Sperm and Spermatids Contain Different Proteins and Bind Distinct Egg Factors
Abstract
:1. Introduction
2. Results and Discussion
2.1. Sperm and Spermatids Differ in Their Nuclear Protein Composition
2.1.1. Sperm and Spermatids Contain Different Proteins
2.1.2. Sperm-Specific Proteins Contain Several Factors Which May Support Subsequent Embryonic Development
2.1.3. Chromatin and Nuclear Proteins Are Enriched in Sperm
2.2. Sperm and Spermatids Attract Distinct Egg Factors
2.2.1. Dramatic Changes in Protein Composition after the Egg Extract Treatment
2.2.2. Many Egg Proteins Bind Differentially to Sperm and to Spermatid Chromatin
2.2.3. Known Chromatin Remodelling Factors and Replication Proteins Were Incorporated from Egg Extracts into both Sperm and Spermatids
2.2.4. Distinct Protein Isoforms Identified in Extract-Treated Sperm- and Spermatids
2.2.5. Many Egg-Derived Chromatin Factors Bind Preferentially to Sperm Nuclei
2.2.6. GO (Gene Ontology) Analysis Confirms the Overrepresentation of Egg Chromatin Remodelling Factors Binding Specifically to Sperm Nuclei
2.3. Validation of Mass Spectrometry Results by Immunoblotting
3. Experimental Section
3.1. Isolation and Permeabilization of Sperm and Spermatids
3.2. Isolation of Proteins and Sample Preparation for 2D-DIGE
3.3. 2D-DIGE Analysis
3.4. Spot Excision and Protein Preparation for Mass Spectrometry Analysis
3.5. LC-MS/MS Analysis and Protein Identification
3.6. Interphase Egg Extract Preparation
3.7. Egg Extract Treatment and Protein Isolation for Mass Spectrometry Analysis of Egg Proteins Incorporated into Sperm or Spermatid Chromatin
3.8. Immunoblotting Analysis
4. Conclusions
Supplementary Materials
Supplementary Files
Supplementary File 1Acknowledgments
Author Contributions
Conflicts of Interest
References
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Teperek, M.; Miyamoto, K.; Simeone, A.; Feret, R.; Deery, M.J.; Gurdon, J.B.; Jullien, J. Sperm and Spermatids Contain Different Proteins and Bind Distinct Egg Factors. Int. J. Mol. Sci. 2014, 15, 16719-16740. https://doi.org/10.3390/ijms150916719
Teperek M, Miyamoto K, Simeone A, Feret R, Deery MJ, Gurdon JB, Jullien J. Sperm and Spermatids Contain Different Proteins and Bind Distinct Egg Factors. International Journal of Molecular Sciences. 2014; 15(9):16719-16740. https://doi.org/10.3390/ijms150916719
Chicago/Turabian StyleTeperek, Marta, Kei Miyamoto, Angela Simeone, Renata Feret, Michael J. Deery, John B. Gurdon, and Jerome Jullien. 2014. "Sperm and Spermatids Contain Different Proteins and Bind Distinct Egg Factors" International Journal of Molecular Sciences 15, no. 9: 16719-16740. https://doi.org/10.3390/ijms150916719