title: An integrative omics strategy to assess the germ cell secretome and to decipher sertoli-germ cell crosstalk in the Mammalian testis.
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.
authors.affiliation: IRSET, Inserm U1085, Campus de Beaulieu, Rennes, France.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Chalmel
authors.affiliation: Proteomics Core Facility Biogenouest, Inserm U1085 IRSET, Campus de Beaulieu, Rennes, France.
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Com
authors.affiliation: Proteomics Core Facility Biogenouest, Inserm U1085 IRSET, Campus de Beaulieu, Rennes, France.
authors.forename: Régis
authors.initials: R
authors.lastname: Lavigne
authors.affiliation: Proteomics Core Facility Biogenouest, Inserm U1085 IRSET, Campus de Beaulieu, Rennes, France.
authors.forename: Nolwen
authors.initials: N
authors.lastname: Hernio
authors.affiliation: INRA UMR 1282, Infectiologie et Santé Publique, Nouzilly, France; INRA Plate-forme d'Analyse Intégrative des Biomolécules (PAIB2), Nouzilly, France.
authors.forename: Ana-Paula
authors.initials: AP
authors.lastname: Teixeira-Gomes
authors.affiliation: UMR 85 INRA-CNRS, Nouzilly, France.
authors.forename: Jean-Louis
authors.initials: JL
authors.lastname: Dacheux
authors.affiliation: Proteomics Core Facility Biogenouest, Inserm U1085 IRSET, Campus de Beaulieu, Rennes, France.
authors.forename: Charles
authors.initials: C
authors.lastname: Pineau
mesh_terms: D000818:Animals; D002450:Cell Communication; D019295:Computational Biology / Q000379:methods*; D020869:Gene Expression Profiling; D008297:Male; D025941:Protein Interaction Mapping; D040901:Proteomics; D051381:Rats; D012671:Seminiferous Tubules / Q000166:cytology / Q000378:metabolism; D012708:Sertoli Cells / Q000166:cytology*; D013094:Spermatozoa / Q000166:cytology* / Q000378:metabolism*; D013737:Testis / Q000166:cytology*; D014157:Transcription Factors / Q000378:metabolism
chemical_list: D014157:Transcription Factors
references.citation: Nat Commun. 2011 Sep 13;2:472
references.pmid: 21915114
references.citation: J Androl. 2011 Nov-Dec;32(6):651-64
references.pmid: 21764898
references.citation: Mol Cell Proteomics. 2012 Dec;11(12):1682-9
references.pmid: 22949509
references.citation: BMC Bioinformatics. 2008 Feb 06;9:86
references.pmid: 18254954
references.citation: Mol Cell Endocrinol. 1993 May;93(1):53-61
references.pmid: 8319834
references.citation: J Proteome Res. 2008 Sep;7(9):3879-89
references.pmid: 18624398
references.citation: Methods. 2008 Jul;45(3):227-32
references.pmid: 18620061
references.citation: Endocr Rev. 2004 Oct;25(5):747-806
references.pmid: 15466940
references.citation: Biol Reprod. 1994 Mar;50(3):669-79
references.pmid: 8167239
references.citation: Neurodegeneration. 1995 Mar;4(1):51-9
references.pmid: 7600184
references.citation: J Endocrinol. 1988 Sep;118(3):R13-6
references.pmid: 3183569
references.citation: J Reprod Fertil. 1969 Jul;19(2):391-2
references.pmid: 5793479
references.citation: Biol Reprod. 2009 Sep;81(3):562-70
references.pmid: 19357365
references.citation: Int J Androl. 2011 Oct;34(5 Pt 2):e306-18
references.pmid: 21418239
references.citation: J Reprod Fertil. 1981 Mar;61(2):363-71
references.pmid: 7205782
references.citation: J Reprod Fertil. 1979 Nov;57(2):311-8
references.pmid: 574551
references.citation: Biol Reprod. 1995 Feb;52(2):211-6
references.pmid: 7711190
references.citation: Expert Rev Proteomics. 2007 Apr;4(2):239-48
references.pmid: 17425459
references.citation: Nat Protoc. 2013 Nov;8(11):2098-104
references.pmid: 24091557
references.citation: Nucleic Acids Res. 2012 Jan;40(Database issue):D13-25
references.pmid: 22140104
references.citation: Am J Pathol. 1990 Nov;137(5):1027-32
references.pmid: 1700615
references.citation: Mol Cell Endocrinol. 1978 Jan;9(3):335-46
references.pmid: 23974
references.citation: J Reprod Fertil Suppl. 1971 May;13:Suppl 13:15-27
references.pmid: 5292797
references.citation: Biol Reprod. 2004 Jul;71(1):319-30
references.pmid: 15028632
references.citation: PLoS One. 2013 Dec 16;8(12):e82321
references.pmid: 24358169
references.citation: Nucleic Acids Res. 2011 Jan;39(Database issue):D289-94
references.pmid: 21113020
references.citation: Nucleic Acids Res. 2010 Jan;38(Database issue):D736-42
references.pmid: 19906717
references.citation: Nucleic Acids Res. 2005 Jan 1;33(Database issue):D169-73
references.pmid: 15608170
references.citation: Int Rev Cell Mol Biol. 2009;278:309-53
references.pmid: 19815182
references.citation: Int J Mol Med. 2006 Mar;17(3):523-8
references.pmid: 16465402
references.citation: Syst Biol Reprod Med. 2012 Aug;58(4):179-90
references.pmid: 22788530
references.citation: Nucleic Acids Res. 2009 Jan;37(Database issue):D767-72
references.pmid: 18988627
references.citation: J Proteome Res. 2014 Mar 7;13(3):1259-69
references.pmid: 24422521
references.citation: J Androl. 1993 Mar-Apr;14(2):87-98
references.pmid: 8514623
references.citation: Recent Prog Horm Res. 2002;57:103-28
references.pmid: 12017539
references.citation: J Reprod Fertil. 1983 Sep;69(1):125-35
references.pmid: 6136607
references.citation: J Reprod Fertil Suppl. 1979;(26):97-116
references.pmid: 42791
references.citation: Nature. 2011 Mar 24;471(7339):504-7
references.pmid: 21430778
references.citation: Endocrinology. 1989 Jun;124(6):2720-8
references.pmid: 2656244
references.citation: Cell Rep. 2013 Jun 27;3(6):2179-90
references.pmid: 23791531
references.citation: Ann N Y Acad Sci. 2007 Jan;1096:196-206
references.pmid: 17405931
references.citation: Hum Reprod. 2013 Jan;28(1):199-209
references.pmid: 23024119
references.citation: Proteins. 2006 Mar 15;62(3):776-92
references.pmid: 16342272
references.citation: Int Rev Cytol. 1993;147:25-96
references.pmid: 8225836
references.citation: Reproduction. 2005 Jan;129(1):1-7
references.pmid: 15615893
references.citation: Physiol Rev. 1988 Apr;68(2):555-607
references.pmid: 3282246
references.citation: J Reprod Fertil Suppl. 1979;(26):165-73
references.pmid: 293407
references.citation: Philos Trans R Soc Lond B Biol Sci. 2010 May 27;365(1546):1481-500
references.pmid: 20403865
references.citation: Nucleic Acids Res. 2012 Jan;40(Database issue):D857-61
references.pmid: 22096227
references.citation: Proc Natl Acad Sci U S A. 2007 May 15;104(20):8346-51
references.pmid: 17483452
references.citation: Biochim Biophys Acta. 2013 Nov;1834(11):2454-61
references.pmid: 23603790
references.citation: Physiol Rev. 1972 Jan;52(1):198-236
references.pmid: 4621362
references.citation: J Cell Biol. 2003 Mar 3;160(5):629-33
references.pmid: 12615908
references.citation: Mol Cell Proteomics. 2012 Feb;11(2):M111.012682
references.pmid: 21997732
references.citation: Electrophoresis. 2005 Jul;26(14):2779-85
references.pmid: 15966010
references.citation: Nucleic Acids Res. 2012 Jan;40(Database issue):D841-6
references.pmid: 22121220
references.citation: Nature. 2009 Feb 19;457(7232):981-9
references.pmid: 19225519
references.citation: Proc Natl Acad Sci U S A. 2010 Jun 22;107(25):11399-404
references.pmid: 20534521
references.citation: Expert Rev Proteomics. 2012 Jun;9(3):337-45
references.pmid: 22809211
references.citation: J Cell Physiol. 2007 Jan;210(1):51-62
references.pmid: 17013810
references.citation: J Reprod Fertil. 1967 Oct;14(2):347-8
references.pmid: 6053782
references.citation: J Biol Chem. 1995 Oct 27;270(43):25845-9
references.pmid: 7592769
references.citation: Nat Cell Biol. 2002 Oct;4 Suppl:s41-9
references.pmid: 12479614
references.citation: Nucleic Acids Res. 2011 Jan;39(Database issue):D698-704
references.pmid: 21071413
references.citation: Mol Cell Proteomics. 2003 Apr;2(4):248-61
references.pmid: 12754304
references.citation: Ann N Y Acad Sci. 2007 Dec;1120:1-15
references.pmid: 17911412
references.citation: Biotechniques. 2011 Aug;51(2):111-8
references.pmid: 21806555
references.citation: Reproduction. 2001 Mar;121(3):347-54
references.pmid: 11226060
references.citation: Curr Opin Biotechnol. 2010 Oct;21(5):621-6
references.pmid: 20637599
references.citation: Mol Cell Endocrinol. 2006 May 16;250(1-2):114-21
references.pmid: 16420970
references.citation: Prog Clin Biol Res. 1982;87:193-216
references.pmid: 7051031
references.citation: Biosci Rep. 2002 Apr;22(2):197-224
references.pmid: 12428901
references.citation: Mol Reprod Dev. 2006 Jul;73(7):866-77
references.pmid: 16557521
references.citation: Nat Rev Drug Discov. 2007 Jul;6(7):569-82
references.pmid: 17599086
references.citation: Reproduction. 2013 Apr 15;145(4):R89-95
references.pmid: 23404847
references.citation: Cytogenet Genome Res. 2003;103(3-4):267-76
references.pmid: 15051947
references.citation: Curr Opin Biotechnol. 2003 Feb;14(1):82-6
references.pmid: 12566006
references.citation: Adv Exp Med Biol. 2008;636:16-41
references.pmid: 19856160