Abstract
Proteomics is a technology that allows to decipher the molecular networks involved in the regulation of biological processes such as spermatogenesis. Sertoli cells (SCs) are key players in the paracrine control of this process. Envisioning to increase the knowledge on the molecular networks harbored in SCs, we propose a methodology based on GeLC-MS/MS for the characterization of these cells’ proteome. Proteins are separated by SDS-PAGE hyphenated to HPLC and identified by mass spectrometry. The integration of data with bioinformatics tools such as ClueGO + CluePedia from Cytoscape allows the identification of the biological pathways more prevalent in SCs, and that might be modulated by pathophysiological conditions. Moreover, the proteome analysis with tools as SignalP/SecretomeP highlights the proteins more prone to be secreted and involved in the paracrine control of germ cells, which might also be deregulated by diseases.
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Acknowledgments
The authors thank Portuguese Foundation for Science and Technology (FCT), European Union, QREN, FEDER, and COMPETE for funding the iBiMED (UID/BIM/04501/2013), QOPNA (UID/QUI/00062/2013), UnIC (UID/IC/00051/2013), research project (POCI-01-0145-FEDER-016728; PTDC/DTP-DES/6077/2014), and RV’s (IF/00286/2015) and FT’s (SFRH/BD/111633/2015) fellowship grants.
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1 Electronic Supplementary Materials
Supplementary Table S1
List of proteins previously identified in mouse SCs [5] (XLSX 29 kb)
Supplementary Table S2
List of proteins retrieved from SecretomeP and SignalP analysis (XLSX 635 kb)
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Ferreira, R., Trindade, F., Vitorino, R. (2018). Proteome Profiling of Sertoli Cells Using a GeLC-MS/MS Strategy. In: Alves, M., Oliveira, P. (eds) Sertoli Cells. Methods in Molecular Biology, vol 1748. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7698-0_13
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DOI: https://doi.org/10.1007/978-1-4939-7698-0_13
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