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Studies of Gonadotropin-Releasing Hormone (GnRH) Action Using GnRH Receptor-Expressing Pituitary Cell Lines¹

Division of Genetics (U.B.K., W.W.C.), Department of Medicine, Brigham and Women’s Hospital, Harvard Medical School, Boston, Massachusetts 02115; Oregon Regional Primate Research Center (P.M.C.), Beaverton, Oregon 97006; and Oregon Health Sciences University, Portland, Oregon 97201-3098

I. Introduction

II. GnRHR Structure Analysis

III. Studies of GnRH Action in T3-1 Cells

A. Derivation of T3-1 cells

B. Characterization of T3-1 cells

C. GnRH binding

D. GnRHR regulation

1. Homologous regulation by GnRH

2. Regulation by gonadal steroid hormones

3. Regulation by gonadal peptides

4. Regulation by second messenger activators

E. Intracellular second messengers

1. G protein coupling

2. Inositol phosphates

3. Intracellular calcium

4. Protein kinase C

5. cAMP

6. Mitogen-activated protein kinases

F. -Subunit gene expression

1. Cell-specific expression

2. GnRH-stimulated expression

G. Desensitization

H. Summary of GnRH action in T3-1 cells

IV. Studies of GnRH Action in GH₃ Cells Transfected with the GnRH Receptor (GGH₃ Cells)

A. Derivation of GH₃ cells

B. Characterization of GH₃ cells

C. Derivation of GH₃ cells transfected with the GnRHR (GGH₃ cells)

D. GnRH binding

E. GnRHR regulation

F. Intracellular second messengers

1. G protein coupling

2. Inositol phosphates

3. cAMP

G. Regulation of secretion

1. PRL

2. LH and FSH (in GH₃ cells transfected with the -, LHß-, and FSHß-subunit genes)

3. Secretogranin-II

H. Regulation of PRL mRNA

I. Regulation of expression of transiently expressed reporter genes

1. PRL vs. -subunit gene

2. -, LHß-, and FSHß-subunit genes

J. Summary of GnRH action in GGH₃ cells

V. Studies of GnRH Action in Other Pituitary Cell Lines

A. RC-4B/C cells

B. LßT2 cells

VI. Future Directions

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				B. R. White, D. L. Duval, J. M. Mulvaney, M. S. Roberson, and C. M. Clay Homologous Regulation of the Gonadotropin-Releasing Hormone Receptor Gene Is Partially Mediated by Protein Kinase C Activation of an Activator Protein-1 Element Mol. Endocrinol., April 1, 1999; 13(4): 566 - 577. [Abstract] [Full Text]

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				S. Nelson, R. D. Horvat, J. Malvey, D. A. Roess, B. G. Barisas, and C. M. Clay Characterization of an Intrinsically Fluorescent Gonadotropin-Releasing Hormone Receptor and Effects of Ligand Binding on Receptor Lateral Diffusion Endocrinology, February 1, 1999; 140(2): 950 - 957. [Abstract] [Full Text]

				X. Lin and P. M. Conn Transcriptional Activation of Gonadotropin-Releasing Hormone (GnRH) Receptor Gene by GnRH: Involvement of Multiple Signal Transduction Pathways Endocrinology, January 1, 1999; 140(1): 358 - 364. [Abstract] [Full Text]

				F. Tejada, A. Cremades, M. Aviles, M. T. Castells, and R. Penafiel Hypokalemia alters sex hormone and gonadotropin levels: evidence that FSH may be required for luteinization Am J Physiol Endocrinol Metab, December 1, 1998; 275(6): E1037 - E1045. [Abstract] [Full Text] [PDF]

				X. Lin, J. A. Janovick, and P. M. Conn Mutations at the Consensus Phosphorylation Sites in the Third Intracellular Loop of the Rat Gonadotropin-Releasing Hormone Receptor: Effects on Receptor Ligand Binding and Signal Transduction Biol Reprod, December 1, 1998; 59(6): 1470 - 1476. [Abstract] [Full Text]

				B. D. Strahl, H.-J. Huang, J. Sebastian, B. R. Ghosh, and W. L. Miller Transcriptional Activation of the Ovine Follicle-Stimulating Hormone {beta}-Subunit Gene by Gonadotropin-Releasing Hormone: Involvement of Two Activating Protein-1-Binding Sites and Protein Kinase C Endocrinology, November 1, 1998; 139(11): 4455 - 4465. [Abstract] [Full Text] [PDF]

				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. [Abstract] [Full Text] [PDF]

				J. Weck, P. C. Fallest, L. K. Pitt, and M. A. Shupnik Differential Gonadotropin-Releasing Hormone Stimulation of Rat Luteinizing Hormone Subunit Gene Transcription by Calcium Influx and Mitogen-Activated Protein Kinase-Signaling Pathways Mol. Endocrinol., March 1, 1998; 12(3): 451 - 457. [Abstract] [Full Text]

				X. Lin, J. A. Janovick, S. Brothers, M. Blömenrohr, J. Bogerd, and P. M. Conn Addition of Catfish Gonadotropin-Releasing Hormone (GnRH) Receptor Intracellular Carboxyl-Terminal Tail to Rat GnRH Receptor Alters Receptor Expression and Regulation Mol. Endocrinol., February 1, 1998; 12(2): 161 - 171. [Abstract] [Full Text]

				M. Thibonnier, J. A. Preston, N. Dulin, P. L. Wilkins, L. N. Berti-Mattera, and R. Mattera The Human V3 Pituitary Vasopressin Receptor: Ligand Binding Profile and Density-Dependent Signaling Pathways Endocrinology, October 1, 1997; 138(10): 4109 - 4122. [Abstract] [Full Text] [PDF]

				E. J. Kaftan, T. Xu, R. F. Abercrombie, and B. Hille Mitochondria Shape Hormonally Induced Cytoplasmic Calcium Oscillations and Modulate Exocytosis J. Biol. Chem., August 11, 2000; 275(33): 25465 - 25470. [Abstract] [Full Text] [PDF]