title: TGF-β1 regulation of estrogen production in mature rat Leydig cells.
abstract: BACKGROUND
  Besides androgens, estrogens produced in Leydig cells are also crucial for mammalian germ cell differentiation. Transforming growth factor-β1 (TGF-β1) is now known to have multiple effects on regulation of Leydig cell function. The objective of the present study is to determine whether TGF-β1 regulates estradiol (E2) synthesis in adult rat Leydig cells and then to assess the impact of TGF-β1 on Cx43-based gap junctional intercellular communication (GJIC) between Leydig cells.
  
  
  RESULTS
  Primary cultured Leydig cells were incubated in the presence of recombinant TGF-β1 and the production of E2 as well as testosterone (T) were measured by RIA. The activity of P450arom was addressed by the tritiated water release assay and the expression of Cyp19 gene was evaluated by Western blotting and real time RT-PCR. The expression of Cx43 and GJIC were investigated with immunofluorescence and fluorescence recovery after photo-bleaching (FRAP), respectively. Results from this study show that TGF-β1 down-regulates the level of E2 secretion and the activity of P450arom in a dose-dependent manner in adult Leydig cells. In addition, the expression of Cx43 and GJIC was closely related to the regulation of E2 and TGF-β1, and E2 treatment in turn restored the inhibition of TGF-β1 on GJIC.
  
  
  CONCLUSIONS
  Our results indicate, for the first time in adult rat Leydig cells, that TGF-β1 suppresses P450arom activity, as well as the expression of the Cyp19 gene, and that depression of E2 secretion leads to down-regulation of Cx43-based GJIC between Leydig cells.
authors.affiliation: Department of Traditional Chinese Medicine, Xijing Hospital, the Fourth Military Medical University, Xi'an, People's Republic of China.
authors.forename: Man-Li
authors.initials: ML
authors.lastname: Liu
authors.forename: Huan
authors.initials: H
authors.lastname: Wang
authors.forename: Zong-Ren
authors.initials: ZR
authors.lastname: Wang
authors.forename: Yu-Fen
authors.initials: YF
authors.lastname: Zhang
authors.forename: Yan-Qiu
authors.initials: YQ
authors.lastname: Chen
authors.forename: Fang-Hong
authors.initials: FH
authors.lastname: Zhu
authors.forename: Yuan-Qiang
authors.initials: YQ
authors.lastname: Zhang
authors.forename: Jing
authors.initials: J
authors.lastname: Ma
authors.forename: Zhen
authors.initials: Z
authors.lastname: Li
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D004967:Estrogens / Q000378:metabolism* / Q000494:pharmacology; D017629:Gap Junctions / Q000187:drug effects / Q000378:metabolism; D007985:Leydig Cells / Q000187:drug effects* / Q000378:metabolism*; D008297:Male; D016133:Polymerase Chain Reaction; D011863:Radioimmunoassay; D051381:Rats; D017207:Rats, Sprague-Dawley; D013739:Testosterone / Q000378:metabolism; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology*
chemical_list: D004967:Estrogens; D053773:Transforming Growth Factor beta1; D013739:Testosterone
references.citation: J Steroid Biochem. 1988;30(1-6):443-7
references.pmid: 3164434
references.citation: Mol Hum Reprod. 2001 Jul;7(7):671-9
references.pmid: 11420391
references.citation: Int J Androl. 2006 Feb;29(1):90-5; discussion 105-8
references.pmid: 16466528
references.citation: Endocrinology. 1995 Dec;136(12):5322-8
references.pmid: 7588277
references.citation: Reprod Biol Endocrinol. 2003 Jul 09;1:52
references.pmid: 12904263
references.citation: Biol Reprod. 2003 Aug;69(2):592-601
references.pmid: 12700195
references.citation: Trends Endocrinol Metab. 2005 Sep;16(7):300-6
references.pmid: 16054834
references.citation: Cell. 1992 Feb 21;68(4):731-42
references.pmid: 1739978
references.citation: Eur J Endocrinol. 2012 May;166(5):941-9
references.pmid: 22301800
references.citation: Biol Reprod. 2001 May;64(5):1439-43
references.pmid: 11319149
references.citation: Biol Reprod. 2004 Jul;71(1):273-81
references.pmid: 15028626
references.citation: Reproduction. 2008 Oct;136(4):447-57
references.pmid: 18635743
references.citation: J Steroid Biochem Mol Biol. 1998 Aug;66(3):105-12
references.pmid: 9719444
references.citation: Mol Cell Endocrinol. 1998 Apr 30;139(1-2):117-29
references.pmid: 9705080
references.citation: J Reprod Dev. 2010 Aug;56(4):400-4
references.pmid: 20431251
references.citation: ILAR J. 2004;45(4):455-61
references.pmid: 15454684
references.citation: J Cardiovasc Pharmacol. 2002 Dec;40(6):938-48
references.pmid: 12451328
references.citation: Physiol Rev. 2003 Oct;83(4):1359-400
references.pmid: 14506308
references.citation: Reprod Biol Endocrinol. 2003 Apr 11;1:35
references.pmid: 12747806
references.citation: J Steroid Biochem Mol Biol. 2003 Sep;86(3-5):219-24
references.pmid: 14623514
references.citation: Mol Cell Endocrinol. 2000 Feb 25;160(1-2):123-33
references.pmid: 10715546
references.citation: Reprod Biol Endocrinol. 2009 Dec 09;7:146
references.pmid: 20003198
references.citation: Endocrinology. 2004 May;145(5):2186-96
references.pmid: 14736734
references.citation: Mol Cell Endocrinol. 2002 Sep 30;195(1-2):65-77
references.pmid: 12354673
references.citation: Mol Endocrinol. 2003 May;17(5):818-30
references.pmid: 12554767
references.citation: Endocr Res. 2005;31(2):71-9
references.pmid: 16353668
references.citation: Gut. 1968 Feb;9(1):42-5
references.pmid: 5640925
references.citation: J Steroid Biochem Mol Biol. 2001 Dec;79(1-5):203-8
references.pmid: 11850226
references.citation: Mol Hum Reprod. 2002 Apr;8(4):318-25
references.pmid: 11912279
references.citation: Mol Cell Endocrinol. 1999 Feb 25;148(1-2):21-8
references.pmid: 10221767
references.citation: Mol Cell Biol. 2010 Jan;30(1):206-19
references.pmid: 19841066
references.citation: J Cell Physiol. 2004 Jul;200(1):146-54
references.pmid: 15137067
references.citation: Proc Soc Exp Biol Med. 1991 Mar;196(3):351-8
references.pmid: 1671798
references.citation: J Mol Cell Cardiol. 2004 Nov;37(5):1013-22
references.pmid: 15522278
references.citation: Endocrinology. 2001 Jan;142(1):359-69
references.pmid: 11145599
references.citation: Mol Cell Endocrinol. 2001 Jun 10;178(1-2):141-6
references.pmid: 11403903
references.citation: Gene. 2002 Jul 10;294(1-2):279-90
references.pmid: 12234690
references.citation: N Engl J Med. 1994 Oct 20;331(16):1056-61
references.pmid: 8090165
references.citation: Proc Biol Sci. 1994 Feb 22;255(1343):125-32
references.pmid: 8165225
references.citation: Biol Cell. 2012 Jul;104(7):378-96
references.pmid: 22420773
references.citation: J Steroid Biochem Mol Biol. 2008 Apr;109(3-5):212-8
references.pmid: 18448329
references.citation: Cell Tissue Res. 2008 Nov;334(2):283-94
references.pmid: 18795335
references.citation: Mol Cell Endocrinol. 2010 Jul 29;323(2):307-13
references.pmid: 20214950
references.citation: J Steroid Biochem Mol Biol. 2003 Sep;86(3-5):335-43
references.pmid: 14623530
references.citation: Mol Cell Biochem. 2008 Jan;307(1-2):65-71
references.pmid: 17786540
references.citation: Steroids. 1999 Sep;64(9):610-7
references.pmid: 10503717
references.citation: J Steroid Biochem Mol Biol. 1999 Apr-Jun;69(1-6):379-84
references.pmid: 10419015
references.citation: J Endocrinol. 1992 May;133(2):311-20
references.pmid: 1613432
references.citation: Int J Biochem Cell Biol. 2009 Nov;41(11):2323-33
references.pmid: 19523531
references.citation: Hum Reprod. 1999 Jun;14(6):1534-9
references.pmid: 10357971
references.citation: Endocr Rev. 2001 Jun;22(3):289-318
references.pmid: 11399746
references.citation: J Endocrinol. 2000 Aug;166(2):447-53
references.pmid: 10927634
references.citation: Int J Biochem Cell Biol. 2009 Jun;41(6):1381-90
references.pmid: 19136074