| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
ARTICLES |
Departments of Cell and Structural Biology, and Molecular and Integrative Physiology, University of Illinois (T.R.E., B.S.K.), Urbana, Illinois 61801; the Department of Biology, University of California (W.L.K.), La Jolla, California 92093; the Department of Medicine, West Virginia University Medical Center (E.J.W.), Morgantown, West Virginia 26506; and the Department of Veterans Affairs Medical Center (E.J.W.), Clarksburg, West Virginia 26301
Address all correspondence and requests for reprints to: Dr. Benita S. Katzenellenbogen, Department of Molecular and Integrative Physiology, University of Illinois, 524 Burrill Hall, 407 South Goodwin Avenue, Urbana, Illinois 61801-3704. E-mail: katzenel{at}uiuc.edu
 |
Abstract |
|---|
 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
6-fold) by 1 h, and is not blocked
by cycloheximide, indicating that it is a primary response. Stimulation
is selective for estrogen ligands, with no stimulation by other classes
of steroid hormones, and stimulation by estrogen is suppressed by the
antiestrogens tamoxifen and ICI 182,780. Induction is shown to require
an active ER through several approaches, including the use of
ER-negative breast cancer cells containing a stably integrated ER.
NHE-RF protein levels, monitored using antibodies specific for this
protein, increase after estrogen and reach maximal levels at 24–48 h.
Interestingly, NHE-RF is a PDZ domain-containing protein that is
enriched in polarized epithelia, where it is known to be localized in
microvilli. Among various human tissues we have examined, we found that
NHE-RF is expressed at a fairly high level in mammary tissue. NHE-RF
regulates protein kinase A inhibition of the
Na+-H+ exchanger and may serve as a scaffold
adaptor protein that contributes to the specificity of signal
transduction events. Our findings suggest that the early, known effects
of estrogen on cell cytoarchitecture (e.g. increasing
microvilli on breast cancer cells) and on some cell signaling pathways
(e.g. those involving cAMP) may involve rapid
estrogen-mediated changes in the production of NHE-RF. |
Introduction |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
NHE-RF was first identified in kidney as a protein factor that was required for cAMP regulation of the activity of sodium/hydrogen exchanger type 3 (5). Intriguingly, very recent work has shown this protein to be identical to the protein named EBP50 (6), a PDZ domain-containing cytoskeletal protein associated with microfibrils and the cell cytoskeleton that is highly concentrated in cell microvilli. It is believed to play an important role in cell signaling associated with changes in cell cytoarchitecture. However, nothing is currently known about its regulation by hormones or other factors.
We show here that NHE-RF is dramatically up-regulated by estrogens, with substantial changes in its RNA level being detectable as early as 1 h after hormone treatment. As estrogenic hormones are known to have marked effects on cell cytoarchitecture and the development of microvilli (2) and concomitant early effects on cell signaling (3, 7, 8, 9), these observations of NHE-RF regulation by estrogen raise intriguing questions regarding the links between estrogens and cytoskeletal, cell signaling, and cAMP-regulated events.
|
Materials and Methods |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
-dihydrotestosterone, and
cycloheximide were purchased from Sigma Chemical Co. (St.
Louis, MO). The antiestrogens ICI182,780 (ICI) and
trans-hydroxytamoxifen (TOT) were provided by Alan Wakeling
and Zeneca Pharmaceuticals (Macclesfield, UK). The
synthetic estrogen P1496 was obtained from IMC Corp. (Terre Haute, IN).
The progestin R5020 was purchased from Roussell-UCLAF (Romainville,
France).
Cell culture
MCF-7 human breast cancer cells, originally acquired from the
Michigan Cancer Foundation, were maintained in growth medium,
which was MEM plus phenol red supplemented with 5% heat-inactivated
FCS, 10-12 M estradiol (E2), and
10 mM HEPES. Before the experiments, cells were depleted of
estrogen by growth for 1 week in the same growth medium without
E2. MCF-7 cells were then grown in MEM plus phenol red
supplemented with 5% charcoal-dextran-stripped FCS for 2 days, after
which the cells were maintained in improved MEM (IMEM) minus phenol red
plus 5% charcoal-dextran-stripped FCS for 8 days before use in
experiments. All media included penicillin (100 U/ml), streptomycin
(100 µg/ml), and gentamicin (25 µg/ml; Life Technologies).
MDA-MB-231 human breast cancer cells were maintained in DMEM-Ham’s F-12 nutrient mixture with 10 mM HEPES, 10% FCS, penicillin (100 U/ml), streptomycin (100 µg/ml), gentamicin (25 µg/ml), bovine insulin (6 ng/ml), hydrocortisone (3.75 ng/ml), and glutathione (16 µg/ml).
The 231/ER+ cells were generated from the MDA-MB-231 cells
by stable transfection of an expression vector for human ER
(pCMV5-hER) (10, 11) and a separate expression vector containing the
neor gene to confer resistance to geneticin.
This clone (no. 20) was selected as an isolated colony after drug
selection and was shown to express ER at 550 ± 66 fmol/mg total
cellular protein. These cells were maintained in the same medium as
parental cells with perpetual drug selection [600 µg geneticin
(active)/ml culture medium]. Before experiments, cells were grown
without geneticin for several weeks. Cells used in all experiments were
80–95% confluent.
Isolation of RNA
Total RNA used for differential display was isolated using the
RNA extraction kit from Pharmacia Biotech (Piscataway, NJ)
following the manufacturer’s instructions. Total RNA used for Northern
transfer was isolated using the RNA Stat-60 kit (Tel-Test,
Inc., Friendswood, TX).
Differential display
Differential display PCR (ddPCR) was performed using the
RNAimage kit from GenHunter (Nashville, TN) as previously described
(12). DNA-free RNA was obtained by treatment of total RNA with
deoxyribonuclease (Promega Corp., Madison, WI) in the
presence of ribonuclease inhibitor (Promega Corp.) for 30
min at 37 C. After phenol/chloroform extraction and ethanol
precipitation, three RT reactions were performed for each RNA sample
using 0.2 µg DNA-free total RNA in 1 x RT buffer [25
mM Tris-Cl (pH 8.3), 37.6 mM KCl, 1.5
mM MgCl2, and 5 mM
dithiothreitol]; 20 µM each of deoxy (d)-ATP, dCTP,
dGTP, and dTTP; and 0.2 µM of one HindIII
restriction site-containing one-base-anchored oligo(deoxythymidine)
primer [H-T11A (5'-AAGCTTTTTTTT-TTTA-3'), H-T11C
(5'-AAGCTTTTTTTTTTTC-3'), or H-T11G (5'-AAGCTTTTTTTTTTTG-3')]. After
the solution was heated to 65 C for 5 min and cooled to 37 C, 100 U
Moloney murine leukemia virus reverse transcriptase were added for
1 h. PCR reactions were performed in mixtures containing 0.1 vol
RT mixture, 1 x PCR buffer [10 mM Tris-Cl (pH 8.4),
50 mM KCl, 1.5 mM MgCl2, and
0.001% gelatin]; 2 µM each of dATP, dCTP, dGTP, and
dTTP; 0.2 µM of HindIII restriction
site-containing arbitrary 13-mer oligonucleotide [H-AP1
(5'-AAGCTTGATTGCC-3'), H-AP2 (5'-AAGCTTCGACTGT-3'), H-AP3
(5'-AAGCTTTGGTCAG-3'), H-AP4 (5'-AAGCTTCTCAACG-3'), H-AP5
(5'-AAGCT-TAGTAGGC’3'), H-AP6 (AAGCTTGCACCAT-3'), H-AP7
(5'-AAGCTTAACGAGG-3'), or H-AP8 (5'-AAGCTTTTACCGC-3')]; 0.2
µM of the corresponding anchored oligo(deoxythymidine)
primer; 0.1 µCi [35S]dATP; and 1 U AmpliTaq DNA
polymerase (Perkin Elmer/Cetus, Norwalk, CT). Light
mineral oil was overlaid, and the PCR reactions were performed at 94 C
for 30 sec, followed by 40 cycles of 94 C for 30 sec, 40 C for 2 min,
and 72 C for 30 sec, followed by 72 C for 5 min. Stop buffer [95%
formamide, 10 mM EDTA (pH 8.0), 0.09% xylene cyanol, and
0.09% bromophenol blue] was added to each sample and heated at 80 C
for 2 min before loading on a 6% denaturing polyacrylamide sequencing
gel. After electrophoresis, the gels were exposed to Kodak XAR-5 film
(Eastman Kodak Co., Rochester, NY) for 48 h. Any band
differentially expressed was identified, and the PCR was repeated to
confirm the findings.
The differentially expressed complementary DNA (cDNA) species was recovered from the dried sequencing gel by elution in water and reamplified using the same primer set and PCR conditions as those used in the messenger RNA (mRNA) display, except that the dNTP concentration was 20 µM instead of 2 µM, and no isotope was added. Reamplified cDNA was run on a 1.2% agarose gel and purified using the QIAEX kit from QIAGEN (Chatsworth, CA). Reamplified cDNA was subsequently cloned into the PCRII vector using the TA cloning system from Invitrogen (San Diego, CA) and sequenced using the Sequenase kit (U.S. Biochemical Corp., Cleveland, OH).
Northern blot analysis
Twenty micrograms of total RNA were separated by electrophoresis
in a 1% agarose-7% formaldehyde gel and transferred to a GeneScreen
Plus nylon membrane (DuPont NEN, Boston, MA) by capillary
action in 10 x SSC (standard saline citrate). Prehybridization
was performed in Rapidhybe buffer (Amersham, Arlington
Heights, IL) at 65 C. Hybridization was performed in Rapidhybe buffer
at 65 C for 18 h with random primer-labeled cDNA, prepared
using the Rediprime DNA labeling system from Amersham.
After hybridization, the filters were washed three times for 3 min each
time at 25 C in 1 x SSC-0.1% SDS, followed by a 30-min wash in
0.5 x SSC-0.1% SDS at 65 C, followed by a high stringency wash
for 30 min at 65 C in 0.1 x SSC-0.1% SDS. The filters were
exposed to x-ray film at -80 C overnight. Message size was determined
by comparison with the migration of an RNA ladder (0.24–9.6 kb;
Life Technologies). Equalization of loading was controlled
by cohybridization with a cDNA probe for 36B4 that is not influenced by
the addition of steroid hormones or their antagonists (13). Signal
intensity was quantified by phosphorimager analysis and standardized to
the 36B4 signal.
A human tissue RNA blot containing equal amounts of tissue RNA per dot spot (CLONTECH Laboratories, Inc., Palo Alto, CA) was hybridized with NHE-RF cDNA, and NHE-RF was quantified by phosphorimager analysis.
Western blot analysis
After treatment with hormone or vehicle control, MCF-7 and
231/ER+ cells were harvested in HBSS (Life Technologies, Inc.)/trypsin-EDTA solution (Sigma Chemical Co.), centrifuged at 300 x g for 5 min, and
resuspended in 20 mM Tris (pH 7.4), 0.5 M NaCl,
1.0 mM dithiothreitol, 10% glycerol (vol/vol), 50 µg/ml
leupeptin, 50 µg/ml aprotinin, 2.5 µg/ml pepstatin, and 0.2
mM phenylmethysulfonylfluoride. Whole cell extracts were
obtained by subjecting cells to three rounds of freezing on dry ice and
thawing on wet ice, followed by centrifugation at 15,000 x
g to remove cell debris. Equal amounts of total protein were
loaded on a 10% SDS-polyacrylamide gel. Electrophoresis and Western
blotting were performed according to standard methods (14), and a 14.3-
to 220-kDa protein molecular mass marker set (Amersham)
was used for size comparison. Nitrocellulose blots were probed with the
NHE-RF primary antibody (polyclonal antibody made to full-length rabbit
NHE-RF) (15), then incubated with secondary antibody in blocking
solution and detected using an enhanced chemical luminescence system
(Amersham).
|
Results |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
). The differential display
product showed 60% sequence identity to the 3'-untranslated region of
the rabbit protein cofactor Na+/H+ exchanger
regulatory factor (NHE-RF; GenBank accession no. U19815). The
reamplified ddPCR product was labeled and used to probe a Northern
blot, which confirmed that the expression of NHE-RF (2.0 kb) is 4–5
times higher in the estradiol-treated cells (Fig. 1B). The human
expressed sequence tag (EST) database was found to contain a cDNA clone
with 99% sequence identity with the ddPCR fragment and which was
substantially longer than the isolated fragment, although not full
length (991 bp from the 3'-region of the gene). Probing the Northern
blot with the labeled EST clone gave results identical to those
obtained with the ddPCR probe; thus, it was used for the remaining
experiments.
|
). In contrast, when the same
cells contained a stably integrated ER expression construct (subline
231/ER+), estradiol elevated the NHE-RF mRNA level 5- to
6-fold over that in the untreated cells (Fig. 2). Hormone binding,
Western blot, and trans-activation studies with these
231/ER+ cells (not presented) demonstrated that the ER
was present (550 ± 66 fmol/mg protein) and retained function
within the stably transfected cells. Similar trends in E2
stimulation of NHE-RF were seen in another ER-positive breast cancer
cell line, ZR-75–1, whereas no E2 stimulation was seen in
a different ER-negative breast cancer cell line, MDA-MB-435S (data not
shown).
|
). There was a rapid and near-maximal
increase in the mRNA level as early as 1 h after treatment, as
indicated by Northern blot analysis. Levels peaked between 4–8 h and
then declined thereafter, but still remained elevated above the basal
value even at 24 h. Cells were treated with varying concentrations
of E2, and Northern blot analysis was performed on the
isolated RNA (Fig. 3B). Results show that the expression of NHE-RF mRNA
is dependent on the concentration of E2, with maximal
stimulation at 10-8 M E2.
|
-dihydrotestosterone
and testosterone, and the progestin R5020. Northern analysis of total
RNA showed that both estrogens increased NHE-RF mRNA levels, but that
the other hormones did not (Fig. 4A). The
antiestrogens ICI 182,780 and trans-hydroxytamoxifen were
also examined to determine whether they blocked the estrogen induction
of NHE-RF mRNA. Indeed, neither ligand stimulated mRNA expression by
itself, and cotreatment of the cells with E2 and the
antiestrogen reduced mRNA expression to that in untreated cells (Fig. 4B).
|
). In the 231/ER+ cells,
which have a much lower basal level of this protein, there was a steady
increase in the amount of NHE-RF protein, with 5-fold elevation over
the basal level observed after 48 h of estradiol treatment. In
addition, cotreatment with E2 and ICI 182,780 reduced
NHE-RF protein expression to basal levels (data not shown).
|
, A
and B). Our findings document that NHE-RF RNA expression varies greatly
among tissues (Fig. 6), being highest in liver, salivary gland, and
kidney, tissues with polarized epithelia rich in microvilli, and is
present in the mammary gland at a level about 60% that in the kidney.
The mammary gland thus contains a substantial amount of NHE-RF.
However, as the data derive from pooled tissues in which the
individuals’ estrogen status is unknown, these studies merely identify
the mammary gland as a tissue fairly rich in NHE-RF. Further studies
will be needed to document estrogen regulation of NHE-RF in normal
and/or malignant mammary gland as well as in other ER-containing
tissues in females and males.
|
|
Discussion |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Initial database searches using the product from the differential display revealed high sequence identity with the rabbit protein cofactor, known as NHE-RF, that is involved in the cAMP-dependent protein kinase A (PKA) inhibition of the Na+/H+ exchanger in the rabbit renal brush border membrane (5). More recently, the human homolog of NHE-RF that we identified was also described by another group and renamed EBP-50 by them, as it was shown by them to be an ezrin-radixin-moesin (ERM) binding phosphoprotein involved with the dynamics of cell ultrastructure (6). The ERM family of proteins plays structural and regulatory roles in assembly and stabilization of specialized plasma membrane domains, such as microvilli, membrane ruffles, and filopodia, by attaching the actin filaments of the cytoskeleton laterally to the plasma membrane. NHE-RF/EBP50, which was shown to directly interact with the ERM proteins (6), is believed to serve as an adapter protein between the ERM proteins and the integral membrane proteins. Interactions among NHE-RF/EBP50, the ERM proteins, and the Na+/H+ exchanger are believed to involve the two PDZ domains in NHE-RF/EBP50 that mediate protein-protein interactions and often assist in the formation of multiprotein complexes, particularly at the plasma membrane (17, 18, 19).
Our analyses in human tissues show that NHE-RF RNA is present at fairly high levels in the mammary gland and in other tissues containing a polarized epithelium and abundant microvilli, such as kidney, small intestine, liver, and salivary gland. As NHE-RF has clearly been shown to be localized in microvilli of cells expressing NHE-RF, it is intriguing that estrogens are known to induce cytoskeletal/ultrastructural changes in ER-containing breast cancer cells associated with increases in the number and length of microvilli at the cell surface (2, 20, 21, 22). These morphological changes in MCF-7 breast cancer cells were observable as early as 24 h after estradiol treatment, with the cells being uniformly covered with microvilli after 2 days of estrogen treatment (2). Similar effects were also demonstrated in other ER-positive breast cancer cells treated with estradiol, but not in an estrogen receptor-negative cell line (2). Related observations were also made in the anterior pituitary gland, where estrogens were found to markedly increase the number of microvilli (20). The findings of alterations in functioning of efferent ductules in the male reproductive tract of ER knockout (ERKO) mice (23) and the observation that the microvillus border of these estrogen target cells in ERKO mice is minimally developed and greatly disrupted (Hess, R., University of Illinois-Urbana, personal communication) support a role for ER in the development of microvilli and suggest that further investigation of the possible link between these abnormalities and a possible lack of ER-mediated production of NHE-RF is warranted. Future studies comparing NHE-RF function and regulation by estrogen in wild-type and ERKO mice should shed light on the role and importance of this protein in ER activities.
There is known to be extensive cross-talk among several pathways, including the cAMP/protein kinase A and epidermal growth factor pathways, that are involved in estrogen signaling (3, 4, 24), but the nature of the connections among these pathways is not well understood. NHE-RF might be one possible link through which the pathways communicate, as it has been shown to participate in several relevant aspects of signal transduction. NHE-RF exists in multiple phosphorylation states, has been shown to be regulated by protein kinase A in vitro, and is involved in protein kinase-mediated cell transduction pathways. Although the specific pathways associated with the E2-NHE-RF system have yet to be determined, the documentation of estrogen up-regulation of NHE-RF should assist in attempts to provide a greater understanding of the series of cellular events initiated by estrogen’s actions mediated by the ER.
|
Footnotes |
|---|
Received November 5, 1998.
|
References |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
-ergocryptine, estradiol, and thyrotropin-releasing hormone in
rat anterior pituitary cells in culture. J Cell Biol 86:377–387This article has been cited by other articles:
 |
|
 |
|
  L. Fouassier, P. Rosenberg, M. Mergey, B. Saubamea, A. Claperon, N. Kinnman, N. Chignard, G. Jacobsson-Ekman, B. Strandvik, C. Rey, et al. Ezrin-Radixin-Moesin-Binding Phosphoprotein (EBP50), an Estrogen-Inducible Scaffold Protein, Contributes to Biliary Epithelial Cell Proliferation Am. J. Pathol., March 1, 2009; 174(3): 869 - 880. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. D. Stender, J. Frasor, B. Komm, K. C. N. Chang, W. L. Kraus, and B. S. Katzenellenbogen Estrogen-Regulated Gene Networks in Human Breast Cancer Cells: Involvement of E2F1 in the Regulation of Cell Proliferation Mol. Endocrinol., September 1, 2007; 21(9): 2112 - 2123. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. A. Cardone, A. Bellizzi, G. Busco, E. J. Weinman, M. E. Dell'Aquila, V. Casavola, A. Azzariti, A. Mangia, A. Paradiso, and S. J. Reshkin The NHERF1 PDZ2 Domain Regulates PKA-RhoA-p38-mediated NHE1 Activation and Invasion in Breast Tumor Cells Mol. Biol. Cell, May 1, 2007; 18(5): 1768 - 1780. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Kujala, S. Hihnala, J. Tienari, K. Kaunisto, J. Hastbacka, C. Holmberg, J. Kere, and P. Hoglund Expression of ion transport-associated proteins in human efferent and epididymal ducts Reproduction, April 1, 2007; 133(4): 775 - 784. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. S. Carroll and M. Brown Estrogen Receptor Target Gene: An Evolving Concept Mol. Endocrinol., August 1, 2006; 20(8): 1707 - 1714. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Y. Kim, E. M. Woo, Y. T. E. Chong, D. R. Homenko, and W. L. Kraus Acetylation of Estrogen Receptor {alpha} by p300 at Lysines 266 and 268 Enhances the Deoxyribonucleic Acid Binding and Transactivation Activities of the Receptor Mol. Endocrinol., July 1, 2006; 20(7): 1479 - 1493. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Rai, A. Frolova, J. Frasor, A. E. Carpenter, and B. S. Katzenellenbogen Distinctive Actions of Membrane-Targeted Versus Nuclear Localized Estrogen Receptors in Breast Cancer Cells Mol. Endocrinol., June 1, 2005; 19(6): 1606 - 1617. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. I. Gorodeski, U. Hopfer, C. C. Liu, and E. Margles Estrogen Acidifies Vaginal pH by Up-Regulation of Proton Secretion via the Apical Membrane of Vaginal-Ectocervical Epithelial Cells Endocrinology, February 1, 2005; 146(2): 816 - 824. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Stossi, D. H. Barnett, J. Frasor, B. Komm, C. R. Lyttle, and B. S. Katzenellenbogen Transcriptional Profiling of Estrogen-Regulated Gene Expression via Estrogen Receptor (ER) {alpha} or ER{beta} in Human Osteosarcoma Cells: Distinct and Common Target Genes for These Receptors Endocrinology, July 1, 2004; 145(7): 3473 - 3486. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Incerpi, S. D'Arezzo, M. Marino, R. Musanti, V. Pallottini, A. Pascolini, and A. Trentalance Short-Term Activation by Low 17{beta}-Estradiol Concentrations of the Na+/H+ Exchanger in Rat Aortic Smooth Muscle Cells: Physiopathological Implications Endocrinology, October 1, 2003; 144(10): 4315 - 4324. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Pedram, M. Razandi, M. Aitkenhead, C. C. W. Hughes, and E. R. Levin Integration of the Non-genomic and Genomic Actions of Estrogen. MEMBRANE-INITIATED SIGNALING BY STEROID TO TRANSCRIPTION AND CELL BIOLOGY J. Biol. Chem., December 20, 2002; 277(52): 50768 - 50775. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Gartzke and K. Lange Cellular target of weak magnetic fields: ionic conduction along actin filaments of microvilli Am J Physiol Cell Physiol, November 1, 2002; 283(5): C1333 - C1346. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Shenolikar, J. W. Voltz, C. M. Minkoff, J. B. Wade, and E. J. Weinman Targeted disruption of the mouse NHERF-1 gene promotes internalization of proximal tubule sodium-phosphate cotransporter type IIa and renal phosphate wasting PNAS, August 20, 2002; 99(17): 11470 - 11475. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. R. Ediger, S.-E. Park, and B. S. Katzenellenbogen Estrogen Receptor Inducibility of the Human Na+/H+ Exchanger Regulatory Factor/Ezrin-Radixin-Moesin Binding Protein 50 (NHE-RF/EBP50) Gene Involving Multiple Half-Estrogen Response Elements Mol. Endocrinol., August 1, 2002; 16(8): 1828 - 1839. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. A. Oliveira, Q. Zhou, K. Carnes, R. Nie, D. E. Kuehl, G. L. Jackson, L. R. Franca, M. Nakai, and R. A. Hess ER Function in the Adult Male Rat: Short- and Long-Term Effects of the Antiestrogen ICI 182,780 on the Testis and Efferent Ductules, without Changes in Testosterone Endocrinology, June 1, 2002; 143(6): 2399 - 2409. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Karthikeyan, T. Leung, and J. A. A. Ladias Structural Determinants of the Na+/H+ Exchanger Regulatory Factor Interaction with the beta 2 Adrenergic and Platelet-derived Growth Factor Receptors J. Biol. Chem., May 17, 2002; 277(21): 18973 - 18978. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Sun, Y. R. Huang, W. R. Harrington, S. Sheng, J. A. Katzenellenbogen, and B. S. Katzenellenbogen Antagonists Selective for Estrogen Receptor {alpha} Endocrinology, March 1, 2002; 143(3): 941 - 947. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Lange Role of microvillar cell surfaces in the regulation of glucose uptake and organization of energy metabolism Am J Physiol Cell Physiol, January 1, 2002; 282(1): C1 - C26. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Q. Zhou, L. Clarke, R. Nie, K. Carnes, L.-W. Lai, Y.-H. H. Lien, A. Verkman, D. Lubahn, J. S. Fisher, B. S. Katzenellenbogen, et al. Estrogen action and male fertility: Roles of the sodium/hydrogen exchanger-3 and fluid reabsorption in reproductive tract function PNAS, October 31, 2001; (2001) 241245898. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. J. Pepe, M. G. Burch, and E. D. Albrecht Estrogen Regulates 11{beta}-Hydroxysteroid Dehydrogenase-1 and -2 Localization in Placental Syncytiotrophoblast in the Second Half of Primate Pregnancy Endocrinology, October 1, 2001; 142(10): 4496 - 4503. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. J. Pepe, M. G. Burch, C. P. Sibley, W. A. Davies, and E. D. Albrecht Expression of the mRNAs and Proteins for the Na+/H+ Exchangers and Their Regulatory Factors in Baboon and Human Placental Syncytiotrophoblast Endocrinology, August 1, 2001; 142(8): 3685 - 3692. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. O. Stemmer-Rachamimov, T. Wiederhold, G. P. Nielsen, M. James, D. Pinney-Michalowski, J. E. Roy, W. A. Cohen, V. Ramesh, and D. N. Louis NHE-RF, a Merlin-Interacting Protein, Is Primarily Expressed in Luminal Epithelia, Proliferative Endometrium, and Estrogen Receptor-Positive Breast Carcinomas Am. J. Pathol., January 1, 2001; 158(1): 57 - 62. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Maudsley, A. M. Zamah, N. Rahman, J. T. Blitzer, L. M. Luttrell, R. J. Lefkowitz, and R. A. Hall Platelet-Derived Growth Factor Receptor Association with Na+/H+ Exchanger Regulatory Factor Potentiates Receptor Activity Mol. Cell. Biol., November 15, 2000; 20(22): 8352 - 8363. [Abstract] [Full Text]
|
|
|
|
 |
|
 M. G. Ghosh, D. A. Thompson, and R. J. Weigel PDZK1 and GREB1 Are Estrogen-regulated Genes Expressed in Hormone-responsive Breast Cancer1, 2 Cancer Res., November 1, 2000; 60(22): 6367 - 6375. [Abstract] [Full Text]
|
|
|
|
 |
|
 C. H Mitchell, K. Peterson-Yantorno, M. Coca-Prados, and M. M Civan Tamoxifen and ATP synergistically activate Cl- release by cultured bovine pigmented ciliary epithelial cells J. Physiol., May 15, 2000; 525(1): 183 - 193. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. M. Bigsby, H. Cardenas, A. Caperell-Grant, and C. J. Grubbs Protective effects of estrogen in a rat model of age-related cataracts PNAS, August 3, 1999; 96(16): 9328 - 9332. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Q. Zhou, L. Clarke, R. Nie, K. Carnes, L.-W. Lai, Y.-H. H. Lien, A. Verkman, D. Lubahn, J. S. Fisher, B. S. Katzenellenbogen, et al. From the Cover: Estrogen action and male fertility: Roles of the sodium/hydrogen exchanger-3 and fluid reabsorption in reproductive tract function PNAS, November 20, 2001; 98(24): 14132 - 14137. [Abstract] [Full Text] [PDF]
|
|
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| Endocrinology | Endocrine Reviews | J. Clin. End. & Metab. |
| Molecular Endocrinology | Recent Prog. Horm. Res. | All Endocrine Journals |
