Effects of Pyridoxine Deficiency on Hippocampal Function and Its Possible Association with V-Type Proton ATPase Subunit B2 and Heat Shock Cognate Protein 70
Abstract
:1. Introduction
2. Materials and Methods
2.1. Experimental Animals
2.2. Novel Object Recognition Test
2.3. High-Performance Liquid Chromatography Analysis
2.4. Immunohistochemistry
2.5. Proteomic Analysis
2.5.1. Protein Preparation for 2DE
2.5.2. Analysis of 2DE Gels
2.5.3. Trypsin Digestion
2.5.4. Protein Identification Using MALDI-TOF MS
2.5.5. Data Searches for Protein Identification
2.6. Validation by Western Blot Analyses
2.7. Data Analysis
2.8. Statistical Analysis
3. Results
3.1. Effects of Pyridoxine Deficiency on PLP Levels in Serum and Brain Tissues
3.2. Effects of Pyridoxine Deficiency on Novel Object Recognition
3.3. Effects of Pyridoxine Deficiency on Monoamine Levels in the Hippocampus
3.4. Effects of Pyridoxine Deficiency on Ki67 and DCX Immunoreactivity
3.5. Protein Identification by 2D-DIGE Followed by MALDI-TOF MS in the Hippocampus
3.6. Validation of Identified Proteins by Western Blot
4. Discussion
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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Proteins (gI Accession Number) | Number of Peptides Matched in the Identified Protein | Protein Sequence Coverage (%) | pI, Mr (kDa) | MOWSE | Fold Decrease |
---|---|---|---|---|---|
APG-1 (705391) | 1 | 1 | 5.53, 95.3 | 39 | |
Zinc finger protein 831 (150010581) | 1 | 0 | 8.18 176.1 | 40 | 2.3 |
Murine valosin-containing protein (55217) | 3 | 3 | 5.14 89.9 | 105 | 2.5 |
Heat-shock protein hsp84 (194027) | 2 | 3 | 4.95 83.5 | 89 | 2.0 |
V-type proton ATPase subunit B2 (17105370) | 38 | 25 | 5.57 56.9 | 638 | 2.3 |
Vacuolar adenosine triphosphatase subunit B (1184661) | 1 | 2 | 5.57 56.9 | 45 | 3.1 |
Acyltransferase (1129118) | 1 | 2 | 8.88 53.5 | 48 | 2.8 |
Anti-DNA immunoglobulin light chain IgG (1870366) | 1 | 8 | 9.11 11.1 | 39 | 2.1 |
Poly(rC)-binding protein 1 (6754994) | 1 | 2 | 6.66 38.0 | 40 | 2.2 |
Guanine nucleotide-binding protein G(I)/G(S)/G(T) subunit beta-1 (6680045) | 12 | 12 | 5.60 38.2 | 243 | 8.8 |
Glyceraldehyde-3-phosphate dehydrogenase (2494630) | 1 | 1 | 8.14 48.4 | 34 | 3.0 |
Charged multivesicular body protein 4b (2807749) | 2 | 11 | 4.76 24.9 | 85 | 2.1 |
Voltage-dependent anion-selective channel protein 2 (6755965) | 3 | 7 | 7.44 32.3 | 102 | 3.4 |
Serine-threonine kinase receptor-associated protein (4063383) | 1 | 3 | 4.99 38.8 | 36 | 2.8 |
Osmotic stress protein 94 (1098541) | 5 | 5 | 5.50 95.2 | 215 | 3.3 |
Proteins (gI Accession Number) | Number of Peptides Matched in the Identified Protein | Protein Sequence Coverage (%) | pI, Mr (kDa) | MOWSE | Fold Increase |
---|---|---|---|---|---|
NADH dehydrogenase (13879366) | 2 | 2 | 5.51 80.7 | 68 | 2.0 |
Heat shock cognate protein 70 (309319) | 49 | 29 | 5.37 71.0 | 1066 | 2.4 |
Heat shock-related 70 kDa protein 2 (31560686) | 20 | 21 | 5.51 69.9 | 620 | 3.5 |
Dihydrolipoamide dehydrogenase (2078522) | 1 | 2 | 7.97 54.7 | 41 | 2.3 |
ATP synthase subunit alpha, mitochondrial precursor (6680748) | 10 | 9 | 9.22 59.8 | 308 | 2.7 |
Aldehyde dehydrogenase, mitochondrial isoform 1 precursor (6753036) | 2 | 3 | 7.53 57.0 | 96 | 2.0 |
Type II keratin subunit protein (4159806) | 1 | 1 | 8.97 65.7 | 40 | 2.8 |
Spermatogenesis-associated protein 7 homolog isoform 1 | 1 | 1 | 6.23 66.2 | 34 | 3.0 |
ATP-specific succinyl-CoA synthetase beta subunit | 16 | 13 | 5.65 46.6 | 366 | 3.6 |
Protein phosphatase type 2A catalytic subunit alpha isoform (3342500) | 1 | 2 | 5.30 36.2 | 48 | 2.4 |
Histidine triad nucleotide-binding protein 1 (33468857) | 1 | 11 | 6.36 13.9 | 41 | 2.1 |
Epidermal keratin subunit I, partial (387397) | 17 | 8 | 5.01 58.0 | 304 | 4.0 |
2-Oxoglutarate dehydrogenase-like, mitochondrial (568987556) | 2 | 83.2 | 96 | 2.4 |
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Jung, H.Y.; Kim, W.; Hahn, K.R.; Kwon, H.J.; Nam, S.M.; Chung, J.Y.; Yoon, Y.S.; Kim, D.W.; Yoo, D.Y.; Hwang, I.K. Effects of Pyridoxine Deficiency on Hippocampal Function and Its Possible Association with V-Type Proton ATPase Subunit B2 and Heat Shock Cognate Protein 70. Cells 2020, 9, 1067. https://doi.org/10.3390/cells9051067
Jung HY, Kim W, Hahn KR, Kwon HJ, Nam SM, Chung JY, Yoon YS, Kim DW, Yoo DY, Hwang IK. Effects of Pyridoxine Deficiency on Hippocampal Function and Its Possible Association with V-Type Proton ATPase Subunit B2 and Heat Shock Cognate Protein 70. Cells. 2020; 9(5):1067. https://doi.org/10.3390/cells9051067
Chicago/Turabian StyleJung, Hyo Young, Woosuk Kim, Kyu Ri Hahn, Hyun Jung Kwon, Sung Min Nam, Jin Young Chung, Yeo Sung Yoon, Dae Won Kim, Dae Young Yoo, and In Koo Hwang. 2020. "Effects of Pyridoxine Deficiency on Hippocampal Function and Its Possible Association with V-Type Proton ATPase Subunit B2 and Heat Shock Cognate Protein 70" Cells 9, no. 5: 1067. https://doi.org/10.3390/cells9051067