title: GnRH pulse frequency-dependent stimulation of FSHβ transcription is mediated via activation of PKA and CREB.
abstract: Expression of pituitary FSH and LH, under the control of pulsatile GnRH, is essential for fertility. cAMP response element-binding protein (CREB) has been implicated in the regulation of FSHβ gene expression, but the molecular mechanisms by which pulsatile GnRH regulates CREB activation remain poorly understood. We hypothesized that CREB is activated by a distinct signaling pathway in response to pulsatile GnRH in a frequency-dependent manner to dictate the FSHβ transcriptional response. GnRH stimulation of CREB phosphorylation (pCREB) in the gonadotrope-derived LβT2 cell line was attenuated by a protein kinase A (PKA) inhibitor, H89. A dominant negative PKA (DNPKA) reduced GnRH-stimulated pCREB and markedly decreased GnRH stimulation of FSHβ mRNA and FSHβLUC activity, but had little effect on LHβLUC activity, indicating relative specificity of this pathway. In perifusion studies, FSHβ mRNA levels and FSHβLUC activities were increased by pulsatile GnRH, with significantly greater increases at low compared with high pulse frequencies. DNPKA markedly reduced these GnRH-stimulated FSHβ responses at both low and high pulse frequencies. Correlating with FSHβ activation, both PKA activity and levels of pCREB were increased to a greater extent by low compared with high GnRH pulse frequencies, and the induction of pCREB was also attenuated by overexpression of DNPKA at both low and high pulse frequencies. Taken together, these data indicate that a PKA-mediated signaling pathway mediates GnRH activation of CREB at low-pulse frequencies, playing a significant role in the decoding of the hypothalamic GnRH signal to result in frequency-dependent FSHβ activation.
authors: Division of Endocrinology, Diabetes and Hypertension, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA.
authors: Iain R
authors: IR
authors: Thompson
authors: Nick A
authors: NA
authors: Ciccone
authors: Shuyun
authors: S
authors: Xu
authors: Sofiya
authors: S
authors: Zaytseva
authors: Rona S
authors: RS
authors: Carroll
authors: Ursula B
authors: UB
authors: Kaiser
mesh_terms: D000818:Animals; D002460:Cell Line; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects; D037201:Follicle Stimulating Hormone, beta Subunit / Q000235:genetics*; D005786:Gene Expression Regulation / Q000187:drug effects; D005799:Genes, Dominant; D007987:Gonadotropin-Releasing Hormone / Q000494:pharmacology*; D051379:Mice; D010766:Phosphorylation / Q000187:drug effects; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects*
chemical_list: D017362:Cyclic AMP Response Element-Binding Protein; D037201:Follicle Stimulating Hormone, beta Subunit; D047428:Protein Kinase Inhibitors; D012333:RNA, Messenger; D007987:Gonadotropin-Releasing Hormone; D017868:Cyclic AMP-Dependent Protein Kinases
references: Endocrinology. 2003 Jun;144(6):2409-16
references: 12746302
references: J Biol Chem. 2009 Dec 18;284(51):35746-57
references: 19858197
references: Endocr Rev. 1998 Oct;19(5):521-39
references: 9793755
references: Mol Endocrinol. 2002 Mar;16(3):419-34
references: 11875099
references: Mol Endocrinol. 1998 Jan;12(1):107-22
references: 9440815
references: J Biol Chem. 2004 Jan 2;279(1):152-62
references: 14570911
references: J Biol Chem. 2000 May 12;275(19):14182-9
references: 10799494
references: Proc Natl Acad Sci U S A. 1995 Dec 19;92(26):12280-4
references: 8618885
references: J Biol Chem. 1998 Jun 12;273(24):14712-20
references: 9614069
references: Endocrinology. 2008 Nov;149(11):5577-91
references: 18653705
references: Front Neuroendocrinol. 1998 Jan;19(1):1-19
references: 9465287
references: Development. 1996 Oct;122(10):3319-29
references: 8898243
references: Biol Reprod. 2010 Jun;82(6):1170-9
references: 20181617
references: Mol Endocrinol. 1999 Aug;13(8):1338-52
references: 10446907
references: Nature. 1977 Sep 22;269(5626):338-40
references: 198666
references: Comp Biochem Physiol B Biochem Mol Biol. 2001 Jun;129(2-3):525-32
references: 11399488
references: Endocrinology. 2002 Sep;143(9):3243-9
references: 12193535
references: Mol Endocrinol. 2002 Aug;16(8):1840-52
references: 12145338
references: Proc Natl Acad Sci U S A. 1972 Sep;69(9):2677-81
references: 4341705
references: Science. 1996 Aug 30;273(5279):1219-21
references: 8703054
references: Biochem Soc Trans. 2012 Feb;40(1):273-8
references: 22260704
references: J Neuroendocrinol. 1995 Oct;7(10):739-57
references: 8563717
references: Endocrinology. 1997 Mar;138(3):1224-31
references: 9048630
references: Endocrinology. 1990 Nov;127(5):2364-71
references: 2121462
references: J Biol Chem. 2000 Jul 14;275(28):21639-47
references: 10787426
references: Mol Endocrinol. 1999 May;13(5):752-63
references: 10319325
references: Mol Cell Biol. 1999 Apr;19(4):2567-76
references: 10082522
references: Gen Comp Endocrinol. 2008 Aug;158(1):36-46
references: 18558406
references: J Biol Chem. 2010 Aug 6;285(32):24360-71
references: 20507982
references: Mol Endocrinol. 2000 Aug;14(8):1235-45
references: 10935547
references: Endocr Rev. 1990 Feb;11(1):177-99
references: 2108012
references: Adv Exp Med Biol. 1987;219:641-5
references: 3124522
references: Mol Endocrinol. 1998 Mar;12(3):451-7
references: 9514161
references: Mol Endocrinol. 2008 Aug;22(8):1908-23
references: 18550775
references: Endocrinology. 2003 May;144(5):1802-11
references: 12697686
references: Mol Endocrinol. 2006 Nov;20(11):2909-30
references: 16772531
references: J Clin Endocrinol Metab. 1991 Mar;72(3):711-7
references: 1997524
references: Mol Endocrinol. 2007 Dec;21(12):3071-86
references: 17823303
references: Mol Endocrinol. 1999 Jan;13(1):106-16
references: 9892016
references: Mol Endocrinol. 2009 Jul;23(7):1022-32
references: 19342443
references: Biol Reprod. 2008 Nov;79(5):947-53
references: 18716286
references: Endocrinology. 1997 Jun;138(6):2259-66
references: 9165009
references: J Biol Chem. 1989 Aug 5;264(22):13321-8
references: 2546952
references: J Biol Chem. 1999 May 14;274(20):13870-6
references: 10318795
references: Mol Endocrinol. 2009 Jan;23(1):100-12
references: 18974261
references: Nature. 1994 Jul 21;370(6486):223-6
references: 7913207
references: Mol Cell Biol. 2010 Feb;30(4):1028-40
references: 20008557
references: Biol Reprod. 2009 Dec;81(6):1206-15
references: 19710510
references: J Mol Endocrinol. 2004 Dec;33(3):559-84
references: 15591020
references: Biol Reprod. 2001 Dec;65(6):1789-93
references: 11717142
references: Mol Endocrinol. 2011 Aug;25(8):1387-403
references: 21659477
references: Endocrinology. 1991 Jan;128(1):509-17
references: 1702704
references: Endocrinology. 2005 Dec;146(12):5503-13
references: 16141398
references: J R Soc Interface. 2012 Jan 7;9(66):170-82
references: 21676968
references: Nat Clin Pract Endocrinol Metab. 2008 Apr;4(4):215-22
references: 18250636
references: Biol Reprod. 1999 Sep;61(3):715-23
references: 10456849
references: Semin Reprod Med. 2007 Sep;25(5):352-9
references: 17710731
references: Endocrinology. 2003 Jul;144(7):2768-74
references: 12810529
references: J Biol Chem. 2010 Jun 25;285(26):20262-72
references: 20406815
references: Endocrinology. 1989 Aug;125(2):917-24
references: 2502379
references: Mol Cell Endocrinol. 2007 Jun 15;271(1-2):45-54
references: 17482756
references: Biol Reprod. 1989 Sep;41(3):454-63
references: 2511933
references: N Engl J Med. 2005 Mar 24;352(12):1223-36
references: 15788499
references: Mol Endocrinol. 2002 Feb;16(2):221-33
references: 11818496
references: Endocrinology. 1993 Aug;133(2):931-4
references: 8393779
references: J Biol Chem. 1999 Oct 15;274(42):29796-804
references: 10514457
references: Endocrinology. 2008 Jan;149(1):139-45
references: 17932215
references: Endocr Rev. 1997 Dec;18(6):774-800
references: 9408743
references: Endocrinology. 1981 Aug;109(2):376-85
references: 6788538
references: Recent Prog Horm Res. 1995;50:161-205
references: 7740156
references: Mol Endocrinol. 2007 May;21(5):1175-91
references: 17299135
references: J Biol Chem. 2002 Aug 30;277(35):32099-108
references: 12050161
references: Mol Endocrinol. 1990 Apr;4(4):597-603
references: 1704103
references: Endocrinology. 1983 Aug;113(2):535-44
references: 6409585
references: Endocrinology. 2007 Mar;148(3):1099-107
references: 17185372
references: Endocrinology. 2003 Feb;144(2):612-22
references: 12538624