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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