Transcriptomic Analysis in Diabetic Nephropathy of Streptozotocin-Induced Diabetic Rats
Abstract
:1. Introduction
2. Results and Discussion
2.1. Biochemical Parameters in Healthy and Diabetic Rats
2.2. Kidney Histopathology
2.3. Differential Expression of Genes
2.4. Analysis of Sequence Homology of EST’s
2.5. Differential Expression of Selected Transcripts from the Subtractive Library
2.6. Related Function of Transcript Selected from the Subtractive Library
2.6.1. Metabolism Protein
2.6.2. Transporter and Channels
2.6.3. Transcription Factors
2.6.4. Unknown Genes
3. Experimental Section
3.1. Induction of Diabetes and Experimental Protocol
3.2. Assays of Serum, Urine and Kidney Tissue Samples
3.3. Kidney Histopathology
3.4. RNA Isolation
3.5. PCR-Selected cDNA Subtraction
3.6. Cloning, Screening, and cDNA Sequencing Subtracted
3.7. Northern Blot Analysis
3.8. Statistical Analysis of Expression Data
4. Conclusions
Acknowledgments
- Conflict of InterestThe authors declare no conflict of interest.
References
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Parameters | Healthy control | Diabetic control |
---|---|---|
Physiological: | ||
Body weight (g) | 379.7 ± 12.2 a | 248.6 ± 8.3 b |
Kidney weight (g/100 g BW) | 0.60 ± 0.01 b | 0.90 ± 0.02 a |
Food intake (g/rat/day) | 28.5 ± 0.6 b | 56.7 ± 1.9 a |
Water intake (mL/rat/day) | 40.6 ± 3.6 b | 385.8 ± 9.6 a |
Urine output (mL/rat/day) | 22.4 ± 2.9 b | 43.6 ± 3.2 a |
Creatinine clearance (mL/min) | 1.63 ± 0.04 a | 0.78 ± 0.03 b |
Urine (mg/24h): | ||
Albumin | 1.3 ± 0.2 b | 2.6 ± 0.4 a |
Protein | 4.3 ± 0.3 b | 7.2 ± 1.2 a |
Urea | 5.1 ± 0.6 b | 30.4 ± 3.2 a |
Serum: | ||
Fasting glucose (mg/dL) | 52.0 ± 2.0 b | 356 ± 29.1 a |
Insulin (μU/mL) | 19.1 ± 3.4 a | 9.9 ± 1.0 b |
Urea (mg/dL) | 23.7 ± 1.8 b | 80.1 ± 4.9 a |
Function | Accession No. | Blast result | pb | Fold change |
---|---|---|---|---|
(a) Metabolism proteins | NM_172034 | Farnesyltransferase beta subunit (Fntb) | 281 | 0.742 |
AA874879.1 | NADH dehydrogenase | 388 | 0.623 | |
NM_017034 | Threonine-protein kinase (PIM-1) | 675 | 0.502 | |
NM_021577 | Argininosuccinate lyase (ASL) | 626 | 0.517 | |
NM_053774.2 | Ubiquitin-specific peptidase 2 (USP2) | 251 | 0.676 | |
NM_022243.1 | Hydroxyisobutyrate dehydrogenase (Hibadh) | 368 | 0.954 | |
NM_022933.2 | Helicase DNA binding protein 8 (Chd8) | 202 | 0.922 | |
NM_031795.2 | UDP glucose ceramide glycosyltransferase (Ugcg) | 346 | 0.564 | |
AF227741.1 | Serine/threonine-protein kinase (WNK1) | 423 | 0.870 | |
NM_024160.1 | Cytochrome b-245, alpha polypeptide (Cyba) | 663 | 0.792 | |
NM_001009668.1 | Electron-transfer-flavoprotein, alpha polypeptide (Etfa) | 373 | 0.506 | |
M29579.1 | Glutamine-synthetase (GS) | 372 | 0.640 | |
XM_221132.5 | Ubiquitin-conjugating enzyme E2O (Ube2o) | 226 | 0.858 | |
(b) Transporters and channels | NM_052983.2 | Solute carrier family 5 (sodium iodide symporter), member 5 (Slc5a5) | 280 | 0.906 |
NM_016996.1 | Calcium-sensing receptor (Casr) | 417 | 0.413 | |
D50497.1 | CIC-type chloride channel (ClC-5) | 242 | 0.786 | |
(c) Transcription factors | NM_001191711 | DEAD box polypeptide 20 (Ddx20) | 519 | 0.793 |
NM_013141.2 | Peroxisome proliferator-activated receptor delta (Ppard) | 580 | 0.631 |
Function | Accession No. | Blast result | pb | Fold change |
---|---|---|---|---|
(a) Metabolism proteins | NM_001009637.1 | Leucyl-tRNA synthetase (Lars) | 511 | 1.011 |
NM_001039346.1 | Zinc finger, DHHC-type containing 16 (Zdhhc16) | 250 | 1.360 | |
NM_001159739.1 | Glutathione-S-transferase (Gsta5) | 237 | 1.133 | |
NM_032080.1 | Glycogen synthase kinase 3 beta (Gsk3b) | 302 | 1.143 | |
NM_001007620.1 | Pyruvate dehydrogenase beta (Pdhb) | 200 | 1.697 | |
NM_134417.1 | Inositol polyphosphate multikinase (Ipmk) | 295 | 1.473 | |
NM_001004214.1 | NAD(P)H dehydrogenase, quinone 2 (Nqo2) | 596 | 1.010 | |
NM_ NP_446292.2 | Gamma-glutamyltranspeptidase 1 (Ggtp1) | 636 | 1.755 | |
NM_001007235.1 | Inositol 1,4,5-triphosphate receptor, type 1 (Itpr1) | 504 | 1.093 | |
NM_012570.1 | Glutamate dehydrogenase 1 (Glud1) | 327 | 1.103 | |
NM_001004252.1 | Phenylalanyl-tRNA synthetase, beta subunit (Farsb) | 301 | 1.012 | |
NM_017072.1 | Carbamoyl-phosphate synthase subunit 1 (Cps1) | 591 | 1.38 | |
(b) Transporters and channels | NM_173338.1 | Solute carrier organic anion transporter family (SIco6c1) | 414 | 1.777 |
NM_134468.1 | Calcium-and calmodulindependent protein kinase type 1 (Camk1) | 256 | 1.226 | |
NM_012517.2 | Calcium channel, voltage-dependent, L type, alpha 1C subunit (Cacna1c) | 200 | 1.379 | |
(c) Transcription factors | NM_001127373.1 | ADNP homeobox 2 (Adnp2) | 380 | 1.967 |
NM_031346.1 | ROD1 regulator of differentiation 1 (S. pombe)(Rod1) | 204 | 1.963 | |
NM_001033694.1 | Sterol regulatory element binding transcription factor 2 (Srebf2) | 766 | 1.331 |
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Lomas-Soria, C.; Ramos-Gómez, M.; Guevara-Olvera, L.; Guevara-González, R.; Torres-Pacheco, I.; Gallegos-Corona, M.A.; Reynoso-Camacho, R. Transcriptomic Analysis in Diabetic Nephropathy of Streptozotocin-Induced Diabetic Rats. Int. J. Mol. Sci. 2011, 12, 8431-8448. https://doi.org/10.3390/ijms12128431
Lomas-Soria C, Ramos-Gómez M, Guevara-Olvera L, Guevara-González R, Torres-Pacheco I, Gallegos-Corona MA, Reynoso-Camacho R. Transcriptomic Analysis in Diabetic Nephropathy of Streptozotocin-Induced Diabetic Rats. International Journal of Molecular Sciences. 2011; 12(12):8431-8448. https://doi.org/10.3390/ijms12128431
Chicago/Turabian StyleLomas-Soria, Consuelo, Minerva Ramos-Gómez, Lorenzo Guevara-Olvera, Ramón Guevara-González, Irineo Torres-Pacheco, Marco A. Gallegos-Corona, and Rosalía Reynoso-Camacho. 2011. "Transcriptomic Analysis in Diabetic Nephropathy of Streptozotocin-Induced Diabetic Rats" International Journal of Molecular Sciences 12, no. 12: 8431-8448. https://doi.org/10.3390/ijms12128431