title: Nodal signaling via an autocrine pathway promotes proliferation of mouse spermatogonial stem/progenitor cells through Smad2/3 and Oct-4 activation.
abstract: Spermatogenesis is the process that involves the division and differentiation of spermatogonial stem cells into spermatozoa. However, the autocrine molecules and signaling pathways controlling their fate remain unknown. This study was designed to identify novel growth factors and signaling pathways that regulate proliferation, differentiation, and survival of spermatogonial stem/progenitor cells. To this end, we have for the first time explored the expression, function, and signaling pathway of Nodal, a member of the transforming growth factor-beta superfamily, in mouse spermatogonial stem/progenitor cells. We demonstrate that both Nodal and its receptors are present in these cells and in a spermatogonial stem/progenitor cell line (C18-4 cells), whereas Nodal is undetected in Sertoli cells or differentiated germ cells, as assayed by reverse transcription-polymerase chain reaction, Western blots, and immunocytochemistry. Nodal promotes proliferation of spermatogonial stem/progenitor cells and C18-4 cells, whereas Nodal receptor inhibitor SB431542 blocks their propagation as shown by proliferation and bromodeoxyuridine incorporation assays. Nodal knockdown by RNA interference results in a marked increase of cell apoptosis and a reduction of cell division as indicated by terminal deoxynucleotidyl transferase dUTP nick-end labeling and proliferation assays. Conversely, overexpression of Nodal leads to an increase of cell proliferation. Nodal activates Smad2/3 phosphorylation, Oct-4 transcription, cyclin D1, and cyclin E expression, whereas SB431542 completely abolishes their increase. Together, Nodal was identified as the first autocrine signaling molecule that promotes proliferation of mouse spermatogonial stem/progenitor cells via Smad2/3 and Oct-4 activation. This study thus provides novel and important insights into molecular mechanisms regulating proliferation and survival of spermatogonial stem/progenitor cells.
authors.affiliation: Department of Biochemistry and Molecular & Cellular Biology, Georgetown University Medical Center, 3900 Reservoir Road NW, Washington, DC 20057, USA.
authors.forename: Zuping
authors.initials: Z
authors.lastname: He
authors.forename: Jiji
authors.initials: J
authors.lastname: Jiang
authors.forename: Maria
authors.initials: M
authors.lastname: Kokkinaki
authors.forename: Martin
authors.initials: M
authors.lastname: Dym
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000235:genetics; D019898:Autocrine Communication / Q000502:physiology; D001549:Benzamides / Q000494:pharmacology; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D049109:Cell Proliferation; D002470:Cell Survival / Q000502:physiology; D016213:Cyclins / Q000378:metabolism; D004149:Dioxoles / Q000494:pharmacology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D055457:Nodal Protein / Q000235:genetics / Q000378:metabolism*; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000037:antagonists & inhibitors / Q000378:metabolism; D051899:Smad2 Protein / Q000235:genetics / Q000378:metabolism*; D013091:Spermatogenesis / Q000502:physiology; D013093:Spermatogonia / Q000166:cytology / Q000378:metabolism*; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism*
chemical_list: C459179:4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamide; D001549:Benzamides; D016213:Cyclins; D004149:Dioxoles; D055457:Nodal Protein; C526008:Nodal protein, mouse; D050814:Octamer Transcription Factor-3; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; D051899:Smad2 Protein; C497783:Smad2 protein, mouse
references.citation: Development. 2005 Mar;132(6):1273-82
references.pmid: 15703277
references.citation: J Cell Biol. 1999 Jan 11;144(1):139-49
references.pmid: 9885250
references.citation: Cell. 2004 Dec 29;119(7):1001-12
references.pmid: 15620358
references.citation: Nature. 2007 Sep 20;449(7160):346-50
references.pmid: 17882221
references.citation: J Biol Chem. 2007 Aug 31;282(35):25842-51
references.pmid: 17597063
references.citation: Curr Biol. 1997 Apr 1;7(4):270-6
references.pmid: 9094310
references.citation: Kidney Int. 2007 May;71(9):867-74
references.pmid: 17332736
references.citation: Reproduction. 2001 Mar;121(3):347-54
references.pmid: 11226060
references.citation: Development. 2007 May;134(10):1853-9
references.pmid: 17428826
references.citation: Nat Genet. 2000 Feb;24(2):157-62
references.pmid: 10655061
references.citation: Dev Biol. 2000 Apr 15;220(2):225-37
references.pmid: 10753512
references.citation: J Androl. 2003 Jan-Feb;24(1):120-30
references.pmid: 12514093
references.citation: Dev Biol. 2007 Apr 1;304(1):34-45
references.pmid: 17222400
references.citation: Biol Reprod. 2004 Jul;71(1):319-30
references.pmid: 15028632
references.citation: Biol Reprod. 2003 Nov;69(5):1565-71
references.pmid: 12855601
references.citation: Proc Natl Acad Sci U S A. 1994 Nov 22;91(24):11287-9
references.pmid: 7972051
references.citation: Dev Biol. 2008 Jan 1;313(1):107-17
references.pmid: 18022151
references.citation: Nature. 2004 Mar 25;428(6981):387-92
references.pmid: 15004567
references.citation: Mol Cell Biol. 2006 Jan;26(2):654-67
references.pmid: 16382155
references.citation: Nature. 2006 Apr 27;440(7088):1199-203
references.pmid: 16565704
references.citation: Science. 2007 Apr 20;316(5823):404-5
references.pmid: 17446391
references.citation: Dev Biol. 2005 Mar 1;279(1):114-24
references.pmid: 15708562
references.citation: Dev Cell. 2007 Feb;12(2):195-206
references.pmid: 17276338
references.citation: J Cell Sci. 2005 Oct 1;118(Pt 19):4495-509
references.pmid: 16179608
references.citation: Genes Dev. 2002 Sep 15;16(18):2339-44
references.pmid: 12231623
references.citation: Science. 2002 Jul 19;297(5580):392-5
references.pmid: 12077424
references.citation: J Cell Sci. 2003 Aug 15;116(Pt 16):3363-72
references.pmid: 12857787
references.citation: Stem Cells. 2008 Nov;26(11):2928-37
references.pmid: 18719224
references.citation: Development. 1994 Jul;120(7):1919-28
references.pmid: 7924997
references.citation: Nat Genet. 2004 Jun;36(6):647-52
references.pmid: 15156142
references.citation: Mol Cell Biol. 1995 May;15(5):2612-24
references.pmid: 7739542
references.citation: Genesis. 2000 Sep;28(1):1-14
references.pmid: 11020711
references.citation: Stem Cells. 2008 Jan;26(1):266-78
references.pmid: 17962702
references.citation: Stem Cells Dev. 2009 Oct;18(8):1115-26
references.pmid: 19281326
references.citation: Nature. 1996 May 9;381(6578):155-8
references.pmid: 8610012
references.citation: Stem Cells. 2005 Feb;23(2):166-85
references.pmid: 15671141
references.citation: Stem Cells. 2005 Feb;23(2):200-10
references.pmid: 15671143
references.citation: Development. 2001 May;128(10):1831-43
references.pmid: 11311163
references.citation: Stem Cells. 2006 Jun;24(6):1505-11
references.pmid: 16456131
references.citation: J Cell Biol. 1977 Jul;74(1):68-85
references.pmid: 874003
references.citation: Stem Cells. 2009 Jan;27(1):138-49
references.pmid: 18927477
references.citation: Biol Reprod. 2007 Oct;77(4):723-33
references.pmid: 17625109
references.citation: Genes Dev. 2003 Jul 1;17(13):1646-62
references.pmid: 12842913
references.citation: Biol Reprod. 1995 Jan;52(1):8-19
references.pmid: 7536046
references.citation: Cancer Res. 2005 Sep 1;65(17):7968-75
references.pmid: 16140969
references.citation: Biol Reprod. 1999 Jul;61(1):225-30
references.pmid: 10377053
references.citation: J Biol Chem. 2004 Oct 22;279(43):45076-84
references.pmid: 15308665
references.citation: Mol Cell Biol. 2004 Nov;24(21):9383-9
references.pmid: 15485907
references.citation: EMBO J. 2000 Mar 15;19(6):1312-26
references.pmid: 10716931
references.citation: J Biol Chem. 2008 May 16;283(20):13992-4001
references.pmid: 18359940
references.citation: Nat Genet. 2000 Apr;24(4):372-6
references.pmid: 10742100
references.citation: Biol Reprod. 2000 Dec;63(6):1893-8
references.pmid: 11090462
references.citation: Mol Pharmacol. 2002 Jul;62(1):65-74
references.pmid: 12065756
references.citation: Biol Reprod. 2006 Feb;74(2):314-21
references.pmid: 16237148
references.citation: Stem Cells. 2008 Apr;26(4):927-38
references.pmid: 18203673
references.citation: Curr Opin Cell Biol. 1998 Dec;10(6):694-701
references.pmid: 9914171
references.citation: Development. 2004 Apr;131(8):1717-28
references.pmid: 15084457
references.citation: Nature. 2008 Nov 20;456(7220):344-9
references.pmid: 18849962
references.citation: J Biol Chem. 2000 Aug 18;275(33):25572-6
references.pmid: 10849422
references.citation: Proc Natl Acad Sci U S A. 2004 Nov 23;101(47):16395-6
references.pmid: 15546980
references.citation: Nature. 1993 Feb 11;361(6412):543-7
references.pmid: 8429908
references.citation: J Cell Sci. 2007 Jan 1;120(Pt 1):55-65
references.pmid: 17182901
references.citation: Science. 2000 Feb 25;287(5457):1489-93
references.pmid: 10688798
references.citation: Proc Natl Acad Sci U S A. 2006 Jun 20;103(25):9524-9
references.pmid: 16740658
references.citation: Dev Biol. 2001 Aug 1;236(1):124-35
references.pmid: 11456449