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Molecular Endocrinology, Vol 10, 439-450, Copyright © 1996 by Endocrine Society

ARTICLES

Steroid and pulsatile gonadotropin-releasing hormone (GnRH) regulation of luteinizing hormone and GnRH receptor in a novel gonadotrope cell line

JL Turgeon, Y Kimura, DW Waring and PL Mellon
Department of Human Physiology, University of California, Davis 95616, USA.

Properties of a pituitary gonadotrope include the capacity to regulate gonadotropin synthesis and secretion in response to a GnRH signal. Progress in identifying the steps involved in these processes has been impeded by the lack of a homogeneous in vitro model of gonadotropes. This paper presents functional characterization of a L beta T2 gonadotrope cell line generated by tumorigenesis in transgenic mice carrying the rat LH beta-subunit regulatory region linked to the SV40 T- antigen oncogene. This cell line expresses LH beta, alpha-subunit, and GnRH-receptor (GnRH-R) mRNAs (though not FSH beta), responds to glucocorticoid treatment with a reversible dampening of proliferation, and responds to pulsatile, concentration-dependent GnRH administration with LH secretion. L beta T2 cells presented with four GnRH pulses (10 nM, 90-min interpulse interval) on each of 4 days respond with incremental increases in LH secretion on successive days. This increase was greatest (15-fold) in the presence of estradiol and dexamethasone. Part of the enhanced responsiveness is apparently due to an increase in GnRH-R; pulsatile GnRH treatment alone as well as steroid treatment alone led to an increase in GnRH-R mRNA levels. When secretion was stimulated on day 4 with 54 mM [K+] pulses, bypassing the GnRH-R, the LH-secretory response indicated that the GnRH pulse history as well as estradiol and dexamethasone have actions on L beta T2-secretory capacity distinct from changes in the GnRH-R. This increase can be explained in part by the marked up-regulation of LH beta, but not alpha- subunit, mRNA observed in GnRH-pulsed cells. In summary, L beta T2 clonal gonadotropes exhibit functional characteristics consistent with those of normal pituitary gonadotropes such as LH secretion via a regulated pathway and changes in GnRH-R and LH beta gene expression in response to signaling by GnRH and steroid hormones and therefore should be a useful tool for dissecting the cellular and molecular events involved in these fundamental gonadotrope properties.


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M. W. Wolfe
The Equine Luteinizing Hormone {beta}-Subunit Promoter Contains Two Functional Steroidogenic Factor-1 Response Elements
Mol. Endocrinol., September 1, 1999; 13(9): 1497 - 1510.
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 EndocrinologyHome page
G. Maya-Núñez and P. M. Conn
Transcriptional Regulation of the Gonadotropin-Releasing Hormone Receptor Gene Is Mediated in Part by a Putative Repressor Element and by the Cyclic Adenosine 3',5'-Monophosphate Response Element
Endocrinology, August 1, 1999; 140(8): 3452 - 3458.
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 EndocrinologyHome page
H. G. Sutton, A. Fusco, and G. A. Cornwall
Cystatin-Related Epididymal Spermatogenic Protein Colocalizes with Luteinizing Hormone-{beta} Protein in Mouse Anterior Pituitary Gonadotropes
Endocrinology, June 1, 1999; 140(6): 2721 - 2732.
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 J. Biol. Chem.Home page
C. Dorn, Q. Ou, J. Svaren, P. A. Crawford, and Y. Sadovsky
Activation of Luteinizing Hormone beta  Gene by Gonadotropin-releasing Hormone Requires the Synergy of Early Growth Response-1 and Steroidogenic Factor-1
J. Biol. Chem., May 14, 1999; 274(20): 13870 - 13876.
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C. T. Albarracin, M. P. Frosch, and W. W. Chin
The Gonadotropin-Releasing Hormone Receptor Gene Promoter Directs Pituitary-Specific Oncogene Expression in Transgenic Mice
Endocrinology, May 1, 1999; 140(5): 2415 - 2421.
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J. J. Tremblay and J. Drouin
Egr-1 Is a Downstream Effector of GnRH and Synergizes by Direct Interaction with Ptx1 and SF-1 To Enhance Luteinizing Hormone beta  Gene Transcription
Mol. Cell. Biol., April 1, 1999; 19(4): 2567 - 2576.
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C.A. Daley, H. Sakurai, B.M. Adams, and T.E. Adams
Effect of Stress-Like Concentrations of Cortisol on Gonadotroph Function in Orchidectomized Sheep
Biol Reprod, January 1, 1999; 60(1): 158 - 163.
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 EndocrinologyHome page
X. Lin and P. M. Conn
Transcriptional Activation of Gonadotropin-Releasing Hormone (GnRH) Receptor Gene by GnRH and Cyclic Adenosine Monophosphate
Endocrinology, September 1, 1998; 139(9): 3896 - 3902.
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T. R. Kumar, K. E. Graham, S. L. Asa, and M. J. Low
Simian Virus 40 T Antigen-Induced Gonadotroph Adenomas: A Model of Human Null Cell Adenomas
Endocrinology, July 1, 1998; 139(7): 3342 - 3351.
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U. B. Kaiser, E. Sabbagh, M. T. Chen, W. W. Chin, and B. D. Saunders
Sp1 Binds to the Rat Luteinizing Hormone beta  (LHbeta ) Gene Promoter and Mediates Gonadotropin-releasing Hormone-stimulated Expression of the LHbeta Subunit Gene
J. Biol. Chem., May 22, 1998; 273(21): 12943 - 12951.
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 Mol. Endocrinol.Home page
A. N. Harris and P. L. Mellon
The Basic Helix-Loop-Helix, Leucine Zipper Transcription Factor, USF (Upstream Stimulatory Factor), Is a Key Regulator of SF-1 (Steroidogenic Factor-1) Gene Expression in Pituitary Gonadotrope and Steroidogenic Cells
Mol. Endocrinol., May 1, 1998; 12(5): 714 - 726.
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U. B. Kaiser, E. Sabbagh, B. D. Saunders, and W. W. Chin
Identification of cis-Acting Deoxyribonucleic Acid Elements That Mediate Gonadotropin-Releasing Hormone Stimulation of the Rat Luteinizing Hormone {beta}-Subunit Gene
Endocrinology, May 1, 1998; 139(5): 2443 - 2451.
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G. A. Cornwall and N. Hsia
ADAM7, A Member of the ADAM (A Disintegrin And Metalloprotease) Gene Family Is Specifically Expressed in the Mouse Anterior Pituitary and Epididymis
Endocrinology, October 1, 1997; 138(10): 4262 - 4272.
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 Endocr. Rev.Home page
U. B. Kaiser, P. M. Conn, and W. W. Chin
Studies of Gonadotropin-Releasing Hormone (GnRH) Action Using GnRH Receptor-Expressing Pituitary Cell Lines
Endocr. Rev., February 1, 1997; 18(1): 46 - 70.
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 DevelopmentHome page
E. Alarid, J. Windle, D. Whyte, and P. Mellon
Immortalization of pituitary cells at discrete stages of development by directed oncogenesis in transgenic mice
Development, January 10, 1996; 122(10): 3319 - 3329.
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T. Yokoi, M. Ohmichi, K. Tasaka, A. Kimura, Y. Kanda, J. Hayakawa, M. Tahara, K. Hisamoto, H. Kurachi, and Y. Murata
Activation of the Luteinizing Hormone beta Promoter by Gonadotropin-releasing Hormone Requires c-Jun NH2-terminal Protein Kinase
J. Biol. Chem., July 7, 2000; 275(28): 21639 - 21647.
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