Spermatogenesis-associated proteins at different developmental stages of buffalo testicular seminiferous tubules identified by comparative proteomic analysis

Proteomics. 2016 Jul;16(14):2005-18. doi: 10.1002/pmic.201500547. Epub 2016 Jun 27.

Abstract

The testicular seminiferous tubules contain Sertoli cells and different types of spermatogenic cells. They provide the microenvironment for spermatogenesis, but the precise molecular mechanism of spermatogenesis is still not well known. Here, we have employed tandem mass tag coupled to LC-MS/MS with the high-throughput quantitative proteomics technology to explore the protein expression from buffalo testicular seminiferous tubules at three different developmental stages (prepuberty, puberty, and postpuberty). The results show 304 differentially expressed proteins with a ≥2-fold change, and bioinformatics analysis indicates that 27 of these may be associated with spermatogenesis. Expression patterns of seven selected proteins were verified via Western blot and quantitative RT-PCR analysis, and further cellular localizations of these proteins by immunohistochemical or immunofluorescence analysis. Taken together, the results provide potential molecular markers of spermatogenesis and provide a rich resource for further studies on male reproduction regulation.

Keywords: Animal proteomics; Buffalo; Spermatogenesis-associated proteins; Tandem mass tag; Testicular seminiferous tubules.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • A Kinase Anchor Proteins / genetics
  • A Kinase Anchor Proteins / metabolism
  • Animals
  • Buffaloes
  • Chromatography, Liquid
  • Gene Expression Regulation, Developmental*
  • Gene Ontology
  • Heat-Shock Proteins / genetics
  • Heat-Shock Proteins / metabolism
  • Male
  • Molecular Sequence Annotation
  • Proteome / genetics*
  • Proteome / metabolism
  • Proteomics / methods
  • Seminiferous Tubules / cytology
  • Seminiferous Tubules / growth & development
  • Seminiferous Tubules / metabolism*
  • Sertoli Cells / cytology
  • Sertoli Cells / metabolism*
  • Sexual Maturation / genetics
  • Spermatogenesis / genetics*
  • Tandem Mass Spectrometry
  • rho GTP-Binding Proteins / genetics
  • rho GTP-Binding Proteins / metabolism

Substances

  • A Kinase Anchor Proteins
  • Heat-Shock Proteins
  • Proteome
  • rho GTP-Binding Proteins