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Molecular Endocrinology 12 (7): 924-940
Copyright © 1998 by The Endocrine Society

Hormone-Induced Proliferation and Differentiation of Granulosa Cells: A Coordinated Balance of the Cell Cycle Regulators Cyclin D2 and p27Kip1

Rebecca L. Robker and JoAnne S. Richards

Department of Cell Biology Baylor College of Medicine Houston, Texas 77030

The proliferation and terminal differentiation of granulosa cells are critical for normal follicular growth, ovulation, and luteinization. Therefore, the in situ localization and hormonal regulation of cell cycle activators (cyclin D1, D2, and D3) and cell cycle inhibitors (p27Kip1 and p21Cip1) were analyzed in ovaries of mice and rats at defined stages of follicular growth and differentiation. Cyclin D2 mRNA was specifically localized to granulosa cells of growing follicles, while cyclin D1 and cyclin D3 were restricted to theca cells. In hypophysectomized (H) rats, cyclin D2 mRNA and protein were increased in granulosa cells by treatment with estradiol or FSH and were increased maximally by treatment with both hormones. In serum-free cultures of rat granulosa cells, cyclin D2 mRNA was rapidly elevated in response to FSH, forskolin, and estradiol, indicating that estradiol as well as cAMP can act directly and independently to increase cyclin D2 expression. The levels of p27Kip1 protein were not increased in response to estradiol or FSH. In contrast, when ovulatory doses of human CG (LH) were administered to hormonally primed H rats to stimulate luteinization, cyclin D2 mRNA and protein were rapidly decreased and undetectable within 4 h, specifically in granulosa cells of large follicles. Also in response to LH, the expression of the cell cycle inhibitor p27Kip1 was induced between 12 and 24 h (p21Cip1 was induced within 4 h) and remained elevated specifically in luteal tissue. A critical role for cyclin D2 in the hormone-dependent phase of follicular growth is illustrated by the ovarian follicles of cyclin D2-/- mice, which do not undergo rapid growth in response to hormones, but do express markers of FSH/LH action, cell cycle exit, and terminal differentiation. Collectively, these data indicate that FSH and estradiol regulate granulosa cell proliferation during the development of preovulatory follicles by increasing levels of cyclin D2 relative to p27Kip1 and that LH terminates follicular growth by down-regulating cyclin D2 concurrent with up-regulation of p27Kip1 and p21Cip1.




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J. Cottom, L. M. Salvador, E. T. Maizels, S. Reierstad, Y. Park, D. W. Carr, M. A. Davare, J. W. Hell, S. S. Palmer, P. Dent, et al.
Follicle-stimulating Hormone Activates Extracellular Signal-regulated Kinase but Not Extracellular Signal-regulated Kinase Kinase through a 100-kDa Phosphotyrosine Phosphatase
J. Biol. Chem., February 21, 2003; 278(9): 7167 - 7179.
[Abstract] [Full Text] [PDF]


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EndocrinologyHome page
N. A. Grieshaber, C. Ko, S. S. Grieshaber, I. Ji, and T. H. Ji
Follicle-Stimulating Hormone-Responsive Cytoskeletal Genes in Rat Granulosa Cells: Class I {beta}-Tubulin, Tropomyosin-4, and Kinesin Heavy Chain
Endocrinology, January 1, 2003; 144(1): 29 - 39.
[Abstract] [Full Text] [PDF]


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Genes Dev.Home page
M. A. Ciemerych, A. M. Kenney, E. Sicinska, I. Kalaszczynska, R. T. Bronson, D. H. Rowitch, H. Gardner, and P. Sicinski
Development of mice expressing a single D-type cyclin
Genes & Dev., December 15, 2002; 16(24): 3277 - 3289.
[Abstract] [Full Text] [PDF]


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EndocrinologyHome page
P. K. Pradeep, X. Li, H. Peegel, and K. M. J. Menon
Dihydrotestosterone Inhibits Granulosa Cell Proliferation by Decreasing the Cyclin D2 mRNA Expression and Cell Cycle Arrest at G1 Phase
Endocrinology, August 1, 2002; 143(8): 2930 - 2935.
[Abstract] [Full Text] [PDF]


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Biol. Reprod.Home page
N. Danilovich, D. Javeshghani, W. Xing, and M. R. Sairam
Endocrine Alterations and Signaling Changes Associated with Declining Ovarian Function and Advanced Biological Aging in Follicle-Stimulating Hormone Receptor Haploinsufficient Mice
Biol Reprod, August 1, 2002; 67(2): 370 - 378.
[Abstract] [Full Text] [PDF]


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Am. J. Physiol. Cell Physiol.Home page
J. E. Dumont, S. Dremier, I. Pirson, and C. Maenhaut
Cross signaling, cell specificity, and physiology
Am J Physiol Cell Physiol, July 1, 2002; 283(1): C2 - C28.
[Abstract] [Full Text] [PDF]


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Mol. Endocrinol.Home page
G. E. Owens, R. A. Keri, and J. H. Nilson
Ovulatory Surges of Human CG Prevent Hormone-Induced Granulosa Cell Tumor Formation Leading to the Identification of Tumor-Associated Changes in the Transcriptome
Mol. Endocrinol., June 1, 2002; 16(6): 1230 - 1242.
[Abstract] [Full Text] [PDF]


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Mol Hum ReprodHome page
S. Chu, S. Rushdi, E.T. Zumpe, P. Mamers, D.L. Healy, T. Jobling, H.G. Burger, and P.J. Fuller
FSH-regulated gene expression profiles in ovarian tumours and normal ovaries
Mol. Hum. Reprod., May 1, 2002; 8(5): 426 - 433.
[Abstract] [Full Text] [PDF]


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Mol. Endocrinol.Home page
J. S. Richards, S. C. Sharma, A. E. Falender, and Y. H. Lo
Expression of FKHR, FKHRL1, and AFX Genes in the Rodent Ovary: Evidence for Regulation by IGF-I, Estrogen, and the Gonadotropins
Mol. Endocrinol., March 1, 2002; 16(3): 580 - 599.
[Abstract] [Full Text] [PDF]


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EndocrinologyHome page
M. Hsieh, M. A. Johnson, N. M. Greenberg, and J. S. Richards
Regulated Expression of Wnts and Frizzleds at Specific Stages of Follicular Development in the Rodent Ovary
Endocrinology, March 1, 2002; 143(3): 898 - 908.
[Abstract] [Full Text] [PDF]


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EndocrinologyHome page
D. S. Moons, S. Jirawatnotai, T. Tsutsui, R. Franks, A. F. Parlow, D. B. Hales, G. Gibori, A. T. Fazleabas, and H. Kiyokawa
Intact Follicular Maturation and Defective Luteal Function in Mice Deficient for Cyclin- Dependent Kinase-4
Endocrinology, February 1, 2002; 143(2): 647 - 654.
[Abstract] [Full Text] [PDF]


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Reproductive SciencesHome page
D. Choi, S. Yoon, E. Lee, S. Hwang, S. Song, J. Kim, B.-K. Yoon, and J.-H. Lee
Characterization of Cyclin D2 Expression in Human Endometrium
Reproductive Sciences, January 1, 2002; 9(1): 41 - 46.
[Abstract] [PDF]


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Recent Prog Horm ResHome page
J. S. Richards, D. L. Russell, S. Ochsner, M. Hsieh, K. H. Doyle, A. E. Falender, Y. K. Lo, and S. C. Sharma
Novel Signaling Pathways That Control Ovarian Follicular Development, Ovulation, and Luteinization
Recent Prog. Horm. Res., January 1, 2002; 57(1): 195 - 220.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
L. M. Salvador, Y. Park, J. Cottom, E. T. Maizels, J. C. R. Jones, R. V. Schillace, D. W. Carr, P. Cheung, C. D. Allis, J. L. Jameson, et al.
Follicle-stimulating Hormone Stimulates Protein Kinase A-mediated Histone H3 Phosphorylation and Acetylation Leading to Select Gene Activation in Ovarian Granulosa Cells
J. Biol. Chem., October 19, 2001; 276(43): 40146 - 40155.
[Abstract] [Full Text] [PDF]


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EndocrinologyHome page
T. El-Hefnawy and A. J. Zeleznik
Synergism Between FSH and Activin in the Regulation of Proliferating Cell Nuclear Antigen (PCNA) and Cyclin D2 Expression in Rat Granulosa Cells
Endocrinology, October 1, 2001; 142(10): 4357 - 4362.
[Abstract] [Full Text] [PDF]


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Biol. Reprod.Home page
C. L. Chaffin, K. M. Schwinof, and R. L. Stouffer
Gonadotropin and Steroid Control of Granulosa Cell Proliferation During the Periovulatory Interval in Rhesus Monkeys
Biol Reprod, September 1, 2001; 65(3): 755 - 762.
[Abstract] [Full Text] [PDF]


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EndocrinologyHome page
K. H. Burns, C. Yan, T. R. Kumar, and M. M. Matzuk
Analysis of Ovarian Gene Expression in Follicle-Stimulating Hormone {beta} Knockout Mice
Endocrinology, July 1, 2001; 142(7): 2742 - 2751.
[Abstract] [Full Text] [PDF]


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Endocr. Rev.Home page
S. F. Palter, A. B. Tavares, A. Hourvitz, J. D. Veldhuis, and E. Y. Adashi
Are Estrogens of Import to Primate/Human Ovarian Folliculogenesis?
Endocr. Rev., June 1, 2001; 22(3): 389 - 424.
[Abstract] [Full Text] [PDF]


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EndocrinologyHome page
J. S. Richards
Perspective: The Ovarian Follicle--A Perspective in 2001
Endocrinology, June 1, 2001; 142(6): 2184 - 2193.
[Full Text] [PDF]


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EndocrinologyHome page
K. F. Roby
Alterations in Follicle Development, Steroidogenesis, and Gonadotropin Receptor Binding in a Model of Ovulatory Blockade
Endocrinology, June 1, 2001; 142(6): 2328 - 2335.
[Abstract] [Full Text] [PDF]


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JCBHome page
R. S. Muraoka, A. E.G. Lenferink, J. Simpson, D. M. Brantley, L. R. Roebuck, F. M. Yakes, and C. L. Arteaga
Cyclin-Dependent Kinase Inhibitor P27Kip1 Is Required for Mouse Mammary Gland Morphogenesis and Function
J. Cell Biol., May 28, 2001; 153(5): 917 - 932.
[Abstract] [Full Text] [PDF]


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Mol. Endocrinol.Home page
D. Waltregny, I. Leav, S. Signoretti, P. Soung, D. Lin, F. Merk, J. Y. Adams, N. Bhattacharya, N. Cirenei, and M. Loda
Androgen-Driven Prostate Epithelial Cell Proliferation and Differentiation in Vivo Involve the Regulation of p27
Mol. Endocrinol., May 1, 2001; 15(5): 765 - 782.
[Abstract] [Full Text]


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Mol. Endocrinol.Home page
J. S. Richards
New Signaling Pathways for Hormones and Cyclic Adenosine 3',5'-Monophosphate Action in Endocrine Cells
Mol. Endocrinol., February 1, 2001; 15(2): 209 - 218.
[Abstract] [Full Text]


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Reproductive SciencesHome page
J. S. Richards
Graafian Follicle Function and Luteinization in Nonprimates
Reproductive Sciences, January 1, 2001; 8(1_suppl): S21 - S23.
[Abstract] [PDF]


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EndocrinologyHome page
S. K. Roy and L. Albee
Requirement for Follicle-Stimulating Hormone Action in the Formation of Primordial Follicles during Perinatal Ovarian Development in the Hamster
Endocrinology, December 1, 2000; 141(12): 4449 - 4456.
[Abstract] [Full Text] [PDF]


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Biol. Reprod.Home page
T. L. Beumer, H. L. Roepers-Gajadien, I. S. Gademan, H. B. Kal, and D. G. de Rooij
Involvement of the D-Type Cyclins in Germ Cell Proliferation and Differentiation in the Mouse
Biol Reprod, December 1, 2000; 63(6): 1893 - 1898.
[Abstract] [Full Text]


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EndocrinologyHome page
N. A. Grieshaber, S. Boitano, I. Ji, J. P. Mather, and T. H. Ji
Differentiation of Granulosa Cell Line: Follicle-Stimulating Hormone Induces Formation of Lamellipodia and Filopodia via the Adenylyl Cyclase/Cyclic Adenosine Monophosphate Signal
Endocrinology, September 1, 2000; 141(9): 3461 - 3470.
[Abstract] [Full Text] [PDF]


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Mol. Endocrinol.Home page
I. J. Gonzalez-Robayna, A. E. Falender, S. Ochsner, G. L. Firestone, and J. S. Richards
Follicle-Stimulating Hormone (FSH) Stimulates Phosphorylation and Activation of Protein Kinase B (PKB/Akt) and Serum and Glucocorticoid-Induced Kinase (Sgk): Evidence for A Kinase-Independent Signaling by FSH in Granulosa Cells
Mol. Endocrinol., August 1, 2000; 14(8): 1283 - 1300.
[Abstract] [Full Text]


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EndocrinologyHome page
L. L. Espey, T. Ujioka, D. L. Russell, M. Skelsey, B. Vladu, R. L. Robker, H. Okamura, and J. S. Richards
Induction of Early Growth Response Protein-1 Gene Expression in the Rat Ovary in Response to an Ovulatory Dose of Human Chorionic Gonadotropin
Endocrinology, July 1, 2000; 141(7): 2385 - 2391.
[Abstract] [Full Text] [PDF]


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EndocrinologyHome page
K. L. Britt, A. E. Drummond, V. A. Cox, M. Dyson, N. G. Wreford, M. E. E. Jones, E. R. Simpson, and J. K. Findlay
An Age-Related Ovarian Phenotype in Mice with Targeted Disruption of the Cyp 19 (Aromatase) Gene
Endocrinology, July 1, 2000; 141(7): 2614 - 2623.
[Abstract] [Full Text] [PDF]


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Biol. Reprod.Home page
B. D. Murphy
Models of Luteinization
Biol Reprod, July 1, 2000; 63(1): 2 - 11.
[Abstract] [Full Text]


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Biol. Reprod.Home page
A. Hampl, J. Pacherník, and P. Dvorák
Levels and Interactions of p27, Cyclin D3, and CDK4 During the Formation and Maintenance of the Corpus Luteum in Mice
Biol Reprod, May 1, 2000; 62(5): 1393 - 1401.
[Abstract] [Full Text]


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DevelopmentHome page
S Dupont, A Krust, A Gansmuller, A Dierich, P Chambon, and M Mark
Effect of single and compound knockouts of estrogen receptors alpha (ERalpha) and beta (ERbeta) on mouse reproductive phenotypes
Development, January 10, 2000; 127(19): 4277 - 4291.
[Abstract] [PDF]


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EndocrinologyHome page
T. N. Alliston, I. J. Gonzalez-Robayna, P. Buse, G. L. Firestone, and J. S. Richards
Expression and Localization of Serum/Glucocorticoid-Induced Kinase in the Rat Ovary: Relation to Follicular Growth and Differentiation
Endocrinology, January 1, 2000; 141(1): 385 - 395.
[Abstract] [Full Text] [PDF]




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