title: Identification of androgen receptor phosphorylation in the primate ovary in vivo.
abstract: The androgen receptor (AR) is a member of the nuclear receptor superfamily, and is important for both male and female reproductive health. The receptor is a target for a number of post-translational modifications including phosphorylation, which has been intensively studied in vitro. However, little is known about the phosphorylation status of the receptor in target tissues in vivo. The common marmoset is a useful model for studying human reproductive functions, and comparison of the AR primary sequence from this primate shows high conservation of serines known to be phosphorylated in the human receptor and corresponding flanking amino acids. We have used a panel of phosphospecific antibodies to study AR phosphorylation in the marmoset ovary throughout the follicular phase and after treatment with GNRH antagonist or testosterone propionate. In normal follicular phase ovaries, total AR (both phosphorylated and non-phosphorylated forms) immunopositive staining was observed in several cell types including granulosa cells of developing follicles, theca cells and endothelial cells lining blood vessels. Receptor phosphorylation at serines 81, 308, and 650 was detected primarily in the granulosa cells of developing follicles, surface epithelium, and vessel endothelial cells. Testosterone treatment lead to a modest increase in AR staining in all stages of follicle studied, while GNRH antagonist had no effect. Neither treatment significantly altered the pattern of phosphorylation compared to the control group. These results demonstrate that phosphorylation of the AR occurs, at a subset of serine residues, in a reproductive target tissue in vivo, which appears refractory to hormonal manipulations.
authors.affiliation: MRC Human Reproductive Sciences Unit, Queen's Medical Research Institute, 47 Little France Crescent, Edinburgh EH16 4TJ, Scotland, UK. iain.mcewan@abdn.ac.uk
authors.forename: Iain J
authors.initials: IJ
authors.lastname: McEwan
authors.forename: Dagmara
authors.initials: D
authors.lastname: McGuinness
authors.forename: Colin W
authors.initials: CW
authors.lastname: Hay
authors.forename: Robert P
authors.initials: RP
authors.lastname: Millar
authors.forename: Philippa T K
authors.initials: PT
authors.lastname: Saunders
authors.forename: Hamish M
authors.initials: HM
authors.lastname: Fraser
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002144:Callithrix / Q000502:physiology*; D042783:Endothelial Cells / Q000502:physiology; D005260:Female; D005498:Follicular Phase / Q000502:physiology; D006107:Granulosa Cells / Q000502:physiology; D007150:Immunohistochemistry; D008297:Male; D008969:Molecular Sequence Data; D006080:Ovarian Follicle / Q000166:cytology / Q000502:physiology; D010053:Ovary / Q000166:cytology / Q000254:growth & development / Q000378:metabolism*; D010766:Phosphorylation; D010768:Phosphoserine / Q000378:metabolism; D011944:Receptors, Androgen / Q000378:metabolism*; D011966:Receptors, LHRH / Q000037:antagonists & inhibitors; D013739:Testosterone / Q000097:blood / Q000494:pharmacology / Q000502:physiology
chemical_list: D011944:Receptors, Androgen; D011966:Receptors, LHRH; D010768:Phosphoserine; D013739:Testosterone
references.citation: Bioinformatics. 2007 Nov 1;23(21):2947-8
references.pmid: 17846036
references.citation: Mol Cell Biol. 2002 May;22(10):3373-88
references.pmid: 11971970
references.citation: J Clin Endocrinol Metab. 1999 Aug;84(8):2951-6
references.pmid: 10443703
references.citation: Endocrinology. 1996 Oct;137(10):4392-7
references.pmid: 8828500
references.citation: Mol Hum Reprod. 2007 Oct;13(10):729-36
references.pmid: 17804434
references.citation: Reprod Fertil Dev. 2009;21(3):377-92
references.pmid: 19261215
references.citation: EMBO J. 2002 Aug 1;21(15):4037-48
references.pmid: 12145204
references.citation: Mol Endocrinol. 2004 Apr;18(4):834-50
references.pmid: 14684849
references.citation: Mol Cell Biol. 2005 Feb;25(4):1298-308
references.pmid: 15684382
references.citation: Proc Natl Acad Sci U S A. 2006 Oct 24;103(43):15969-74
references.pmid: 17043241
references.citation: J Biol Chem. 2003 Nov 14;278(46):46087-93
references.pmid: 12958311
references.citation: Proc Natl Acad Sci U S A. 2006 Jan 3;103(1):224-9
references.pmid: 16373508
references.citation: J Biol Chem. 2002 Aug 9;277(32):29304-14
references.pmid: 12015328
references.citation: Proc Natl Acad Sci U S A. 2004 Aug 3;101(31):11209-14
references.pmid: 15277682
references.citation: Endocrinology. 1999 Oct;140(10):4753-60
references.pmid: 10499535
references.citation: Clin Endocrinol (Oxf). 2008 Apr;68(4):580-8
references.pmid: 17970778
references.citation: J Biol Chem. 2008 Jul 25;283(30):20989-1001
references.pmid: 18511414
references.citation: J Clin Oncol. 2002 Jul 1;20(13):3001-15
references.pmid: 12089231
references.citation: Endocr Relat Cancer. 2004 Jun;11(2):281-93
references.pmid: 15163303
references.citation: Biochemistry. 1998 Mar 17;37(11):3850-7
references.pmid: 9521705
references.citation: Biol Reprod. 2005 Oct;73(4):825-32
references.pmid: 15972887
references.citation: Mol Endocrinol. 1995 May;9(5):605-15
references.pmid: 7565807
references.citation: J Biol Chem. 2005 Dec 9;280(49):40916-24
references.pmid: 16210317
references.citation: J Endocrinol. 1995 Jun;145(3):535-43
references.pmid: 7636438
references.citation: Biol Reprod. 2000 Oct;63(4):1098-105
references.pmid: 10993832
references.citation: J Clin Endocrinol Metab. 1998 Jul;83(7):2479-85
references.pmid: 9661631
references.citation: Trends Endocrinol Metab. 2007 Jul;18(5):183-9
references.pmid: 17442585
references.citation: J Endocrinol. 2004 Oct;183(1):1-17
references.pmid: 15525569
references.citation: Biochem J. 2004 Oct 15;383(Pt 2):267-76
references.pmid: 15239671
references.citation: J Biol Chem. 2009 Sep 18;284(38):25576-84
references.pmid: 19622840
references.citation: Mol Endocrinol. 2006 Mar;20(3):503-15
references.pmid: 16282370
references.citation: Proc Natl Acad Sci U S A. 2000 Dec 19;97(26):14145-50
references.pmid: 11121022
references.citation: J Biol Chem. 2002 Jul 19;277(29):25904-13
references.pmid: 11994312
references.citation: Endocrinology. 2007 May;148(5):2273-81
references.pmid: 17317775
references.citation: Biochemistry. 1994 Nov 29;33(47):14064-72
references.pmid: 7947816
references.citation: Endocr Rev. 1997 Feb;18(1):71-106
references.pmid: 9034787
references.citation: J Endocrinol. 2000 May;165(2):359-70
references.pmid: 10810300
references.citation: J Clin Endocrinol Metab. 2007 Mar;92(3):1034-40
references.pmid: 17179193
references.citation: Mol Cell Biol. 2007 Oct;27(20):7125-42
references.pmid: 17698582
references.citation: Mol Cell Endocrinol. 2001 Jun 20;179(1-2):39-46
references.pmid: 11420129
references.citation: Proc Natl Acad Sci U S A. 2001 Jun 19;98(13):7200-5
references.pmid: 11404460
references.citation: Mol Endocrinol. 2004 Jun;18(6):1438-49
references.pmid: 15031320
references.citation: J Clin Endocrinol Metab. 1997 Mar;82(3):955-62
references.pmid: 9062513
references.citation: Hum Pathol. 1994 Nov;25(11):1198-204
references.pmid: 7959665
references.citation: Biol Reprod. 2001 Jan;64(1):127-35
references.pmid: 11133667
references.citation: Br J Cancer. 2008 Mar 25;98(6):1094-101
references.pmid: 18349820
references.citation: Br J Cancer. 2002 Mar 18;86(6):879-85
references.pmid: 11953818
references.citation: Clin Endocrinol (Oxf). 2006 Nov;65(5):606-10
references.pmid: 17054461
references.citation: Proc Natl Acad Sci U S A. 2007 May 15;104(20):8438-43
references.pmid: 17494760
references.citation: Endocrinology. 2007 Aug;148(8):3674-84
references.pmid: 17463055
references.citation: Biol Reprod. 2009 Apr;80(4):685-95
references.pmid: 19074005
references.citation: Reproduction. 2006 Jan;131(1):81-92
references.pmid: 16388012
references.citation: J Biol Chem. 1991 Feb 25;266(6):3734-8
references.pmid: 1995627
references.citation: Hum Reprod. 1997 Jan;12(1):107-11
references.pmid: 9043913
references.citation: Hum Reprod. 1992 Nov;7(10):1461-6
references.pmid: 1291578
references.citation: Mol Cell Biol. 2007 May;27(9):3390-404
references.pmid: 17325038
references.citation: Mol Endocrinol. 2007 Sep;21(9):2071-84
references.pmid: 17579212
references.citation: Hum Reprod. 1999 Sep;14(9):2328-32
references.pmid: 10469704
references.citation: Proc Natl Acad Sci U S A. 2002 Oct 15;99(21):13498-503
references.pmid: 12370412
references.citation: Cell Cycle. 2006 Jun;5(11):1178-83
references.pmid: 16760656
references.citation: Biochem Biophys Res Commun. 2001 Jun 15;284(3):836-44
references.pmid: 11396978
references.citation: J Cereb Blood Flow Metab. 2007 Nov;27(11):1841-52
references.pmid: 17406656