Dopamine-D4 | 9.4–12.0 | I (0–14) | 25 |
8 | 170.08 | 152 | Norepinephrine | 4.8–5.6 | I (0–14) | 25 |
9 | 176.11 | 158 | Norepinephrine-D6 | 4.8–5.6 | I (0–14) | 25 |
10 | 184.09 | 166 | Epinephrine | 5.9–8.0 | I (0–14) | 25 |
11 | 138.05 | 121 | Tyramine | 19.7–20.1 | II (15–25) | 30 |
12 | 166.08 | 120 | Phenylalanine | 19.4–20.3 | II (15–25) | 25 |
13 | 168.10 | 91 | 3-Methoxytyramine | 17.7–18.7 | II (15–25) | 30 |
14 | 177.09 | 160 | Serotonin | 18.1–19.0 | II (15–25) | 25 |
15 | 182.08 | 91 | Tyrosine | 16.1–17.0 | II (15–25) | 30 |
16 | 192.05 | 146 | 5-Hydroxyindoleacetic acid | 21.3–21.9 | II (15–25) | 30 |
17 | 198.13 | 152 | L-dopa | 12.6–15.0 | II (15–25) | 30 |
18 | 205.08 | 188 | Tryptophan | 20.6–21.0 | II (15–25) | 30 |
19 | 233.12 | 174 | Melatonin | 22.1–23.5 | II (15–25) | 30 |
20 | 268.09 | 136 | Adenosine | 16.5–17.3 | II (15–25) | 20 |
21 | 269.10 | 137 | Adenosine-D1 | 16.5–17.3 | II (15–25) | 20 |
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Jao, Y.-N.; Chao, Y.-J.; Chan, J.-F.; Hsu, Y.-H.H. Mass Spectrometry Analysis of Neurotransmitter Shifting during Neurogenesis and Neurodegeneration of PC12 Cells. Int. J. Mol. Sci. 2024, 25, 10399. https://doi.org/10.3390/ijms251910399
Jao Y-N, Chao Y-J, Chan J-F, Hsu Y-HH. Mass Spectrometry Analysis of Neurotransmitter Shifting during Neurogenesis and Neurodegeneration of PC12 Cells. International Journal of Molecular Sciences. 2024; 25(19):10399. https://doi.org/10.3390/ijms251910399
Chicago/Turabian StyleJao, Yu-Ning, Yu-Jen Chao, Jui-Fen Chan, and Yuan-Hao Howard Hsu. 2024. "Mass Spectrometry Analysis of Neurotransmitter Shifting during Neurogenesis and Neurodegeneration of PC12 Cells" International Journal of Molecular Sciences 25, no. 19: 10399. https://doi.org/10.3390/ijms251910399