 |
||||
|
||||
|
||||
|
||||
Previous Article | Next Article
Volume 16, Number 16, Issue of August 15, 1996 pp. 4823-4834
Copyright ©1996 Society for NeuroscienceDopaminergic Regulation of Progesterone Receptors: Brain D5 Dopamine Receptors Mediate Induction of Lordosis by D1-Like Agonists in Rats
Ede Marie Apostolakis1, Janos Garai2, Charles Fox3, Carolyn L. Smith1, Stanley J. Watson4, James H. Clark1, and Bert W. O'Malley1 1 Department of Cell Biology, Baylor College of Medicine, Houston, Texas 77030, 2 Department of Pathophysiology, University Medical School of Pécs, Pécs, Hungary, 3 Engineering Animation Inc., Ames, Iowa 50010, and 4 Mental Health Research Institute, University of Michigan, Ann Arbor, Michigan 48109
ABSTRACT
To characterize the signaling pathway by which the neurotransmitter dopamine modulates progesterone receptor (PR) activation, the steroid-dependent behavior lordosis was used in estrogen-primed ovariectomized Sprague-Dawley rats with stereotaxic implanted third ventricle cannulas. Lordosis was observed in response to solicitous males in females after central administration of the D1-like agonist SKF38393 and three of its analogs (SKF77434, SKF75640, and SKF85174). In contrast, D1-like antagonist SCH23390 and D1-like/D2 repopulation inhibitor EEDQ blocked behavior inducible by the D1-like agonists. Further, antisense oligonucleotides to D5, but not D1, dopamine receptor mRNA suppressed reproductive behavior associated with D1-like stimulation. This finding provides strong evidence that dopaminergic modulation of lordosis is mediated by the novel D5 dopamine receptor. Although D1, but not D5, dopamine receptor mRNAs were detected in the ventromedial nucleus (VMN) by in situ hybridization, agonists microinjected into the VMN, but not into the arcuate nucleus or preoptic area, induced lordosis, suggesting the functional presence of D5 dopamine receptors in the VMN. Also in support, D5 receptor mRNA antisense microinjected into the VMN blocked the subsequent induction of lordosis by D1-like agonists. Finally, facilitation of sex behavior by D1-like agonists was blocked by the antiprogestin RU38486 and PR antisense oligonucleotide. Collectively, the data provide strong evidence for dopaminergic modulation of reproductive behavior through D5 dopamine receptor-mediated modulation of PR-dependent behavior in rat CNS.
Key words: steroid receptors; D5 dopamine receptors; lordosis; antisense oligonucleotides; progesterone; estrogen
INTRODUCTION
In the female rat, the ovarian steroid hormone estrogen modulates the reproductive behavior lordosis (Pfaff and Schwartz-Giblin, 1988). Estrogen effect on lordosis is mediated in part by steroid receptors in the estrogen-concentrating neurons located in the ventrolateral region of the hypothalamic ventromedial nucleus (VMN). The major effect of estrogen on lordosis is thought to be mediated by progesterone receptors (PR), because (1) estrogen induces PR in the VMN, and (2) RU38486 and PR antisense oligonucleotide block the induction of lordosis by progesterone (P) (Pfaff and Schwartz-Giblin, 1988
As ligand-dependent nuclear transcription factors, steroid receptors alter the expression of specific genes or gene networks (Evans, 1988
Preliminary results of this study have been published in abstract form at the Annual Meeting of the Endocrinology Society (Anaheim, CA).
MATERIALS AND METHODS
Synthesis of designed oligodeoxynucleotides and materials. Sense (5to 3
For the rat D1 dopamine receptor (Zhou et al., 1990
For the rat D5 dopamine receptor (Tiberi et al., 1991
For the rat PR, the sense sequence was 5
Several selective dopaminergic agonists were used in the present studies. The D1-like agonists 1-phenyl-2,3,4,5-tetrahydro-benzazepine (SKF 38393) and N-allyl-benzazepine hydrochloride (SKF 77434), and the D1-like antagonist SCH23390 (Research Biochemicals, Natick, MA) were dissolved in sterile water. The D1 agonists 3-allyl-6-chloro-7,8-dihydroxy-1-2,3,4,5,-tetrahydro-11-1-benzazepine (SKF85174), SKF82958, and SKF75670 were kindly provided by SmithKline & Beecham (King of Prussia, PA) and also dissolved in water. The steroids 17
-estrogen benzoate (EB) and P (Sigma, St. Louis, MO), the antiprogestin RU38486 (Roussal-UCLAF, Paris, France), and the irreversible downregulator N-ethoxycarbonyl-2 ethoxy-1.2-dihydroquinoline (EEDQ) of D1-like and D2 repopulation were dissolved or suspended in sesame oil. For agonist control treatment in those animals cannulated in the arcuate nuclei (AN) or preoptic area (POA), gonadotropin-releasing hormone was kindly provided by W. W. Vale (The Salk Institute, La Jolla, CA) and suspended in sesame oil. All dopaminergic drugs were mixed not more than 1 hr before intracerebroventricular administration and given between 1 and 2 hr after lights on. Whenever possible, doses were based on published studies showing effective concentrations for desired effect; effective dose was then confirmed in the present study. Dose dependency was verified when appropriate.
Preparation of animals. Ovariectomized female rats (160-180 gm) were obtained from Sasco (Houston, TX), housed three per cage, and maintained on a 12:12 hr light/dark cycle (lights on at 0700 CST) with rat chow and water in excess ad libitum. After acclimation (5-7 d), animals were primed with EB (100 µg, s.c., at
44 hr), then treated with P (100 ng, s.c., at 0 hr), and screened for 10 min at 2-4 hr for receptive sex behavior to a series of two sexually active males housed individually in 50 × 45 × 24 cm polystyrene arenas. Females displaying lordosis underwent stereotaxic implantation of third ventricle cannula guides (26 gauge) (Plastics One, Roanoke, VA) as described previously (Mani et al., 1994
Behavioral testing lordosis in female rats. For each experiment, females were randomly assigned identification numbers after experimental treatment and at 1-4 hr later randomly tested during the light cycle for lordosis (LQ) with a series of four males for 5 min each. Prospective scoring was done by individuals blind to individual animal treatment. Tests for intertester and intratester reliability were conducted on a regular basis. Briefly, LQ was calculated as percent lordosis (immobile dorsiflexion of the female vertebral column with head at 45° angle from floor and perineal elevation preceded by flank contact with the male) per total mounts × 100. To provide maximal quantification and description, behavioral scores were assigned (0 = no dorsiflexion; 1 = slight dorsiflexion, some head elevation, back parallel to floor; 2 = moderate dorsiflexion with some leg extension from crouch position; 3 = full dorsiflexion). Only scores of 2 and 3 were judged positive for lordosis. Also noted were such receptive behaviors as acceptance and rejection. Finally, proceptive behaviors such as hopping, darting, ear wiggling, and approaching were recorded. To allow for females to act as their own controls, testing was conducted immediately before experimental treatment. All rats were tested before EB priming and excluded if they displayed false-positive responses (20% LQ).
Location studies. For those rats participating in the location studies, double cannulae were implanted in the VMN (26 gauge with 1.5 mm center to center of cannulae) and AN (26 gauge, 1.0 mm between cannulae) and POA (26 gauge, 1.5 mm between cannulae) according to the stereotaxic atlas of Paxinos and Watson (1988)
In situ hybridization study and probe preparation. Tissue preparation and in situ hybridization were performed as described previously (Fox et al., 1993
Studies for specificity of D1 dopamine receptor oligonucleotide specificity. Dopaminergic receptors in striatum and nucleus accumbens mediate levels of basal motor activity. Importantly, phenotypic behavior in targeted D1 mutant mice relative to their wild-type littermates include basal hyperactivity and failure of SKF82958 challenge to maximally enhance motor hyperactivity (Xu et al., 1994
Studies for D5 dopamine receptor oligonucleotide specificity. Using another clearly defined experimental system, the specificity of D5 dopamine receptor oligonucleotides was assessed in transient transfection experiments in mouse L(tk
5 cells per well were plated in a six-well culture dish in phenol red-free DMEM containing 5% charcoal-stripped FBS. Cells were then transferred to serum-free DMEM and infected with a variable amount of antisense or random sense D5 oligonucleotides (0-0.83 nmol/well) in the presence of 12.5 ng/well rat estrogen receptor (ER) expression vector (pJ3ratER) and 125 ng/well of a synthetic ER-responsive target gene (ERE-e1b-CAT). The ERE-e1b-CAT gene consists of the vitellogenin A2 estrogen response element linked upstream of the e1b TATA box and chloramphenicol acetyl transferase (CAT) reporter gene. A modified adenoviral system described by Cristiano et al. (1993)
Statistics. When animals served as their own controls, the significance of change in reproductive or motor behavior was assessed by two-way ANOVA with repeated measures (p
0.05 significant). Duncan's multiple range test was used for individual comparisons. When groups of animals were compared, one-way ANOVA, followed by the Mann-Whitney U test and Student's t test, was used. In the lordosis studies, the occurrence of proceptive behavior was altered in parallel with the lordotic activity and have not been statistically reported here. In the locomotor studies, the occurrence of sniffing, rearing, and grooming was consistent throughout all sessions regardless of treatment.
RESULTS
General behavior responses to EB
Animals receiving EB (subcutaneously) and P (subcutaneously or introcerebroventricularly) consistently exhibited high levels of proceptive (ear wiggling, hopping, and darting) and receptive (lordosis) behaviors in response to male mounting during the light cycle. Because excessively high doses of estrogen can induce lordosis in the absence of P (Pfaff and Schwartz-Giblin, 1988The high EB-priming dose used in this study afforded the opportunity for decreasing intracerebroventricular P dose toward more physiological concentrations. For positive controls and screening for false-negative animals, an intracerebroventricular P dose of 50 ng was chosen, because this dose was associated with the exhibition of proceptive and receptive behaviors comparable with those receiving lower EB (10 µg, s.c.) and higher P (2 mg, s.c.) doses. For all experiments, the data from animals treated with EB only that exhibited an LQ of 20% or above before experimental treatment (8 ± 2% of all animals) were excluded from analysis. Administration of vehicle alone failed to facilitate lordosis.
Dose response to D1-like dopamine agonists
It has already been demonstrated that the D1-like agonist SKF38393, but not its vehicle, induces lordosis in rats primed with EB (5-10 µg) (Foreman and Moss, 1979
Fig. 1. Ability of selective D1-like dopaminergic agonists to mediate reproductive behavior in ovariectomized rats with indwelling third ventricle cannulae. Microinjections of four selective D1-like agonists facilitated sex behavior in highly receptive rats primed with EB. Each experiment was repeated at least twice. The results are expressed as LQ (defined as percent of positive lordosis responses divided by number of mounts by a series of 4 male rats). Bars represent mean LQ ± SEM. Asterisks indicate a significant increase in LQ compared with the pretreatment response of the same rats 1 hr before D1-like agonist treatment (ANOVA, p
[View Larger Version of this Image (20K GIF file)]
Inhibition of behavioral responses by D1-like antagonists
Selective D1-like blockade and inhibition of dopamine receptor repopulation suppressed the induction of lordosis by D1-like agonist stimulation (Fig. 2). When animals were pretreated for 1 hr with the D1-like antagonist SCH23390, lordosis responses after microinjections of SKF38393 were blocked in a dose-dependent manner (Fig. 2, lane 7) (most effective dose). Likewise, animals pretreated for 24 hr with the irreversible D1-like/D2 antagonist EEDQ consistently failed to display receptive behavior after the dopaminergic agonist was administered (Fig. 2, lane 10). Hopping, darting, and ear wiggling also were inhibited by both antagonists, whereas rejection behaviors were common. In the presence or absence of EB, neither SCH23390 nor EEDQ facilitated the appearance of lordosis (Fig. 2, lanes 6, 9 for EB, and lanes 3, 8 for no EB pretreatment). Likewise, neither vehicle nor EB induced response (Fig. 2, lanes 1, 2).
Fig. 2. Inhibition of SKF-induced lordosis behavior by selective D1-like antagonists. Chronically cannulated rats primed with EB were given microinjections of the D1-like antagonist SCH23390 1 hr before or the irreversible D1-like/D2 antagonist EEDQ 24 hr before challenge with SKF38393. The animals were screened for nonspecific effect of antagonist before agonist challenge and analyzed as described in Figure 1 and in Materials and Methods.
[View Larger Version of this Image (9K GIF file)]
D1 and D5 dopamine receptor oligodeoxynucleotides, behavior, and transcription
Antisense oligonucleotide to rat D5, but not D1 dopamine receptor mRNA, effectively suppressed SKF induction of receptive (LQ) behavior (Fig. 3). Oligos were administered intracerebroventricularly 24 hr before agonist treatment. Commonly, females pretreated with antisense oligos to D5 (2 nM), but not D1 (2-4 nM), dopamine receptor mRNA aggressively rejected male solicitation (Fig. 3, lanes 11, 15, 19 for D5 AS, lane 7 for D1 AS). In contrast, females administered either random sense to D5 receptor mRNA (Fig. 3, lanes 9, 13, 17) or random sense or antisense to D1 receptor mRNA (Fig. 3, lanes 5, 7) exhibited high levels of receptive and proceptive behaviors when given D1-like agonists. No significant effect on reproductive behavior was observed for random sense and antisense oligos in the presence of only EB (Fig. 3, lanes 4, 6, 8, 10, 12, 14, 16, 18). For oligos to D1 dopamine receptor mRNA, no untoward effect was noted in animals. In contrast, a high dose (4 nM) of D5 antisense was associated with the death of several animals within 48 hr of oligo treatment. A lower dose (1 nM) of D5 antisense was characterized by irregular displays of lordosis and frequent, aggressive rejection behaviors when challenged with D1-like agonists.
Fig. 3. Inhibition of SKF-facilitated sex behavior by phosphorothiolated antisense (AS) oligonucleotides to D5, but not D1, dopamine receptor mRNA. Oligonucleotides (2 and 4 nM for D5 and D1 receptors, respectively) were microinjected into indwelling third cerebroventricular cannulae 24 hr after EB priming. Twenty-four hours later, animals were tested for the behavioral effect of EB + oligonucleotide and then challenged with D1-like agonists. Random sense (RS) oligos were given to another group of rats as a control for nonspecific oligo effect.
[View Larger Version of this Image (18K GIF file)]
Specificity for D1 and D5 dopamine receptor oligonucleotides were shown in separate experiments. To ascertain the effect of D1 dopamine receptor mRNA oligos, motor activity was assessed immediately before and 24 hr after intra-accumbal microinjection of antisense (2-4 nM), random sense (4 nM), and/or vehicle as described previously. For all animals receiving antisense to the D1 receptor mRNA, levels of basal motor activity were higher compared with pretreatment levels (Fig. 4 A, lanes 5 vs 4, and 8 vs 7) (ANOVA, p < 0.05). No difference in rearing, sniffing, and grooming was noted. Behavior consistent with amphetamine, serotonin syndrome, or stereotypy was not observed. In random sense-treated and vehicle-treated animals, locomotor activity levels were similar to pretreatment basal activity levels (Fig. 4A, lane 2 vs lane 1). When the persistent effect of D1 receptor oligos was tested after 48 hr, animals pretreated with antisense oligo (4 nM) failed to display additional augmentation of hyperactivity after SKF82958 challenge (Fig. 4A, lane 9 vs lane 8). In contrast, it was only after SKF challenge that hyperactivity was displayed by animals treated with random sense (Fig. 4A, lane 3 vs lane 2).
Fig. 4. Specific effects of antisense (AS) and sense (S) oligonucleotides to D1 (A) and D5 (B) dopamine receptor mRNAs. A, Nuclear accumbens treatment with D1 antisense increases basal motor activity and blocks the induction of additional hyperactivity after SKF82958. A behavioral paradigm known for its D1 dopamine receptor specificity in targeted mutagenic mice (n = 40) was used as in Materials and Methods. Data are replicates of two experiments. B, Treatment with increasing levels of D5 antisense increases the inhibition of D1-like agonist-stimulated CAT gene expression. Mouse L(tk
[View Larger Version of this Image (20K GIF file)]
Agonists for D1-like dopamine receptors activate the PR and ER in a steroid ligand-independent manner in transient transfection assays (Smith et al., 1993
Inhibition of behavioral responses by RU38486 and AS to PRa mRNA
Sexual behavior failed to be facilitated by D1-like agonists in the presence of antisense to PRa mRNA or the antiprogestin RU38486 (Fig. 5). In EB-primed rats, pretreatment with RU38486 suppressed SKF38393-facilitated lordosis (Fig. 5, lane 8). As with RU38486, the effect of SKF38393, SKF75670, or SKF85174 on receptive behavior was seriously attenuated when phosphorothioated antisense to PRa mRNA was given as described above (Fig. 5, lanes 14, 19, 24). In contrast, animals receiving sense oligo exhibited high levels of receptive behavior after intracerebroventricular challenge with SKF38393, SKF75670, or SKF85174 (Fig. 5, lanes 11, 17, 22). In these animals receiving EB-priming and sense or antisense treatment, displays of lordosis and proceptive behaviors were absent 1 hr before the administration of SKF38393 (Fig. 5, lanes 9, 16, 21 for sense, and lanes 12, 18, 23 for antisense). Control animals receiving EB + RU38486 displayed sporadic lordosis responses (Fig. 5, lane 4). As with all oligos, neither vehicle nor RU38486 in the absence of EB was associated with receptive behavior (Fig. 5, lanes 1, 2). Both antisense oligo and RU38486 suppressed the effect of P on sex behavior (Fig. 5, lane 3 vs lanes 5, 13), whereas sense oligo had no effect on P-inducible behavior (Fig. 5, lane 10).
Fig. 5. Inhibition of SKF-induced lordosis response by antisense oligonucleotides to PRa mRNA and the antiprogestin RU38486 (RU486). Rats were administered RU38486 and challenged with D1-like agonist 1 hr later. In another group, oligonucleotides (2.6 nM, i.c.v.) were administered concurrently with EB and 24 hr later. At 48 hr after EB priming, animals received intracerebroventricular SKF challenge and were tested for behaviors as described in Materials and Methods.
[View Larger Version of this Image (21K GIF file)]
Localization of dopaminergic activity and behavior by intranuclear injections
Microinjections of dopaminergic agonists (SKF38393, SKF75640, SKF85174) in the VMN facilitated receptive behavior in EB-primed rats (Fig. 6A, lanes 5, 11, 15). In contrast, 24 hr pretreatment with D5 dopamine receptor mRNA antisense by direct VMN microinjections suppressed SKF-induced receptive behavior (Fig. 6A, lanes 9, 13, 19). As with third ventricle injections, VMN microinjections of sense oligonucleotide had no effect on the induction of receptive behavior when the VMN was challenged with D1-like agonists (Fig. 6A, lanes 7, 12, 16). Both antisense- and sense-treated females failed to display sex behaviors 1 hr before SKF38393 treatment (Fig. 6A, lanes 6, 8, 11, 15). During control experiments, all of these animals responded with LQs above 90% to P challenge (Fig. 6A, lane 3). In another control group of animals with third ventricle cannulas that served to verify the effectiveness of SKF38393 on the day of intranuclear injections, LQs were above 85% (data not shown).
Fig. 6. Facilitation of lordosis by microinjections of D1-like agonists into stereotaxically implanted cannulae in the ventromedial nucleus (VMN) (Panel A), but not arcuate nucleus (AN) (panel B) or preoptic area (POA) (panel B). Fourteen days after screening for false-positive and false-negative behaviors, animals were tested for behavioral responsiveness to SKF38393, SKF75640, or SKF85174. For blockade, D5 antisense oligonucleotide (2 nM total) was injected into VMN cannulae 24 hr before SKF38393 challenge. B presents the responses of those animals with either AN or POA cannulae. Animals served as their own controls for experimental results. Because all responded with LQ above 85%, the data are not shown from another group of animals with third ventricle cannulas that served to verify the effectiveness of D1-like agonists to induce responses on the day of intranuclear injections. Each experiment was repeated two to three times. Cannula placement was verified by microscopic examination at the end of experimentation. Data are shown for those animals with both cannula tips in target area.
[View Larger Version of this Image (23K GIF file)]
For both AN and POA microinjections, SKF38393 failed to induce lordosis responses (Fig. 6B, lanes 21, 25). Excessive displays of aggression were common for females receiving SKF38393 in the POA and were absent before SKF treatment. Aggressive behaviors were less apparent at 4 hr, but vocalizations and rejection remained excessive. Rejection behavior was common when SKF38393 was administered in the AN. Intracerebral nuclei cannulation did not decrease the ability of ovariectomized females to exhibit proceptive or receptive responses when treated with appropriate agonists (Fig. 6B, lanes 20, 22, 23).
Anatomical distribution of D1 and D5 dopamine receptor mRNAs and in situ hybridization
The in situ hybridization studies show that D1 dopamine receptor mRNA are densely distributed throughout the VMN of EB-primed ovariectomized rats as well as the VMN of male rats (Fig. 7D1, A). The area of highest density appeared to be more lateral than medial. In comparison, D5 receptor mRNA was not detected in the VMN of either the EB-primed ovariectomized rat nor the male rat (Fig. 7D5, B). Slides dipped in photoemulsion (NTB2) and exposed for several weeks until background was unacceptably high did not reveal any additional positive cells. As expected from previous studies for distribution of mRNA (Tiberi et al., 1991
Fig. 7. In situ hybridizations and dark-field autoradiograms of D1 (A) and D5 (B) mRNA. Modest distribution of D1, but not D5, dopamine receptor mRNA is visualized in the ventromedial nucleus of the hypothalamus (VMH) 48 hr after estrogen priming of ovariectomized female (E2F in D1A, B; D5A, B) and in the VMH of male (M in D1 C, D; D5C, D) rats. A D1 dopamine receptor cRNA probe transcribed from a rat D1 dopamine receptor cDNA and a D5 dopamine receptor riboprobe generated from a partial rat D5 dopamine receptor cDNA were used. A and C are at low magnification, and B and D are at higher magnification. HIP, Hippocampus; CPu, caudate putamen; 3V, third ventricle; Re, reuniens thalamic nucleus; En, endopiriform nucleus.
[View Larger Version of this Image (73K GIF file)]
DISCUSSION
The present study demonstrates that steroid-dependent behavior can be potentiated by activation of dopaminergic pathways through the membrane-bound D5 dopamine receptor in estrogen-primed, ovariectomized rats. This conclusion is supported by the finding that four D1-like agonists facilitated receptive behavior. In contrast, pretreatment with D1-like antagonist or D1-like/D2 receptor repopulation inhibition blocked behavioral facilitation by D1-like challenge with SKF38393. Importantly, antisense oligonucleotides to D5, but not D1, dopamine receptor mRNA suppressed induction of receptive behavior when animals were challenged with the D1-like agonists SKF38393, SKF75640 ,and SKF85174. Further, potentiation of reproductive behavior by dopaminergic agonists was dependent on PR, because pretreatment with either antiprogestin or PR antisense oligonucleotides also blocked the effect of D1-like agonists. Finally, the effects of D1-like agonists were selective for the VMN, because agonist microinjections into the VMN, but not AR or POA, facilitated induction of proceptivity and receptivity. Further, intranuclear D5 dopamine receptor mRNA antisense and D1-like receptor antagonism or repopulation blockade suppressed facilitation of sex behavior by D1-like agonists. Collectively, the data provide strong evidence for dopaminergic modulation of reproductive behavior through D5 dopamine receptor-linked second messenger pathways in a PR-dependent manner.The present results support previous studies (Everitt et al., 1975
The functional and biochemical consequences of D1-like heterogeneity are difficult to assess, because no subtype-selective agonists or antagonists yet exist. However, antisense oligonucleotides for sequence-specific inhibition of gene expression are effective for studying the biological effects of specific proteins (Cohen, 1989
Using in situ hybridization, mRNAs for D1, but not D5, dopamine receptors are demonstrated in the VMN of EB-primed ovariectomized rats in the present study, a finding consistent with previous studies of male rats (Mansour et al., 1992
Finally, the present study confirms the dependence of dopamine-facilitated reproductive behavior on the PR. Regardless of D1-like agonists, the dopaminergic facilitatory effect was blocked by the antiprogestin RU38486 and antisense to PR. The site of influence was determined by direct microinjections of D1-like agonists and antisense oligos into the VMN, AN, and POA. Because the site of dopaminergic influence of PR-mediated events is within the VMN, it is possible that D5 dopamine receptor-mediated stimulation converted PR to an active form within individual neurons through the alternate ligand-independent pathway shown in vitro for PR (and ER) (Tsai and O'Malley, 1994
Alternately, modulation of the lordosis reflex could be attributable to local interneuron circuitry. Execution of lordosis is dependent on VMN projections to periaqueductal gray and subject to modulation by neurons in the POA and AN (Pfaff and Schwartz-Giblin, 1988
In conclusion, four D1-like agonists mimic P and facilitate induction of reproductive behavior in EB-primed ovariectomized rats. The D1-like antagonist SCH23390 and, specifically, D5, but not D1, antisense attenuate facilitation of lordosis by D1-like agonists. Dopaminergic stimulation of the VMN, but not AN or POA, was critical for regulation of estrogen-dependent behavior. Although not detectable by in situ hybridization, the ability of VMN microinjections of D5 antisense block the induction of reproductive behavior by D1-like agonists suggests the functional presence of D5 dopamine receptors in the VMN. The data provide strong evidence for PR-dependence of dopamine-induced lordosis, because pretreatment with either PR antisense or antiprogestin RU38486 blocked the effect of D1-like agonists. Taken together, the data support the hypothesis that activation of dopaminergic pathways can modulate reproductive behavior by activation of the PRs in the absence of its cognate ligand.
FOOTNOTES
Received Dec. 27, 1995; revised May 8, 1996; accepted May 13, 1996.
This work was supported by National Institutes of Health Grant HD-07857 (B.O.M.), National Institutes of Mental Health Grant P01 MH42251 (S.J.W.), and National Scientist Research Award Grant NRO6826 (E.M.A.). We thank K. E. Mayo (Northwestern University) for providing rat mRNA sequence, M. Caron (Duke University) for providing D5-expressing cell line, W. W. Vale (The Salk Institute) for providing GnRH, and SmithKline & Beecham for providing SKF75670.
Preliminary results of this study have been published as an abstract at the Annual Meeting of the Endocrinology Society, June 15-19, 1994 (Anaheim, CA).
Correspondence should be addressed to Dr. Bert W. O'Malley, Department of Cell Biology, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030.
REFERENCES
- Andersen PH, Jansen JA (1990) Dopamine receptor agonists: selectivity and dopamine D1 receptor efficacy. Eur J Pharmacol Mol Pharmacol Sect 188:335-347 . [ISI][Medline]
- Boiziau C, Thuong NT, Toulme J-J (1992) Mechanisms of the inhibition of reverse transcription by antisense oligonucleotides. Proc Natl Acad Sci USA 89:768-772 .
[Abstract/Free Full Text] - Cauteras NS, Simerly RB, Swanson LW (1994) Organization of projections from the ventromedial nucleus of the hypothalamus: a phaseolus vulgaris-Leucoagglutinin study in the rat. J Comp Neurol 348:41-79. [ISI][Medline]
- Cohen JS (1989) Oligodeoxyribonucleotides. Antisense inhibitors of gene expression. In: Topics in molecular and structural biology, (Cohen, JS, eds) , Vol 12, p. 1. London: Macmillan.
- Cristiano RJ, Smith LC, Woo SLC (1993) Hepatic gene therapy: adenovirus enhancement of receptor-mediated gene delivery and expression in primary hepatocytes. Proc Natl Acad Sci USA 90:2122-2126 .
[Abstract/Free Full Text] - Dluzen DE, Ramirez VD (1989a) Progesterone effects upon dopamine release from the corpus striatum of female rats. I. Evidence for a membrane-bound progesterone receptor. Brain Res 476:332-337 . [ISI][Medline]
- Dluzen DE, Ramirez VD (1989b) Progesterone effects upon dopamine release from the corpus striatum of female rats. II. Evidence for a membrane site of action and the role of albumin. Brain Res 476:338-344 . [ISI][Medline]
- Dreher JK, Jackson DM (1989) Role of D1 and D2 dopamine receptors in mediating locomotor activity elicited from the nucleus accumbens of rats. Brain Res 487:267-277 . [ISI][Medline]
- Ehlrich G, Patinkin D, Ginzberg D, Zakut H, Eckstein F, Soreq H (1994) Use of partially phosphorothioated ``antisense'' oligodeoxynucleotides for sequence-dependent modulation of hematopoiesis in culture. Antisense Res Dev 4:173-183. [ISI][Medline]
- Etgin AA (1990) Intrahypothalamic implants of noradrenergic antagonists disrupt lordosis behavior in female rats. Physiol Behav 48:31-36. [Medline]
- Evans RM (1988) The steroid and thyroid hormone receptor superfamily. Science 240:889-895 .
[Abstract/Free Full Text] - Everitt BJ, Fuxe K (1977) Dopamine and sexual behaviour in female rats. Effect of dopamine receptor agonists and sulpride. Neurosci Lett 4:209-213.
- Everitt BJ, Fuxe K, Hokfelt T, Jonsson G (1975) Pharmalogical and biochemical studies on the role of monoamines in the control by hormones of sexual receptivity in the female rat. J Comp Physiol Psychol 89:556-572 . [ISI][Medline]
- Foreman M, Moss R (1979) Role of hypothalamic dopaminergic receptors in the control of lordosis behavior in the female rat. Physiol Behav 22:283-289 . [Medline]
- Fox CA, Mansour A, Thompson RC, Bunzow JR, Civelli O, Watson SJ (1993) The distribution of dopamine D2 receptor heteronuclear RNA (hnRNA) in the rat brain. J Chem Neuroanat 6:363-373 . [ISI][Medline]
- Fox CA, Mansour A, Watson SJ (1994) The effects of haloperidol on dopamine receptor gene expression. Exp Neurol 130:1-16. [ISI][Medline]
- Grierson JP, James MD, Pearson JR, Wilson CA (1988) The effect of selective D1 and D2 dopaminergic agents on sexual receptivity in the female rat. Neuropharmacology 27:181-189 . [ISI][Medline]
- Kow L-M, Mobbs CV, Pfaff DW (1994) Roles of second-messenger systems and neuronal activity in the regulation of lordosis by neurotransmitters, neuropeptides, and estrogen: a review. Neurosci Biobehav Rev 18:251-268 . [ISI][Medline]
- Mahan LC, Burch RM, Monsma FJ Jr, Sibley DR (1990) Expression of striatal D1 dopamine receptors coupled to inositol phosphate production and Ca2+ mobilization in Xenopus oocytes. Proc Natl Acad Sci USA 87:2196-2200 .
[Abstract/Free Full Text] - Mani SK, Allen JMC, Blaustein Clark JH, O'Malley BW (1994) Convergent pathways for steroid hormone- and neurotransmitter-induced rat sexual behavior. Science 265:1246-1249 .
[Abstract/Free Full Text] - Mansour A, Meador-Woodruff JH, Bunzow JR, Civelli O, Akil H, Watson SJ (1990) Localization of dopamine D2 receptor mRNA and D1 and D2 receptor binding in the rat brain and pituitary: an in situ hybridization-receptor autoradiographic analysis. J Neurosci 10:2587-2600 . [Abstract]
- Mansour A, Meador-Woodruff JH, Zhou Q, Civelli O, Akil H, Watson SJ (1992) A comparison of D1 receptor binding and mRNA in rat brain using receptor autoradiographic and in situ hybridization techniques. Neuroscience 46:959-971 . [ISI][Medline]
- McCarthy MM, Brooks PJ, Pfaus JG, Brown HE, Flanagan LM, Schwartz-Giblin S, Pfaff DW (1993) Antisense oligonucleotides in behavioral neuroscience. Neuroprotocols 2:67-74.
- Meador-Woodruff JH, Mansour A, Grandy DK, Damask SP, Civelli O, Watson SJ (1992) Distribution of D5 dopamine receptor mRNA in rat brain. Neurosci Lett 145:209-212 . [ISI][Medline]
- O'Boyle KM, Gaitanopoulos DE, Brenner M, Waddington JL (1989) Agonist and antagonist properties of benzazepine and thienopyridine derivatives at the D1 dopamine receptor. Neuropharmacology 28:401-405. [ISI][Medline]
- Paxinos G, Watson C (1988) The rat brain in stereotaxic coordinates. .
- Pfaff DW, Schwartz-Giblin S (1988) Cellular mechanisms of female reproductive behaviors. In: The physiology of reproduction, (Knobil, E, Neill, JD, Ewing, LL, Greenwald, GS, Markert, CL, Pfaff, DW, eds) , Vol 2, p. 1487. New York: Raven.
- Phillips LL, Nostrandt SJ, Chikaraishi DM, Steward O (1987) Increases in ribosomal RNA within the denervated neurophil of the dentate gyrus during reinnervation: evaluation by in situ hybridization using DNA probes complementary to ribosomal RNA. Mol Brain Res 2:251-261.
- Pollio G, Xue P, Zanisi M, Nicolin A, Maggi A (1993) Antisense oligonucleotide blocks progesterone-induced lordosis behavior in ovariectomized rats. Mol Brain Res 19:135-139 . [Medline]
- Rao A, Steward O (1991) Evidence that protein constituents of postsynaptic membrane specializations are locally synthesized: analysis of proteins synthesized within synaptosomes. J Neurosci 11:2881-2895 . [Abstract]
- Smith CL, Conneely OM, O'Malley BW (1993) Modulation of the ligand-independent activation of the human estrogen receptor by hormone and antihormone. Proc Natl Acad Sci USA 90:6120-6124 .
[Abstract/Free Full Text] - Steward O, Banker GA (1992) Getting the message from the gene to the synapse: sorting and intracellular transport of RNA in neurons. Trends Neurosci 15:180-185 . [ISI][Medline]
- Strong MJ, Svedmyr A, Gajdusek DC, Garruto RM (1990) The temporal expression of amyloid precursor protein mRNA in vitro in dissociated hippocampal neuron cultures. Exp Neurol 109:171-179 . [ISI][Medline]
- Tiberi M, Jarvio KR, Slivia C, Falardeau P, Gingrich JA, Godinot N, Bertrand L, Yang-Feng TL, Fremeau RT Jr, Caron MG (1991) Cloning, molecular characterization, and chromosomal assignment of a gene encoding a second D-1 dopamine receptor subtype: differential expression pattern in rat brain compared with the D1
receptor. Proc Natl Acad Sci USA 88:7491-7495 .
[Abstract/Free Full Text] - Tsai M-J, O'Malley BW (1994) Molecular mechanisms of action of steroid/thyroid receptor superfamily members. Annu Rev Biochem 63:451-486 . [ISI][Medline]
- Undie AS, Friedman E (1992) Selective dopaminergic mechanism of dopamine and SKF38393 stimulation of inositol phosphate formation in rat brain. Eur J Pharmacol Mol Pharmacol Sect 226:297-302 . [ISI][Medline]
- Xu M, Morstall R, Gold LH, Hirol N, Koob GF, Graybiel Tonegawa S (1994) Dopamine D1 receptor mutant mice are deficient in striatal expression of dynorphin and in dopamine-mediated behavioral responses. Cell 79:729-742 . [ISI][Medline]
- Zhou Q-Y, Grandy DK, Thambi L, Kushner JA, Van Tol HHM, Cone R, Pribhow D, Salon J, Bunzow JR, Civelli O (1990) Cloning and expression of human and rat D1 dopamine receptors. Nature 347:76-79 . [Medline]
Copyright ©1996 Society for Neuroscience 0270-6474/1996/164823-12$05.00/0
This article has been cited by other articles:
N. Vasudevan and D. W. Pfaff
Membrane-Initiated Actions of Estrogens in Neuroendocrinology: Emerging Principles
Endocr. Rev., February 1, 2007; 28(1): 1 - 19.
[Abstract] [Full Text] [PDF]
S. Y. Kim, K. C. Choi, M. S. Chang, M. H. Kim, S. Y. Kim, Y.-S. Na, J. E. Lee, B. K. Jin, B.-H. Lee, and J.-H. Baik
The dopamine D2 receptor regulates the development of dopaminergic neurons via extracellular signal-regulated kinase and Nurr1 activation.
J. Neurosci., April 26, 2006; 26(17): 4567 - 4576.
[Abstract] [Full Text] [PDF]
J. D. Blaustein
Minireview: Neuronal Steroid Hormone Receptors: They're Not Just for Hormones Anymore
Endocrinology, March 1, 2004; 145(3): 1075 - 1081.
[Abstract] [Full Text] [PDF]
T. R. Hollon, M. J. Bek, J. E. Lachowicz, M. A. Ariano, E. Mezey, R. Ramachandran, S. R. Wersinger, P. Soares-da-Silva, Z. F. Liu, A. Grinberg, et al.
Mice Lacking D5 Dopamine Receptors Have Increased Sympathetic Tone and Are Hypertensive
J. Neurosci., December 15, 2002; 22(24): 10801 - 10810.
[Abstract] [Full Text] [PDF]
N. Vasudevan, L.-M. Kow, and D. W. Pfaff
Early membrane estrogenic effects required for full expression of slower genomic actions in a nerve cell line
PNAS, September 19, 2001; (2001) 221449798.
[Abstract] [Full Text] [PDF]
S. K. Mani, A. A. Fienberg, J. P. O'Callaghan, G. L. Snyder, P. B. Allen, P. K. Dash, A. N. Moore, A. J. Mitchell, J. Bibb, P. Greengard, et al.
Requirement for DARPP-32 in Progesterone-Facilitated Sexual Receptivity in Female Rats and Mice
Science, February 11, 2000; 287(5455): 1053 - 1056.
[Abstract] [Full Text]
J. M. Meredith, C. A. Moffatt, A. P. Auger, G. L. Snyder, P. Greengard, and J. D. Blaustein
Mating-Related Stimulation Induces Phosphorylation of Dopamine- and Cyclic AMP-Regulated Phosphoprotein-32 in Progestin Receptor-Containing Areas in the Female Rat Brain
J. Neurosci., December 1, 1998; 18(23): 10189 - 10195.
[Abstract] [Full Text] [PDF]
S.-H. Yen and J.-T. Pan
Progesterone Advances the Diurnal Rhythm of Tuberoinfundibular Dopaminergic Neuronal Activity and the Prolactin Surge in Ovariectomized, Estrogen-Primed Rats and in Intact Proestrous Rats
Endocrinology, April 1, 1998; 139(4): 1602 - 1609.
[Abstract] [Full Text] [PDF]
M. R. Walters, M. Dutertre, and C. L. Smith
SKF-82958 Is a Subtype-selective Estrogen Receptor-alpha (ERalpha ) Agonist That Induces Functional Interactions between ERalpha and AP-1
J. Biol. Chem., January 11, 2002; 277(3): 1669 - 1679.
[Abstract] [Full Text] [PDF]
N. Vasudevan, L.-M. Kow, and D. W. Pfaff
Early membrane estrogenic effects required for full expression of slower genomic actions in a nerve cell line
PNAS, October 9, 2001; 98(21): 12267 - 12271.
[Abstract] [Full Text] [PDF]
This Article Abstract 
Full Text (PDF) Submit an eLetter Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal
ABSTRACT
