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antisense decreases brain estrogen receptor levels and
affects ventilation in male and female rats
Division of Basic Biomedical Sciences, University of South Dakota School of Medicine, Vermillion, South Dakota 57069
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ABSTRACT |
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 TOP
 ABSTRACT
 INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
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We hypothesized that administration of an antisense
oligodeoxynucleotide (ODN) to estrogen receptor (ER)-
mRNA decreases the ER protein in the neonatal rat brain, alters the sex-specific ventilatory responses to aspartic acid in rats, and counteracts the
effects of testosterone proportionate (TP) in females. One-day-old rat
pups were injected intraventricularly with vehicle, antisense ER ODN,
or scrambled ODN control. Additional groups of females received TP or
vehicle and one of the three treatments. Brain ER protein levels were
decreased by 65% at 6 h and 35% at 24 h after antisense
ODN. Aspartic acid decreased ventilation in all groups of weanling
males and females except ER ODN-treated females and TP-vehicle-treated
females. Aspartic acid decreased ventilation in all groups of adult
females except those given TP and in males. Weanling ER ODN-treated
rats were shorter and weighed less than controls. Only adult ER
ODN-treated males exhibited these traits. Thus neonatal ER affects
aspartic acid modulation of breathing and body growth in a sex-specific
and developmental manner.
cortex; hypothalamus; brain stem; N-methyl-D-aspartic receptor; aspartic acid; body weight; length
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INTRODUCTION |
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TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
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ESTROGEN RECEPTORS (ER) modulate the organization and activation of the developing rat brain (2). During the late fetal and early neonatal period, ER are more widely and abundantly expressed compared with during adulthood (7, 19, 29). This is of great importance, because these early times coincide with the critical developmental period when major organizational changes take place in the neuronal circuitry of the rat brain (19, 29). Thus changes in ER expression during this stage can influence the growth and pattern of differentiation of the developing rat brain and may have enduring consequences into adulthood. Methods to study the role of ER in the developing rat brain include the use of ER antagonists, aromatase inhibitors, gonadectomy, and genetic models such as transgenic knockouts (6, 16, 24, 25). A novel molecular technique involves the use of antisense oligodeoxynucleotide (ODN) against ER. Antisense ODNs are short sequences of DNA or RNA, usually 12-15 bases in length, that are complementary to a specific gene and inhibit the expression of the gene and thus the protein that would normally be produced.
Central administration of an antisense ODN against ER- mRNA into
3-day-old pups results in alterations of behavior and the volume of the
sexually dimorphic nuclei in the adult (20, 21). Whether
these changes were due to antisense ODN effects on ER protein
production in the neonatal rat brain was not demonstrated. Moreover, we
were interested in the role that ER has in modulating ventilation in
response to systemic administration of aspartic acid, an
N-methyl-D-aspartic (NMDA) receptor agonist.
Previously, our laboratory has shown that treatment of neonatal male
rats with estradiol benzoate reverses the male-specific ventilatory response to aspartic acid (28). In earlier studies, our
laboratory showed that adult males who had been castrated after puberty
or received testosterone propionate (TP) shortly after birth did not
lose their malelike ventilatory response to aspartic acid (see
references in Ref. 28). These results suggested that
hypogandodism itself was not sufficient to alter the malelike
ventilatory response to aspartic acid, but that "feminization" of
the brain by neonatal estradiol benzoate was necessary. In addition,
adult female rats treated neonatally with TP exhibited malelike
ventilatory responses to aspartic acid (27). We
hypothesized that these agents may be acting through ER because TP may
be concerted to ER in the brain. The present study was designed to test
the hypothesis that depression of ER protein production in the brain of
neonatal rats would alter the sex-specific ventilatory response to
aspartic acid and counteract the effects of TP. To determine whether
the activational effects of puberty modified these responses to
aspartic acid, we evaluated animals shortly after weaning (~23 days
of age) and at adulthood (2-3 mo old).
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METHODS |
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TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
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Animals and groups. Male and female Sprague-Dawley rats (proven breeders), obtained from Hilltop, Scottsdale, PA, were bred in our animal housing facility. Food and water were available ad libitum. Lights were maintained on a 12 h on-12 h off cycle. The Institutional Animal Care and Use Committee of the University of South Dakota approved all procedures.
One day after birth, rat pups were divided by sex and assigned to nine groups. These consisted of male and female rat pups who were treated centrally with 1) the vehicle, sesame oil; 2) a 15-mer scrambled ODN against the-isoform of
the ER mRNA; 3) a 15-mer antisense ODN against the
-isoform of the ER mRNA; and female rats treated first with one of
the three treatments mentioned above and then with 50 µg of TP
suspended in 0.1 ml of sesame oil.
Intraventricular infusions.
One-day-old male and female rat pups were cold anesthetized and placed
in a stereotaxic apparatus with a modified rat head holder. After an
incision was made in the scalp to expose the bregma, the rat pups were
injected intraventricularly with either the vehicle sesame oil, a
scrambled ODN against the ER mRNA, or an antisense ODN against the ER
mRNA. The volume of injection was 1 µl bilaterally, and the
concentration of the scrambled and the antisense ODN was 1 µg
DNA/µl sesame oil. To ensure a homogenous distribution of the ODN in
the sesame oil, the ODN were sonicated for 5 min before injection. The
sequence of the antisense ODN was 5' CAT GGT CAT GGT CAG 3', and it
spanned the putative translation start codon for the -isoform of the
rat ER mRNA. The scrambled ODN consisted of the same 15 bases as the
antisense ODN but in a scrambled order that has no homology to
sequences of mRNA submitted to various genomic databases including
GenBank. The sequence for the scrambled ODN was 5' ATC GTG GAT CGT GAC
3'. The unmodified ODN were obtained from Integrated DNA Technologies
(Coralville, IA).
Tissue collection and homogenization.
At 6 or 24 h after the central injection, the rat pups treated
with either vehicle, scrambled ODN, or antisense ODN were killed by
decapitation. For each of the 6-h and the 24-h samples, respectively, seven males and eight females were treated with vehicle, five males and
seven females were treated with the scrambled ODN, and seven males and
eight females were treated with the antisense ODN. To collect the
brains, an incision was made in the scalp and the whole brain was
extracted. The cortex, hypothalamus, and brain stem were dissected on
ice and then frozen at 
70°C. The samples were subsequently minced
and homogenized in a Dounce homogenizer at 1:5 (wt/vol) in a
homogenization buffer containing 20 mM Tris · HCl (pH 7.4), 2 mM EDTA, 5 mM EGTA, 0.25 M sucrose, 50 mM 2-mercaptoethanol, 1%
Nonidet-40, and 1 mM phenylmethylsulfonyl fluoride, a protease inhibitor. The homogenate was sonicated for 10 s and incubated for
90 min on ice to ensure extraction of all proteins into the detergent-soluble fraction. The samples were then centrifuged at 16,000 g for 15 min. The supernatant consisting of the cytosolic and nuclear extracts was collected, cryofrozen, and stored at 70°C.
Protein detection using Western blot analysis. Western blot analysis was used to examine the protein content of ER in the neonatal rat brain following the ODN treatments (10). The protein concentration of the samples was determined using the Pierce bicinchoninic acid protein assay (Rockford, IL). Cytosolic protein (30 µg) was loaded in each lane of a polyacrylamide gel and separated by electrophoresis. From preliminary experiments, this concentration was determined to be in the linear range of the Western blot.
One gel with samples from the 6-h treatment group was silver stained to confirm the presence of equal amounts of protein in all the lanes. Polyclonal antibodies ER HC-20 and ER MC-20 (1:100) (Santa Cruz Biotechnology, Santa Cruz, CA) that recognize ER- were used. The ER
protein bands were visualized with enhanced chemiluminescence
development (Amersham, Arlington Heights, IL) of the PVDF membrane
followed by exposure to X-ray film.
The ER--specific bands were quantified by densitometric analysis of
the exposed films by using a Molecular Dynamics densitometer (Sunnyvale, CA) and were expressed as arbitrary densitometric units.
Differences between the ER bands of either the antisense ODN-treated or
the scrambled ODN-treated samples and the vehicle-treated samples were calculated.
Ventilatory measurements. Ventilation was evaluated by using the barometric plethysmographic techniques as previously described by Schlenker and colleagues (27). Awake, unrestrained rats were placed into a cylindrical Plexiglas chamber (20 cm long and 8 cm in diameter for the weanlings and 25 cm long and 14 cm in diameter for the adult rats). Each rat was weighed and afterward received a 0.1- to 0.3-ml injection of saline depending on age. The rat was then placed into the chamber, and ventilation was evaluated 30 min later. Subsequently, the rat was removed and injected with 580 mg/kg aspartic acid (Sigma Laboratories, St. Louis, MO) with the same volume as the saline injection. The rat was again placed into the chamber, and ventilation was evaluated after 30 min. Ventilatory parameters that were evaluated included the tidal volume, frequency of breathing, and minute ventilation.
Measurements of body length. In addition to weighing rats, we also evaluated body lengths (nose-to-anus length). These measurements were made to determine whether ER perturbations could affect growth (length), a sex-specific parameter that is affected by neonatal hormonal perturbations.
Statistical analysis. To determine whether there was a significant effect of ER ODN on ER protein levels, a Wilcoxon's signed-rank test was used. The effects of ER ODN relative to control groups (scrambled ODN and vehicle) on body weight and body length were computed by using a one-way analysis of variance by sex at each age (weanling and adult). To determine whether aspartic acid affected ventilation relative to saline, a Student's t-test was used for each treatment group. The intent of this test was just to determine whether an effect was present, not to compare the magnitude of responses among the various groups. Significance was accepted at P < 0.05.
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RESULTS |
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TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
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Effects of ER ODN on
ER protein levels.
ER were present in the cortex, hypothalamus, and brain stem of the
neonatal rat pups (Figs. 1 and
2). No significant sex differences were noted in the ER protein levels in any of the three brain regions
from the control or the treated rats (data not shown). Therefore,
results obtained from the male and female neonatal rats were pooled.
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protein
content in the three brain regions was significantly decreased
(P < 0.02) relative to vehicle-treated neonatal rats (Fig. 1). Complete blockade of ER- protein expression in the three
brain regions with antisense ODN treatment was noted in 47% of the gels.
Twenty-four hours after central administration of antisense ODN, the ER
protein content in cortex, hypothalamus, and brain stem was
significantly decreased relative to the vehicle-treated neonatal rats
(P < 0.05, Fig. 2). However, the decrease in ER protein levels with antisense ODN-treatment relative to vehicle treatment was greater at 6 h (65%) than at 24 h after
treatment (35%). Relative to vehicle, administration of scrambled ODN
did not significantly affect the ER protein content in any of the three
brain regions at either 6 or 24 h after treatment (Figs. 1B and 2B).
Effects of ER ODN on body weights and
lengths.
In Fig. 3, the effect of ER ODN on body
weights of weanling and adult rats is presented. Both weanling male and
female ER ODN-treated rats have significantly (P < 0.01) lower body weights than control sex-matched animals. There was no
significant difference in body weight between the scrambled ODN and
vehicle controls. Weanling rats treated with TP and ER-ODN weighed
significantly (P < 0.01) more than rats in the two
other TP-treated groups. In adulthood, ER ODN-treated males were still
lighter than controls, but there was no longer a difference among
females treated with vehicle, antisense ODN, scrambled ODN, or TP and
antisense ODN. In contrast, body weights were increased in TP-treated
vehicle and scrambled ODN rats. The effects of the neonatal treatments on body length were identical to those reported for body weight for
each sex (data not shown). The only exception was that body length was
longer in vehicle and TP-treated adult females relative to the
other female treatment groups.
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Effects of ER ODN on aspartic acid
modulation of breathing.
The effects of aspartic acid on ventilation are presented in Figs.
4-6.
In weanling males, all treatments with aspartic acid significantly
(P < 0.05) depressed ventilation relative to saline treatment. In contrast, weanling ER ODN females exhibited no response to aspartic acid relative to saline. Whereas the two control groups of
TP-treated females exhibited no ventilatory response to aspartic acid,
the TP- and antisense ODN-treated females exhibited a depression of
breathing in response to aspartic acid relative to saline responses.
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DISCUSSION |
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TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
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This study demonstrated that ER- was found abundantly in the
cortex, hypothalamus, and brain stem of the neonatal rat. The antisense
ODN against the ER mRNA, but not the control treatments, decreased the
ER protein levels at both 6 and 24 h after administration, with
the inhibition of ER- protein production being greater at 6 than at
24 h. There were no sex differences in the levels of ER protein
for either the control or the ODN-treated rats in any of the three
brain areas studied. Physiologically, ER ODN treatment decreased body
weights and lengths and affected ventilatory responses to aspartic acid
in weanlings in a sex-specific manner. Treatment of female rats with TP
plus ER ODN reversed the effects of ER ODN alone of weanlings in
response to aspartic acid and the decrease in body length and weight.
However, some of these effects were reversed in adulthood.
Effects of ER ODN on
ER protein production.
This is the first study to demonstrate that administration of antisense
ODN against ER- mRNA to the neonatal rat brain can block ER-
protein production in the cortex, hypothalamus, and brain stem, as
measured by the Western blot technique. Total blockade of ER-
protein production was seen in 47% of all samples at 6 h after
administration of the antisense ODN. ER- protein content in the
three brain regions was also significantly decreased 24 h after
administration of antisense ODN relative to the vehicle control, but
not to the extent it had been at 6 h.
protein content at 6 h
relative to 24 h after the central administration of antisense ODN
helps denote the time course of action of the antisense ODN. The data
suggest an onset of activity 4-6 h after administration. This
agrees with the known mode of action of the antisense ODN to block
transcription and protein production (1, 17, 32). The
apparent decreased effectiveness of the antisense ODN 24 h after
its administration may be due to antisense ODN degradation, which then
allows renewed synthesis of ER-. These data also confirm the
reversibility of the antisense technique and the lack of permanent damage or toxicity using unmodified antisense ODN. Moreover, these results suggest that dynamic turnover of ER protein production in the
brain occurs during this sensitive developmental period.
As demonstrated by this and other studies (3, 21, 24, 35),
the effects of a single dose of antisense ODN within the brain can be
effective and reversible. The lack of toxicity observed with the use of
the unmodified antisense ODN against ER- further confirms the
usefulness of this technique for in vivo studies. Furthermore, the
brain itself may constitute a unique environment in which unmodified
antisense ODN can exert their effects because of the relative stability
of these molecules in the central nervous system, or, possibly, because
of an extremely efficient uptake into the neuronal cells
(3). Thus the distribution and uptake of the ODN
in the brain may be significantly better than that predicted from cell
culture studies. Sommer and colleagues (30) used
radiolabeled or FITC-labeled ODN to demonstrated that these oligos
rapidly gain access into the neurons and remain intact for as long as
24 h postinjection. Moreover, after central injection of the ODN,
uptake has been observed at sites distant from the point of injection
(20, 30). Our results in the cortex, a region distant from
the ventricles, support this finding. Another important consideration
with the use of ODN is the possibility of toxic effects after exposure
to large concentrations of DNA. Unmodified ODN, as used in this study,
exhibit low levels of toxicity or hyperirritability (20, 32,
35). In contrast, ODN with modifications such as the addition of
sulfur or an amide group are associated with a high level of
dose-dependent toxicity and mortality (17).
Another result of this study was a lack of sex differences in the ER
protein levels in any brain region. However, the present study focused
on the ER protein levels in relatively large brain regions. We did not
look at specific brain nuclei that have been reported to have a
sexually dimorphic distribution of ER. For example, Kuhnemann and
colleagues (13) measured the ER in the various brain
nuclei in neonatal rats by quantitative in vitro autoradiography. Sex
differences in ER were noted in the medial preoptic nucleus and the
paraventricular nucleus as early as 24 h after birth. In contrast,
sex differences in ER in the ventromedial nucleus appeared much later,
between 5 and 10 days of age. Thus the asynchronous development of sex
differences in ER levels in specific brain nuclei suggests the
possibility of different "critical periods" for ER expression, and
therefore ER effects exist in various brain regions (8, 16, 18,
21).
The present study focused on the effects of blocking ER- protein
production. Because ER- may form dimers with isoforms of the
recently cloned ER-
subtypes, disruption of the activity of
dimers may have occurred (13, 15, 26). Furthermore, these two isoforms of the ER have been localized in different brain nuclei in
the rat and act through separate mechanisms, and thus they could have
different molecular and physiological effects (14, 23,
29).
Physiological consequences of ER ODN. Physiologically, we found effects of ER ODN on body weight, length, and aspartic acid modulation of ventilation. Of interest is that the findings in weanlings did not always persist in adults. This is particularly true for the female rats and indicates an activational effect of puberty that is sex specific. In a previous study in which ER ODN was administered to 3-day-old rat pups, there was no effect on body weight, suggesting that there is a critical time period for ER to affect neural systems associated with body weight maintenance (21). These systems may modulate feeding behavior, body temperature, fat deposition, and/or metabolism (5, 22). Neuromodulators regulated by ER that affect growth and development include galanin, insulin-like growth factor I (IGF-I), growth hormone-releasing hormone, thyroid hormone, and somatostatin (4, 11, 15). In the present study, we did not observe a difference in body temperature or metabolism in weanling or adult rats of either sex (data not presented). Future studies are needed to investigate the effect of ER ODN treatment on food intake and the central factors that underlie ER modulate of neurotransmitters associated with growth and development in weanlings and adults rats.
Ventilation in response to aspartic acid was markedly affected in female weanling rats treated with ER ODN. This suggests that there was an interaction between ER and NMDA receptors at a critical period in development that is sex specific (4). Watanabe and co-workers (33) noted that a specific subunit of the NMDA receptor, the NR2D, contains four estrogen-responsive elements. Moreover, within the hypothalamus, NR2D mRNA is colocalized with ER (33). This subunit is also prominently expressed within pontine and medullary nuclei associated with control of ventilation and is modulated in a developmental manner (9, 12, 34). Sex differences in the development of the NDR2 subunit need to be evaluated further to determine whether this may play a role in the sex-specific effects of aspartic acid noted in the present study.Effects of TP. Another aim of this study was to determine whether ER ODN pretreatment could alter the effects of TP administration. Neonatal androgenization with TP causes a transient increase in estrogen production centrally (due to aromatization of testosterone to estrogen, Ref. 31) and a longer term downregulation of estrogen receptors (18). By administration of ER ODN, which decreased ER during a 24-h window, the effects of excess estrogen due to TP may not have affected neurons and glial cells containing ER. Of interest, however, is that during adulthood this protective effect was no longer present. In support of this supposition we noted that adult females treated with neonatally with TP independent of central (ER ODN, scrambled ODN, or vehicle) neonatal treatments exhibited ventilatory responses to aspartic acid similar to that of normal males. In contrast, weanling females pretreated with either vehicle or scrambled ODN plus TP exhibited adultlike male responses to aspartic acid, but females treated with TP and ER ODN exhibited a ventilatory response to aspartic acid similar to that of the weanling pattern of response. Thus, in weanlings, ER ODN administered neonatally counteracted the effects of TP on aspartic acid modulation of breathing. A similar effect of TP and ER ODN was noted in weanlings for body weight and length. These results suggest that neuronal function associated before puberty is very different than after puberty. Preliminary data do indicate that neonatal treatment of female rats with TP results in higher leptin levels than treatment with vehicle (3.11 ± 0.26 ng/dl with TP compared with 2.19 ± 0.24 ng/dl vehicle, P < 0.02; unpublished observations). What the role of altering ER brain levels has on leptin levels or other neuromodulators such as NMDA receptors in male and female weanling and adult rats needs to be investigated.
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ACKNOWLEDGEMENTS |
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We thank Dr. Leo Kretzner for invaluable help in the development of this project.
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FOOTNOTES |
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These studies were supported in part by National Institute of Child Health & Human Development Grant HD-30393 (to E. H. Schlenker) and by NSF-EPSCor Grant OSR-9452894 (to K. M. Eyster).
Address for reprint requests and other correspondence: E. H. Schlenker, Division of Basic Biomedical Sciences, Univ. of South Dakota School of Medicine, 414 E. Clark St., Vermillion, SD 57069 (E-mail: eschlenk{at}usd.edu).
The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
Received 17 April 2001; accepted in final form 18 June 2001.
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REFERENCES |
|---|
|
TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
|
|---|
1.
Akhtar, S,
and
Agrawal S.
In vivo studies with antisense oligonucleotides.
Trends Pharmacol Sci
18:
12-18,
1997[Medline].
2.
Arnold, AP,
and
Gorski RA.
Gonadal steroid induction of structural sex differences in the central nervous system.
Annu Rev Neurosci
7:
413-442,
1984[ISI][Medline].
3.
Auger, AP,
Tetel MJ,
and
McCarthy MM.
Steroid receptor coactivator-1 (SRC-1) mediates the development of sex-specific brain morphology and behavior.
Proc Natl Acad Sci USA
97:
7551-7555,
2000
4.
Carbone, S,
Szwarcfarb B,
Losada M,
and
Moguilevasky JA.
Effect of ovarian hormones on the hypothalamic excitatory amino acids during sexual maturation in female rats.
Neuroendocrinology
62:
235-242,
1995.
5.
Czaja, CA.
Sex differences in the activational effects of gonadal hormones on food intake and body weight.
Physiol Behav
33:
553-558,
1984[Medline].
6.
Clancy, AN,
Zumpe D,
and
Michael RP.
Intracerebral infusion of an aromatase inhibitor, sexual behavior and brain estrogen receptor-like immunoreactivity in intact male rats.
Neuroendocrinology
61:
98-111,
1995[ISI][Medline].
7.
DonCarlos, LL,
and
Handa RJ.
Developmental profile of estrogen receptor mRNA in the preoptic area of male and female neonatal rats.
Dev Brain Res
79:
283-289,
1994[Medline].
8.
DonCarlos, LL,
McAbee M,
Ramer-Quinn DS,
and
Stancik DM.
Estrogen receptor mRNA levels in the preoptic area of neonatal rats are responsive to hormonal manipulation.
Dev Brain Res
84:
253-260,
1995[Medline].
9.
Dunah, AW,
Luo J,
Wang Y-H,
Yaduda RP,
and
Wolfe BB.
Subunit composition of N-methyl-D-aspartate receptors in the central nervous system that contain the NR2D subunit.
Mol Pharmacol
53:
429-437,
1998
10.
Eyster, KM,
Berger TL,
Rodrigo MC,
and
Sheth MV.
Protein phosphatase activity in the rat ovary throughout pregnancy and pseudopregnancy.
Biol Reprod
58:
338-345,
1998
11.
Garcia-Segura, LM,
Azcoitia I,
and
DonCarlos LL.
Neuroprotection by estradiol.
Prog Neurobiol
63:
29-60,
2001[ISI][Medline].
12.
Guthmann, A,
and
Herbert H.
Expression of N-methyl-D-aspartate receptor subunits in the rat parabrachial and Kölliker-Fuse nuclei and the selected pontomedullary brainstem nuclei.
J Comp Neurol
415:
501-517,
1999[ISI][Medline].
13.
Kuhnemann, S,
Brown TJ,
Hochberg RB,
and
MacLusky NJ.
Sex differences in the development of estrogen receptors in the rat brain.
Horm Behav
28:
483-491,
1994[Medline].
14.
Kuiper, GG,
Carlsson B,
Grandien K,
Enmark E,
Haggblad J,
Nilsson S,
and
Gustafsson JA.
Comparison of the ligand binding specificity and transcript tissue distribution of estrogen receptors and .
Endocrinology
138:
863-870,
1997
15.
Lebowitiz, SF,
Akabayashi A,
Alexander JT,
and
Wang J.
Gonadal steroids and hypothalamic galanin and neuropeptide Y: role in eating behavior and body weight control in female rats.
Endocrinology
139:
1771-1780,
1998
16.
Lewis, C,
McEwen BS,
and
Frankfurt M.
Estrogen-induction of dendritic spines in ventromedial hypothalamus and hippocampus: effects of neonatal aromatase blockade and adult GDX.
Dev Brain Res
87:
91-95,
1995[Medline].
17.
Liebert, MA.
Problems in interpretation of data derived from in vitro and in vivo use of antisense oligodeoxynucleotides.
Antisense Res Dev
4:
67-69,
1994[ISI][Medline].
18.
Mclusky, NJ,
Bowlby DA,
Brown TJ,
Peterson RE,
and
Hochberg RB.
Sex and the developing brain: suppression of neuronal estrogen sensitivity by developmental androgen exposure.
Neurochem Res
11:
1395-1414,
1997.
19.
MacLusky, NJ,
Chaptal C,
and
McEwen BS.
The development of estrogen receptor systems in the rat brain and pituitary: postnatal development.
Brain Res
178:
143-160,
1979[ISI][Medline].
20.
McCarthy, MM,
Brooks PJ,
Pfaus JG,
Brown HE,
Flanagan LM,
Schwartz-Giblin S,
and
Pfaff DW.
Antisense oligodeoxynucleotides in behavioral neuroscience.
Neuroprotocols
2:
1-8,
1993.
21.
McCarthy, MM,
Schlenker EH,
and
Pfaff DW.
Enduring consequences of neonatal treatment with antisense oligodeoxynucleotides to estrogen receptor mRNA on sexual differentiation of rat brain.
Endocrinology
133:
433-439,
1995[Abstract].
22.
Muramatsu, M,
and
Inoue S.
Estrogen receptors: how do they control reproductive and nonreproductive functions?
Biochem Biophys Res Commun
270:
1-10,
2000[ISI][Medline].
23.
Nilsen, J,
Mor G,
and
Naftolin F.
Estrogen-regulated developmental neuronal apoptosis is determined by estrogen receptor subtype and the Fas/Fas ligand system.
J Neurobiol
43:
64-78,
2000[ISI][Medline].
24.
Ogawa, S,
Olazabal UE,
Parhar IS,
and
Pfaff DW.
Effects of intrahypothalamic administration of antisense DNA for progesterone receptor mRNA on reproductive behavior and progesterone receptor immunoreactivity in female rat.
J Neurosci
14:
1766-1774,
1994[Abstract].
25.
Ogawa s Chan, J,
Chester AE,
Gustafsson JA,
Korach KS,
and
Pfaff DW.
Survival of reproductive behaviors in estrogen receptor gene-deficient (ERKO) male and female mice.
Proc Natl Acad Sci USA
96:
12887-12892,
1999
26.
Petersen, DN,
Tkalcevuc GT,
Koza-Taylor PH,
Turi TG,
and
Brown TA.
Identification of estrogen receptor 2, a functional variant of estrogen receptor expressed in normal rat tissues.
Endocrinology
139:
1082-1092,
1998
27.
Schlenker, EH,
Goldman M,
and
Holman G.
The effects of aspartic acid on ventilation in androgenized and ovariectomized female rats.
J Appl Physiol
72:
2255-2258,
1992
28.
Schlenker, EH,
Goldman M,
and
Walsh S.
The effect of perinatal estradiol benzoate, administration on control of ventilation in adult male rats.
Physiol Behav
52:
1113-1116,
1992[Medline].
29.
Sibug, RM,
Stumpf WE,
Shughrue PJ,
Hochberg RB,
and
Drews U.
Distribution of estrogen target sites in the 2-day-old mouse forebrain and pituitary gland during the `critical period' of sexual differentiation.
Dev Brain Res
61:
11-22,
1991.
30.
Sommer, W,
Xia C,
Erdmann B,
and
Fuxe K.
The spread and uptake pattern of intracerebrally administered oligonucleotides in nerve and glial cell populations of the rat brain.
Soc Neurosci Abstr
23:
1163,
1997.
31.
Stewart J, and Rajabi H. Estradiol derived from testosterone in
prenatal life effects the development of catecholamine systems in the
frontal cortex in the male rat. Brain Res 157-160,
1994.
32.
Stein, C,
and
Cohen JS.
Oligodeoxynucleotides as inhibitors of gene expression: a review.
Cancer Res
48:
2659-2668,
1988
33.
Watanabe, T,
Inoue S,
Hiroi H,
Orimo A,
and
Muramatsu M.
NMDA receptor type 2D gene as a target for estrogen receptor in the brain.
Mol Brain Res
63:
375-379,
1999[Medline].
34.
Wenzel, A,
Villa A,
Mohler H,
and
Benke D.
Developmental and regional expression of NMDA subtypes containing the NR2D subunit in rat brain.
J Neurochem
66:
1240-1248,
1996[ISI][Medline].
35.
Wielbo, D,
Sernia C,
Gyurko R,
and
Phillips MI.
Antisense inhibition of hypertension in the spontaneously hypertensive rat.
Hypertension
25:
314-319,
1995
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