Identification of Biomarkers Affecting Cryopreservation Recovery Ratio in Ram Spermatozoa Using Tandem Mass Tags (TMT)-Based Quantitative Proteomics Approach
Abstract
Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Experimental Design
2.2. Animals and Ethics
2.3. Sample Collection and Preparation
2.4. Quantitative Mass Spectrometry Analysis
2.4.1. Protein Extraction and Digestion (PTM)
2.4.2. TMT Labeling
2.4.3. High-Performance Liquid Chromatography (HPLC) Fractionation and LC-MS/MS Analysis
2.4.4. Database Search and Bioinformatic Analysis
2.5. PRM Validation
2.6. Statistical Analysis
3. Results
3.1. Comparison of Sample Parameters between HCR and LCR
3.2. Proteomic Profile of Spermatozoa
3.3. Validation of the Selected DEPs by PRM
3.4. Bioinformatics Analysis of DEPs
3.5. Protein-Protein Interaction Analysis
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Group | Concentration (Million/mL) | Age (Months) | Weight (kg) | Breed |
---|---|---|---|---|
HCR | 3861.60 ± 296.11 | 37.00 ± 0.58 | 98.33 ± 0.88 | Dorper sheep |
LCR | 4191.53 ± 314.09 | 36.33 ± 0.88 | 96.33 ± 0.88 | Dorper sheep |
Parameters | HCR | LCR | p-Value |
---|---|---|---|
Fresh sperm | |||
MOT (%) | 82.3 ± 0.006 | 80.9 ± 0.02 | 0.41 |
VCL (μm/s) | 101.3 ± 2.98 | 103.1 ± 3.10 | 0.70 |
VSL (μm/s) | 43 ± 2.65 | 38.725 ± 2.14 | 0.25 |
VAP (μm/s) | 60.8 ± 1.33 | 58.1 ± 1.43 | 0.18 |
LIN (%) | 42.8 ± 2.71 | 37.9 ± 2.34 | 0.21 |
STR (%) | 63.8 ± 2.17 | 60.6 ± 1.84 | 0.31 |
ALH (μm) | 3.6 ± 0.16 | 3.8 ± 0.18 | 0.33 |
BCF (Hz) | 6.2 ± 0.15 | 6.1 ± 0.20 | 0.60 |
Cryo-recovered sperm | |||
MOT (%) | 56.6 ± 0.02 *** | 22.1 ± 0.02 *** | <0.001 |
VCL (μm/s) | 80.2 ± 1.94 *** | 48.5 ± 2.49 *** | <0.001 |
VSL (μm/s) | 31.5 ± 1.27 *** | 22.2 ± 1.33 *** | <0.001 |
VAP (μm/s) | 45.4 ± 1.50 *** | 36.9 ± 2.06 *** | <0.001 |
LIN (%) | 39.7 ± 1.95 * | 45.7 ± 1.72 * | <0.05 |
STR (%) | 61.8 ± 1.38 * | 67.1 ± 1.42 * | <0.05 |
ALH (μm) | 3.3 ± 0.12 *** | 2.6 ± 0.09 *** | <0.001 |
BCF (Hz) | 5.1 ± 0.11 *** | 6.9 ± 0.25 *** | <0.001 |
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Ren, C.; Sun, Z.; Chen, Y.; Chen, J.; Wang, S.; Liu, Q.; Wang, P.; Cheng, X.; Zhang, Z.; Wang, Q. Identification of Biomarkers Affecting Cryopreservation Recovery Ratio in Ram Spermatozoa Using Tandem Mass Tags (TMT)-Based Quantitative Proteomics Approach. Animals 2023, 13, 2368. https://doi.org/10.3390/ani13142368
Ren C, Sun Z, Chen Y, Chen J, Wang S, Liu Q, Wang P, Cheng X, Zhang Z, Wang Q. Identification of Biomarkers Affecting Cryopreservation Recovery Ratio in Ram Spermatozoa Using Tandem Mass Tags (TMT)-Based Quantitative Proteomics Approach. Animals. 2023; 13(14):2368. https://doi.org/10.3390/ani13142368
Chicago/Turabian StyleRen, Chunhuan, Zhipeng Sun, Yale Chen, Jiahong Chen, Shijia Wang, Qingqing Liu, Penghui Wang, Xiao Cheng, Zijun Zhang, and Qiangjun Wang. 2023. "Identification of Biomarkers Affecting Cryopreservation Recovery Ratio in Ram Spermatozoa Using Tandem Mass Tags (TMT)-Based Quantitative Proteomics Approach" Animals 13, no. 14: 2368. https://doi.org/10.3390/ani13142368
APA StyleRen, C., Sun, Z., Chen, Y., Chen, J., Wang, S., Liu, Q., Wang, P., Cheng, X., Zhang, Z., & Wang, Q. (2023). Identification of Biomarkers Affecting Cryopreservation Recovery Ratio in Ram Spermatozoa Using Tandem Mass Tags (TMT)-Based Quantitative Proteomics Approach. Animals, 13(14), 2368. https://doi.org/10.3390/ani13142368