The functions of estrogen receptors (ERs) in mouse ovary and genital tracts were investigated by generating null mutants for ERalpha (ERalphaKO), ERbeta (ERbetaKO) and both ERs (ERalphabetaKO). All ERalphaKO females are sterile, whereas ERbetaKO females are either infertile or exhibit variable degrees of subfertility. Mast cells present in adult ERalphaKO and ERalphabetaKO ovaries could participate in the generation of hemorrhagic cysts. Folliculogenesis proceeds normally up to the large antral stage in both ERalphaKO and ERbetaKO adults, whereas large antral follicles of ERalpha+/−ERbetaKO and ERalphabetaKO adults are markedly deficient in granulosa cells. Similarly, prematurely developed follicles found in prepubertal ERalphaKO ovaries appear normal, but their ERalphabetaKO counterparts display only few granulosa cell layers. Upon superovulation treatment, all prepubertal ERalphaKO females form numerous preovulatory follicles of which the vast majority do not ovulate. The same treatment fails to elicit the formation of preovulatory follicles in half of the ERbetaKO mice and in all ERalpha+/−/ERbetaKO mice. These and other results reveal a functional redundancy between ERalpha and ERbeta for ovarian folliculogenesis, and strongly suggest that (1) ERbeta plays an important role in mediating the stimulatory effects of estrogens on granulosa cell proliferation, (2) ERalpha is not required for follicle growth under wild type conditions, while it is indispensable for ovulation, and (3) ERalpha is also necessary for interstitial glandular cell development. Our data also indicate that ERbeta exerts some function in ERalphaKO uterus and vagina. ERalphabetaKO granulosa cells localized within degenerating follicles transform into cells displaying junctions that are unique to testicular Sertoli cells. From the distribution pattern of anti-Mullerian hormone (AMH) in ERalphabetaKO ovaries, it is unlikely that an elevated AMH level is the cause of Sertoli cell differentiation. Our results also show that cell proliferation in the prostate and urinary bladder of old ERbetaKO and ERalphabetaKO males is apparently normal.
REFERENCES
Adra
C. N.
, Boer
P. H.
, McBurney
M. W.
(1987
) Cloning and expression of the mouse pgk-1 gene and the nucleotide sequence of its promoter.
Gene
60
, 65
–74
Ahuja
H. S.
, Tenniswood
M.
, Lockshin
R.
, Zakeri
Z. F.
(1994
) Expression of clusterin in cell differentiation and cell death.
Biochem. Cell. Biol
72
, 523
–530
Al-Attar
L.
, Noël
K.
, Dutertre
M.
, Belville
C.
, Forest
M. G.
, Burgoyne
P. S.
, Josso
N.
, Rey
R.
(1997
) Hormonal and cellular regulation of Sertoli cell anti-Mullerian hormone production in the postnatal mouse.
J. Clin. Invest
100
, 1335
–1343
Aronow
B. J.
, Lund
S. D.
, Brown
T. L.
, Harmony
J. A. K.
, Witte
D. P.
(1993
) Apolipoprotein J expression at fluid-tissue interfaces: potential role in barrier cytoprotection.
Proc. Natl. Acad. Sci. USA
90
, 725
–729
Auffray
C.
, Rougeon
F.
(1980
) Purification of mouse immunoglobulin heavy-chain messenger RNAs from total myeloma tumor RNA.
Eur. J. Biochem
107
, 303
–314
Baarends
W. M.
, Uilenbroek
J. T. J.
, Kramer
P.
, Hoogerbrugge
J. W.
, van Leeuwen
E. C. M.
, Themmen
A. P. N.
, Grootegoed
J. A.
(1995
) Anti-mullerian hormone and anti-mullerian hormone type II receptor messenger ribonucleic acid expression in rat ovaries during postnatal development, the estrous cycle, and gonadotropin-induced follicle growth.
Endocrinology
136
, 4951
–4962
Behringer
R. R.
, Cate
R. L.
, Froelick
G. J.
, Palmiter
R. D.
, Brinster
R. L.
(1990
) Abnormal sexual development in transgenic mice chronically expressing mullerian inhibiting substance.
Nature
345
, 167
–170
Berry
M.
, Metzger
D.
, Chambon
P.
(1990
) Role of the two activating domains of the oestrogen receptor in the cell-type and promoter-context dependent agonistic activity of the anti-oestrogen 4-hydroxytamoxifen.
EMBO J
9
, 2811
–2818
Bonkhoff
H.
, Fixemer
T.
, Hunsicker
I.
, Remberger
K.
(1999
) Estrogen receptor expression in prostate cancer and premalignant prostatic lesions.
Am. J. Pathol
155
, 641
–647
Byers
M.
, Kuiper
G. G.
, Gustafsson
J. A.
, Park-Sarge
O. K.
(1997
) Estrogen receptor-mRNA expression in rat ovary: down-regulation by gonadotropins.
Mol. Endocrinol
11
, 172
–182
Couse
J. F.
, Curtis
S. W.
, Washburn
T. F.
, Lindzey
J.
, Golding
T. S.
, Lubahn
D. B.
, Smithies
O.
, Korach
K. S.
(1995
) Analysis of transcription and estrogen insensitivity in the female mouse after targeted disruption of the estrogen receptor gene.
Mol. Endocrinol
9
, 1441
–1454
Couse
J. F.
, Lindzey
J.
, Grandien
K.
, Gustafsson
J. A.
, Korach
K. S.
(1997
) Tissue distribution and quantitative analysis of estrogen receptor-(ER) and estrogen receptor-(ER) messenger ribonucleic acid in the wild-type and ER-knockout mouse.
Endocrinology
138
, 4613
–4621
Couse
J. F.
, Korach
K. S.
(1999
) Estrogen receptor null mice: what have we learned and where will they lead us?.
Endocr. Rev
20
, 358
–417
Couse
J. F.
, Bunch
D. O.
, Lindzey
J.
, Schomberg
D. W.
, Korach
K. S.
(1999
) Prevention of the polycystic ovarian phenotype and characterization of ovulatory capacity in the estrogen receptor-knockout mouse.
Endocrinology
140
, 5855
–5865
Couse
J. F.
, Curtis Hewitt
S.
, Bunch
D. O.
, Sar
M.
, Walker
V. R.
, Davis
B. J.
, Korach
K. S.
(1999
) Postnatal sex reversal of the ovaries in mice lacking estrogen receptorsand.
Science
286
, 2328
–2331
Dierich
A.
, Sairam
M. R.
, Monaco
L.
, Fimia
G. M.
, Gansmuller
A.
, LeMeur
M.
, Sassone-Corsi
P.
(1998
) Impairing follicle-stimulating hormone (FSH) signaling in vivo: targeted disruption of the FSH receptor leads to aberrant gametogenesis and hormonal imbalance.
Proc. Natl. Acad. Sci. USA
95
, 13612
–13617
Dupe
V.
, Davenne
M.
, Brocard
J.
, Dolle
P.
, Mark
M.
, Dierich
A.
, Chambon
P.
, Rijli
F. M.
(1997
) In vivo functional analysis of the Hoxa-1 3retinoic acid response element (3 RARE).
Development
124
, 399
–410
Eddy
E. M.
, Washburn
T. F.
, Bunch
D. O.
, Goulding
E. H.
, Gladen
B. C.
, Lubahn
D. B.
, Korach
K. S.
(1996
) Targeted disruption of the estrogen receptor gene in male mice causes alteration of spermatogenesis and infertility.
Endocrinology
137
, 4796
–4805
Fisher
C. R.
, Graves
K. H.
, Parlow
A. F.
, Simpson
E. R.
(1998
) Characterization of mice deficient in aromatase (ArKO) because of targeted disruption of the cyp19 gene.
Proc. Natl. Acad. Sci. USA
95
, 6965
–6970
Ghyselinck
N. B.
, Båvik
C.
, Sapin
V.
, Mark
M.
, Bonnier
D.
, Hindelang
C.
, Dierich
A.
, Nilsson
C. B.
, Håkansson
H.
, Sauvant
P.
, Azaïs-Braesco
V.
, Frasson
M.
, Picaud
S.
, Chambon
P.
(1999
) Cellular retinol-binding protein I is essential for vitamin A homeostasis.
EMBO J
18
, 4903
–4914
Green
S.
, Walter
P.
, Kumar
V.
, Krust
A.
, Bornert
J.M.
, Argos
P.
, Chambon
P.
(1986
) Human oestrogen receptor cDNA: sequence, expression and homology to v-erb-A.
Nature
320
, 134
–139
Habenicht
U. F.
, Tunn
U. W.
, Senge
T.
, Schröder
F. H.
, Schweikert
H. U.
, Bartsch
G.
, El Etreby
M. F.
(1993
) Management of benign prostatic hyperplasia with particular emphasis on aromatase inhibitors.
J. Steroid Biochem. Mol. Biol
44
, 557
–563
Hess
R. A.
, Bunick
D.
, Lee
K. H.
, Bahr
J.
, Taylor
J. A.
, Korach
K. S.
, Lubahn
D. B.
(1997
) A role for oestrogens in the male reproductive system.
Nature
390
, 509
–512
Hirobe
S.
, He
W. W.
, Lee
M. M.
, Donahoe
P. K.
(1992
) Mullerianinhibiting substance messenger ribonucleic acid expression in granulosa and Sertoli cells coincides with their mitotic activity.
Endocrinology
131
, 854
–862
Hunter
F.
, Leathem
J. H.
(1968
) Histamine concentrations in normal and cystic rat ovaries.
Endocrinology
82
, 171
–172
Jones
R. E.
, Duvall
D.
, Guillette
L. J.
, Lopez
Jr., Lopez
K. H.
(1994
) Number and state of rat ovarian mast cells after exogenous administration of luteinizing hormone.
Comp. Biochem. Physiol
108
, 555
–559
Kato
S.
, Endoh
H.
, Masuhiro
Y.
, Kitamoto
T.
, Uchiyama
S.
, Sasaki
H.
, Masushige
S.
, Gotoh
Y.
, Nishida
E.
, Kawashima
H.
, Metzger
D.
, Chambon
P.
(1995
) Activation of the estrogen receptor through phosphorylation by mitogen-activated protein kinase.
Science
270
, 1491
–1494
Krege
J. H.
, Hodgin
J. B.
, Couse
J. F.
, Enmark
E.
, Warner
M.
, Mahler
J. F.
, Sar
M.
, Korach
K. S.
, Gustafsson
J. A.
, Smithies
O.
(1998
) Generation and reproductive phenotypes of mice lacking estrogen receptor.
Proc. Natl. Acad. Sci. USA
95
, 15677
–15682
Krishna
A.
, Beesley
K.
, Terranova
P. F.
(1989
) Histamine, mast cells and ovarian function.
J. Endocrinol
120
, 363
–371
Krust
A.
, Green
S.
, Argos
P.
, Kumar
V.
, Walter
P.
, Bornert
J.M.
, Chambon
P.
(1986
) The chicken oestrogen receptor sequence: homology with v-erbA and the human oestrogen and glucocorticoid receptors.
EMBO J
5
, 891
–897
Kuhn
E. M.
, Therman
E.
(1988
) The behavior of heterochromatin in mouse and human nuclei.
Cancer Genet. Cytogenet
34
, 143
–351
Kuiper
G. G.
, Enmark
E.
, Pelto-Huikko
M.
, Nilsson
S.
, Gustafsson
J. A.
(1996
) Cloning of a novel receptor expressed in rat prostate and ovary.
Proc. Natl. Acad. Sci. USA
93
, 5925
–5930
Kuiper
G. G.
, Carlsson
B.
, Grandien
K.
, Enmark
E.
, Häggblad
J.
, Nilsson
S.
, Gustafsson
J. A.
(1997
) Comparison of the ligand binding specificity and transcript tissue distribution of estrogen receptorsand.
Endocrinology
138
, 863
–870
Ljungqvist
A.
, Feng
D. M.
, Hook
W.
, Shen
Z. X.
, Bowers
C.
, Folkers
K.
(1988
) Antide and related antagonists of luteinizing hormone release with long action and oral activity.
Proc. Natl. Acad. Sci USA
85
, 8236
–8240
Lu
B.
, Leygue
E.
, Dotzlaw
H.
, Murphy
L. J.
, Murphy
L. C.
, Watson
P. H.
(1998
) Estrogen receptor-mRNA variants in human and murine tissues.
Mol. Cell. Endocrinol
138
, 199
–203
Lubahn
D. B.
, Moyer
J. S.
, Golding
T. S.
, Couse
J. F.
, Korach
K. S.
, Smithies
O.
(1993
) Alteration of reproductive function but not prenatal sexual development after insertional disruption of the mouse estrogen receptor gene.
Proc. Natl. Acad. Sci. U S A
90
, 11162
–11166
Lufkin
T.
, Dierich
A.
, LeMeur
M.
, Mark
M.
, Chambon
P.
(1991
). Disruption of the Hox-1.6 homeobox gene results in defects in a region corresponding to its rostral domain of expression.
Cell
66
, 1105
–1119
Metzger
D.
, Clifford
J.
, Chiba
H.
, Chambon
P.
(1995
) Conditional site-specific recombination in mammalian cells using a ligand-dependent chimeric Cre recombinase.
Proc. Natl. Acad. Sci. USA
92
, 6991
–6995
Mosselman
S.
, Polman
J.
, Dijkema
R.
(1996
) ER: identification and characterization of a novel human estrogen receptor.
FEBS Lett
392
, 49
–53
Munsterberg
A.
, Lovell-Badge
R.
(1991
) Expression of the mouse anti-mullerian hormone gene suggests a role in both male and female sexual differentiation.
Development
113
, 613
–624
Muramatsu
M.
, Inoue
S.
(2000
) Estrogen receptors:how do they control reproductive and nonreproductive functions?.
Biochem. Biophys. Res. Commun
270
, 1
–10
Paech
K.
, Webb
P.
, Kuiper
G.G.
, Nilsson
S.
, Gustafsson
J.
, Kushner
P.J.
, Scanlan
T.S.
(1997
) Differential ligand activation of estrogen receptors ERand ER at AP1 sites.
Science
277
, 1508
–1510
Pelletier
R. M.
, Byers
S. W.
(1992
) The blood-testis barrier and Sertoli cell junctions: structural considerations.
Microsc. Res. Tech
20
, 3
–33
Risma
K. A.
, Clay
C. M.
, Nett
T. M.
, Wagner
T.
, Yun
J.
, Nilson
J. H.
(1995
) Targeted overexpression of luteinizing hormone in transgenic mice leads to infertility, polycystic ovaries, and ovarian tumors.
Proc. Natl. Acad. Sci. USA
92
, 1322
–1326
Risma
K. A.
, Hirshfield
A. N.
, Nilson
J. H.
(1997
) Elevated luteinizing hormone in prepubertal transgenic mice causes hyperandrogenemia, precocious puberty, and substantial ovarian pathology.
Endocrinology
138
, 3540
–3547
Robker
R. L.
, Richards
J. S.
(1998
) Hormone-induced proliferation and differentiation of granulosa cells: a coordinated balance of the cell cycle regulators cyclin D2 and p27Kip1.
Mol. Endocrinol
12
, 924
–940
Robker
R. L.
, Richards
J. S.
(1998
) Hormonal control of the cell cycle in ovarian cells: proliferation versus differentiation.
Biol. Reprod
59
, 476
–482
Rochette-Egly
C.
, Lutz
Y.
, Pfister
V.
, Heyberger
S.
, Scheuer
I.
, Chambon
P.
, Gaub
M. P.
(1994
) Detection of retinoid X receptors using specific monoclonal and polyclonal antibodies.
Biochem. Biophys. Res. Commun
204
, 525
–536
Rosenfeld
C. S.
, Ganjam
V. K.
, Taylor
J. A.
, Yuan
X.
, Stiehr
J. R.
, Hardy
M. P.
, Lubahn
D. B.
(1998
) Transcription and translation ofestrogen receptor-in the male reproductive tract of estrogen receptor- knock-out and wild-type mice.
Endocrinology
139
, 2982
–2987
Rosenfeld
C.S.
, Murray
A.A.
, Simmer
G.
, Hufford
M.G.
, Smith
M.F.
, Spears
N.
, Lubahn
D.B.
(2000
) Gonadotropin induction of ovulation and corpus luteum formation in young estrogen receptor-knockout mice.
Biol. Reprod
62
, 599
–605
Sar
M.
, Welsch
F.
(1999
) Differential expression of estrogen receptor-and estrogen receptor- in the rat ovary.
Endocrinology
140
, 963
–971
Saville
B.
, Wormke
M.
, Wang
F.
, Nguyen
T.
, Enmark
E.
, Kuiper
G.
, Gustafsson
J.-A.
, Safe
S.
(2000
) Ligand-, cell-, and estrogen receptor subtype (/)-dependent activation at GC-rich (Sp1) promoter elements.
J. Biol. Chem
275
, 5379
–5387
Schomberg
D. W.
, Couse
J. F.
, Mukherjee
A.
, Lubahn
D. B.
, Sar
M.
, Mayo
K. E.
, Korach
K. S.
(1999
) Targeted disruption of the estrogen receptor-gene in female mice: characterization of ovarian responses and phenotype in the adult.
Endocrinology
140
, 2733
–2744
Sicinski
P.
, Donaher
J. L.
, Geng
Y.
, Parker
S. B.
, Gardner
H.
, Park
M. Y.
, Robker
R. L.
, Richards
J. S.
, McGinnis
L. K.
, Biggers
J. D.
, Eppig
J. J.
, Bronson
R. T.
, Elledge
S. J.
, Weinberg
R. A.
(1996
) Cyclin D2 is an FSH-responsive gene involved in gonadal cell proliferation and oncogenesis.
Nature
384
, 470
–474
Swain
A.
, Lovell-Badge
R.
(1999
) Mammalian sex determination: a molecular drama.
Genes Dev
13
, 755
–767
Taketo-Hosotani
T.
, Merchant-Larios
H.
, Thau
R. B.
, Koide
S. S.
(1985
) Testicular cell differentiation in fetal mouse ovaries following transplantation into adult male mice.
J. Exp. Zool
236
, 229
–237
Tetsuka
M.
, Milne
M.
, Hillier
S. G.
(1998
) Expression of oestrogen receptor isoforms in relation to enzymes of oestrogen synthesis in rat ovary.
Mol. Cell. Endocrinol
141
, 29
–35
Tremblay
G. B.
, Tremblay
A.
, Copeland
N. G.
, Gilbert
D. J.
, Jenkins
N. A.
, Labrie
F.
, Giguere
V.
(1997
) Cloning, chromosomal localization, and functional analysis of the murine estrogen receptor.
Mol. Endocrinol
11
, 353
–365
Ueno
S.
, Takahashi
M.
, Manganaro
T. F.
, Ragin
R. C.
, Donahoe
P. K.
(1989
) Cellular localization of mullerian inhibiting substance in the developing rat ovary.
Endocrinology
124
, 1000
–1006
Vainio
S.
, Heikkilä
M.
, Kispert
A.
, Chin
N.
, McMahon
A. P.
(1999
) Female development in mammals is regulated by Wnt-4 signalling.
Nature
397
, 405
–409
Vigier
B.
, Watrin
F.
, Magre
S.
, Tran
D.
, Josso
N.
(1987
) Purified bovine AMH induces a characteristic freemartin effect in fetal rat prospective ovaries exposed to it in vitro.
Development
100
, 43
–55
Wang
H.
, Masironi
B.
, Eriksson
H.
, Sahlin
L.
(1999
) A comparative study of estrogen receptorsand in the rat uterus.
Biol. Reprod
61
, 955
–964
Weber
J. E.
, Turner
T. T.
, Tung
K. S.
, Russell
L. D.
(1988
) Effects of cytochalasin D on the integrity of the Sertoli cell (blood-testis) barrier.
Am. J. Anat
182
, 130
–147
White
R.
, Lees
J. A.
, Needham
M.
, Ham
J.
, Parker
M.
(1987
) Structural organization and expression of the mouse estrogen receptor.
Mol. Endocrinol
1
, 735
–744
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