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An integrative omics strategy to assess the germ cell secretome and to decipher sertoli-germ cell crosstalk in the Mammalian testis.

Chalmel F, Com E, Lavigne R, Hernio N, Teixeira-Gomes AP, Dacheux JL, Pineau C - PLoS ONE (2014)

Bottom Line: Two interactions, APOH/CDC42 and APP/NGFR, were validated in situ, in a proximity ligation assay (PLA).Our findings also demonstrate that this "integrative omics" strategy is powerful enough for data mining and highlighting meaningful cell-cell communication events between different types of cells in a complex tissue, via a biological fluid.This integrative strategy could be applied more widely, to gain access to secretomes that have proved difficult to study whilst avoiding the limitations of in vitro culture.

View Article: PubMed Central - PubMed

Affiliation: IRSET, Inserm U1085, Campus de Beaulieu, Rennes, France.

ABSTRACT
Mammalian spermatogenesis, which takes place in complex testicular structures called seminiferous tubules, is a highly specialized process controlled by the integration of juxtacrine, paracrine and endocrine information. Within the seminiferous tubules, the germ cells and Sertoli cells are surrounded by testicular fluid (TF), which probably contains most of the secreted proteins involved in crosstalk between these cells. It has already been established that germ cells can modulate somatic Sertoli cell function through the secretion of diffusible factors. We studied the germ cell secretome, which was previously considered inaccessible, by analyzing the TF collected by microsurgery in an "integrative omics" strategy combining proteomics, transcriptomics, genomics and interactomics data. This approach identified a set of proteins preferentially secreted by Sertoli cells or germ cells. An interaction network analysis revealed complex, interlaced cell-cell dialog between the secretome and membranome of seminiferous cells, mediated via the TF. We then focused on germ cell-secreted candidate proteins, and we identified several potential interacting partners located on the surface of Sertoli cells. Two interactions, APOH/CDC42 and APP/NGFR, were validated in situ, in a proximity ligation assay (PLA). Our results provide new insight into the crosstalk between germ cells and Sertoli cells occurring during spermatogenesis. Our findings also demonstrate that this "integrative omics" strategy is powerful enough for data mining and highlighting meaningful cell-cell communication events between different types of cells in a complex tissue, via a biological fluid. This integrative strategy could be applied more widely, to gain access to secretomes that have proved difficult to study whilst avoiding the limitations of in vitro culture.

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Integration of “omics” data to establish a network of molecular interactions between germ cells and Sertoli cells mediated by the TF.This integrated network focuses on proteins produced and secreted by germ cells (GCs) or Sertoli cells (SCs) and interacting with membrane proteins of the other type of cell. Nodes symbolizing GC-secreted and Sertoli cell-secreted factors are indicated in light green and purple, respectively, whereas GC-membrane and Sertoli cell-membrane proteins are represented by red and light blue nodes, respectively.
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pone-0104418-g003: Integration of “omics” data to establish a network of molecular interactions between germ cells and Sertoli cells mediated by the TF.This integrated network focuses on proteins produced and secreted by germ cells (GCs) or Sertoli cells (SCs) and interacting with membrane proteins of the other type of cell. Nodes symbolizing GC-secreted and Sertoli cell-secreted factors are indicated in light green and purple, respectively, whereas GC-membrane and Sertoli cell-membrane proteins are represented by red and light blue nodes, respectively.

Mentions: We investigated the extent to which factors secreted by one type of cell (meiotic/post-meiotic germ cells or Sertoli cells) into the TF interacted with cell surface proteins on the other type of cell, by combining cell-specific secretome data and cell-specific membranome data with information about protein-protein interactions. A graphical display created with AMEN [38] revealed a complex interlaced network of interactions between seminiferous cell types, mediated through the TF (Fig. 3). This large network consisted of 22 germ cell-secreted and 23 Sertoli cell-secreted factors interacting with 43 Sertoli cell and 69 germ cell membrane proteins. The physical associations highlighted by this analysis were supported by the findings of 140 published studies on various biological systems and model organisms. This network analysis identified well-known connections between germ cells and somatic cells, such as the intimate association of CLU (clusterin) with SPAM1 (sperm adhesion molecule 1) [39]. CLU is one of the major proteins secreted by Sertoli cells and it has already been associated with the surface of spermatozoa [40]. By contrast, SPAM1 is an important hyaluronidase secreted and acquired by spermatozoa, and playing a key role in fertilization [41], [42].


An integrative omics strategy to assess the germ cell secretome and to decipher sertoli-germ cell crosstalk in the Mammalian testis.

Chalmel F, Com E, Lavigne R, Hernio N, Teixeira-Gomes AP, Dacheux JL, Pineau C - PLoS ONE (2014)

Integration of “omics” data to establish a network of molecular interactions between germ cells and Sertoli cells mediated by the TF.This integrated network focuses on proteins produced and secreted by germ cells (GCs) or Sertoli cells (SCs) and interacting with membrane proteins of the other type of cell. Nodes symbolizing GC-secreted and Sertoli cell-secreted factors are indicated in light green and purple, respectively, whereas GC-membrane and Sertoli cell-membrane proteins are represented by red and light blue nodes, respectively.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4128672&req=5

pone-0104418-g003: Integration of “omics” data to establish a network of molecular interactions between germ cells and Sertoli cells mediated by the TF.This integrated network focuses on proteins produced and secreted by germ cells (GCs) or Sertoli cells (SCs) and interacting with membrane proteins of the other type of cell. Nodes symbolizing GC-secreted and Sertoli cell-secreted factors are indicated in light green and purple, respectively, whereas GC-membrane and Sertoli cell-membrane proteins are represented by red and light blue nodes, respectively.
Mentions: We investigated the extent to which factors secreted by one type of cell (meiotic/post-meiotic germ cells or Sertoli cells) into the TF interacted with cell surface proteins on the other type of cell, by combining cell-specific secretome data and cell-specific membranome data with information about protein-protein interactions. A graphical display created with AMEN [38] revealed a complex interlaced network of interactions between seminiferous cell types, mediated through the TF (Fig. 3). This large network consisted of 22 germ cell-secreted and 23 Sertoli cell-secreted factors interacting with 43 Sertoli cell and 69 germ cell membrane proteins. The physical associations highlighted by this analysis were supported by the findings of 140 published studies on various biological systems and model organisms. This network analysis identified well-known connections between germ cells and somatic cells, such as the intimate association of CLU (clusterin) with SPAM1 (sperm adhesion molecule 1) [39]. CLU is one of the major proteins secreted by Sertoli cells and it has already been associated with the surface of spermatozoa [40]. By contrast, SPAM1 is an important hyaluronidase secreted and acquired by spermatozoa, and playing a key role in fertilization [41], [42].

Bottom Line: Two interactions, APOH/CDC42 and APP/NGFR, were validated in situ, in a proximity ligation assay (PLA).Our findings also demonstrate that this "integrative omics" strategy is powerful enough for data mining and highlighting meaningful cell-cell communication events between different types of cells in a complex tissue, via a biological fluid.This integrative strategy could be applied more widely, to gain access to secretomes that have proved difficult to study whilst avoiding the limitations of in vitro culture.

View Article: PubMed Central - PubMed

Affiliation: IRSET, Inserm U1085, Campus de Beaulieu, Rennes, France.

ABSTRACT
Mammalian spermatogenesis, which takes place in complex testicular structures called seminiferous tubules, is a highly specialized process controlled by the integration of juxtacrine, paracrine and endocrine information. Within the seminiferous tubules, the germ cells and Sertoli cells are surrounded by testicular fluid (TF), which probably contains most of the secreted proteins involved in crosstalk between these cells. It has already been established that germ cells can modulate somatic Sertoli cell function through the secretion of diffusible factors. We studied the germ cell secretome, which was previously considered inaccessible, by analyzing the TF collected by microsurgery in an "integrative omics" strategy combining proteomics, transcriptomics, genomics and interactomics data. This approach identified a set of proteins preferentially secreted by Sertoli cells or germ cells. An interaction network analysis revealed complex, interlaced cell-cell dialog between the secretome and membranome of seminiferous cells, mediated via the TF. We then focused on germ cell-secreted candidate proteins, and we identified several potential interacting partners located on the surface of Sertoli cells. Two interactions, APOH/CDC42 and APP/NGFR, were validated in situ, in a proximity ligation assay (PLA). Our results provide new insight into the crosstalk between germ cells and Sertoli cells occurring during spermatogenesis. Our findings also demonstrate that this "integrative omics" strategy is powerful enough for data mining and highlighting meaningful cell-cell communication events between different types of cells in a complex tissue, via a biological fluid. This integrative strategy could be applied more widely, to gain access to secretomes that have proved difficult to study whilst avoiding the limitations of in vitro culture.

Show MeSH
Related in: MedlinePlus