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Key Words. estrogen receptor, reproductive tract, transgenic mice, for which no ligands have been identied [1]. A second
estrogen action ER gene was cloned from prostate tissue in 1996 [2], and
thus, there are two ER molecules: the original ER ``a'',
and the recently discovered ER ``b''. By comparing the
Introduction ER sequences it is apparent that both share a general
domain structure common to ligand modulated nuclear
The biological effects of estrogen (E) in mammalian target transcription factors [3]. The current understanding of
tissues are important for numerous physiological pro- ER mechanisms of action can be summarized with
cesses. E is known to induce responses in the reproductive reference to the overall structure of the ER molecules
tract, mammary tissue and pituitary but also affects non- (Fig. 1). The functions of some regions of the ER
reproductive processes such as bone formation and molecules have been dened using deletion and mutation
cardiovascular health. Since the initial identication and as well as structural analysis. The best characterized
isolation of the estrogen receptor (ER) molecule decades functions are a zinc-nger containing domain (C domain,
ago, the mechanism of estrogen (E) action in cells has been Fig. 1) that binds with high afnity and specicity to
intensely studied. At the core, our understanding of the EREs in target genes, and a ligand-binding domain
basic mechanism has remained the same, however, the (LBD, domain E), which binds E as well as other
details continue to be dened and many new aspects estrogenic ligands. The ERE is a 13 base pair inverted
remain to be discovered. The dogma of high afnity E repeat sequence (GGTCAnnnTGACC), and in vitro DNA
binding and modulation of transcription via high afnity binding studies have indicated that the ER binds as a
estrogen responsive elements (EREs) in target genes still dimer [4], with one ER molecule contacting each 5 base
remains the basis of E action in target tissues. However, pair inverted repeat [5]. Although the DNA binding is a
this model has become much more complex with the dimerization stimulus, sequences in the LBD are also
understanding that many other factors cooperate and involved in dimerization [4], and crystallized truncated
interact to modulate transcription. In addition there are ER containing only ligand binding domain is clearly
``twists'' in the ``plot'', in alternatives to E activation of shown to be a dimer in the presence of ligand [6].
ER as well as ER's ability to modulate transcription of In addition, regions in the amino terminus (AF-1) and
genes that lack EREs. Advances in technology and within the LBD (AF-2) are involved in ligand-indepen-
methods available for molecular, structural, biochemical, dent (AF-1) and ligand-dependent (AF-2) transcriptional
genetic and physiological analysis of ER function have activation, as deletion of or introduction of mutations in
allowed for the denition of many of the details of ER these regions result in a diminished ability to induce E
function in in vitro model systems and the expansion of responsive genes [7,8]. The mechanism by which
these models into genetically modied mouse models. transcription is mediated by ER is thought to be through
This chapter will review the current understanding of the interaction of AFs with ``transcriptional machinery'',
mechanism of ER function and then describe insights which is a general term referring to the complex of
gained from use of transgenic mouse models. molecules that assembles and ultimately results in
synthesis of mRNA [9]. Much is now known about
RNA polymerase II and the enzymes and factors that
Estrogen Receptors: Structure and allow transcription. Transcriptional co-regulators are
Mechanism of Action molecules that mediate the interaction between ERs
and the transcriptional machinery. Many co-regulators
ERs are members of a family of nuclear transcription
factors including receptors for sex steroids, thyroid Address correspondence to: Kenneth S. Korach.
hormone, retinoids as well as many ``orphan'' receptors, E-mail: korach@niehs.nih.gov
193
194 Hewitt and Korach
Table 1. The Phenotypes reported in mice with disrupted ERa, ERb, or both ERa and ERb
Tissue a b ab
receptor pathways and nuclear receptors has been the bERKO males are fertile and the bERKO females are
proposed (reviewed in Yee and Lee [25] and Cenni and subfertile [29,30].
Picard [26]). In support of this hypothesis, the estrogen
receptor a knockout (aERKO) mouse, which lacks ERa,
has been used to show that this receptor is essential for Role of ER in Female Fertility
EGF-induced DNA synthesis in uterine epithelium [27].
Further complicating these alternative modes of ER The most signicant phenotype in the aERKO and
activity, EGF and IGF-1 can stimulate a reporter gene bERKO females is complete infertility in the aERKO and
driven by GC rich promoter element from the cathepsin decreased fertility of the bERKO as a result of failure of
D gene [18]. several components necessary for successful reproduc-
tion. Normally, gonadotropin production by the pituitary
is regulated by a negative feedback loop in which rising
Animal Models for the Study of ER E levels down-regulate transcription of the LHb gene,
and thus, withdrawal of E by ovariectomy causes an
The structure and mechanisms of ER-mediated regula- increase in serum LH. The aERKO female also has
tion of various target genes have been extensively elevated LH transcript in the pituitary as well as elevated
characterized using isolated components or in vitro cell serum LH [31]. In contrast, LH levels are normal in the
models. The ability to produce transgenic mice with bERKO (unpublished), indicating that the negative
disruption of the ER genes has allowed study of the feedback is mediated by ERa rather than ERb.
physiological processes that require ERs in live animals Immunohistochemical analysis of the expression of
and provided insight into differing physiological roles of ERa and ERb protein in a normal ovary indicate that,
ERa and ERb. Table 1 summarizes many of the although both ERs are present, their distribution differs,
phenotypes reported in mice with disrupted ERa with ERb predominantly in the granulosa cells of the
(aERKO), ERb (bERKO) or both ERa and ERb follicles and ERa localized in the thecal and interstitial
(abERKO) (reviewed in Couse and Korach [28]). The regions of the ovary (Fig. 3). Therefore, one might expect
most signicant effects, as might be expected, are seen in the a or b ERKO mouse ovaries to exhibit different
reproductive systems of these animals. Both male and phenotypes. The aERKO ovaries develop hemorrhagic
female aERKO and abERKO mice are infertile, whereas cysts [29], and lack mature follicles and corpora lutea,
196 Hewitt and Korach
Fig. 4. Ovarian histology, reproductive tracts and mammary gland whole mounts from WT, aERKO, bERKO and abERKO tissues. Arrows indicate
rudimentary ductal tree in mammay glands.
animals [43]. When germ cells from aERKO males were follicles, containing an oocyte, a normal complement of
transplanted into testis of germ-cell depleted WT recipi- granulosa cells, and surrounding theca. However, there is
ents, normal offspring were produced by the recipients also a marked presence of abnormal follicles that, when
[44,45]. This experimental observation combined with the viewed in a 2-dimensional section, appear similar to
ability of ERa-disrupted sperm from ERa heterozygous seminiferous tubules. These follicles most often lack an
(ERa + ) males to produce offspring indicate that ERa oocyte and granulosa cells, but rather possess Sertoli-like
is not needed for sperm function, but is required in the cells, located along the basement membrane and
male reproductive tract to allow maturation of sperm. exhibiting the characteristic cytosolic extensions and
Although the lack of sperm alone results in male tripartite nucleoli. Further analysis indicates the presence
infertility, it should be noted that the aERKO and of what may be intermediate structures, i.e., follicles
abERKO males also exhibit decient mating behaviors containing a degenerating oocyte, surrounded by both
[46,47], while the bERKO mating behaviors are normal granulosa cells and Sertoli-like cells. Because this
[41], indicating the importance of ERa in normal male phenotype emerges progressively with age, it is currently
mating behaviors. thought that these tubular structures represent ``ghosts''
of follicles in which the germ cell has died and a
population of remaining granulosa cells is ``trans-
Phenotypes in the abERKO differentiated'' into Sertoli-like cells.
Fig. 6. Details of follicles from abERKO ovary. All three representative follicles are from the same ovary. The top panel shows a normal
developing follicle, with higher power detail to right showing granulosa cells. The bottom panel shows a seminiferous tubule-like structure with
higher power detail to right. Note the Sertoli-like cells along the basement membrane. The middle panel shows an intermediate follicle with
characteristics of both.
Estrogen Receptors 199
women indicates a possible role for E in bone and cardio- 4. Glass CK. Differential recognition of target genes by nuclear
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The shorter femurs are associated with lower serum IGF- view of the oestrogen receptor. J Steroid Biochem Mol Biol
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1 levels. aERKO femurs are also of smaller diameter
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9. Edwards DP. The role of coactivators and corepressors in the
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14. Watanabe M, Yanagisawa J, Kitagawa H, Takeyama K, Ogawa S,
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