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Proteome Profiling of Sertoli Cells Using a GeLC-MS/MS Strategy

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Book cover Sertoli Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1748))

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.

Author information

Authors and Affiliations

  1. QOPNA, Department of Chemistry, University of Aveiro, Aveiro, Portugal

    Rita Ferreira

  2. iBiMED, Department of Medical Sciences, University of Aveiro, Aveiro, Portugal

    Fábio Trindade & Rui Vitorino

  3. UnIC, Departamento de Cirurgia e Fisiologia, Faculdade de Medicina, Universidade do Porto, Porto, Portugal

    Fábio Trindade & Rui Vitorino

Authors
  1. Rita Ferreira
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  2. Fábio Trindade
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  3. Rui Vitorino
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Corresponding author

Correspondence to Rui Vitorino .

Editor information

Editors and Affiliations

  1. Department of Microscopy, Laboratory of Cell Biology and Unit for Multidisciplinary Research in Biomedicine (UMIB), Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto, Porto, Portugal

    Marco G. Alves

  2. Department of Microscopy, Laboratory of Cell Biology and Unit for Multidisciplinary Research in Biomedicine (UMIB), Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto, Porto, Portugal

    Pedro F. Oliveira

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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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7697-3

  • Online ISBN: 978-1-4939-7698-0

  • eBook Packages: Springer Protocols

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