Alpha-1- and alpha-2-noradrenergic receptors modulate lordosis behavior in female guinea pigs

Modulation of lordosis behavior by stimulation of noradrenergic receptor subtypes was examined in ovariectomized, estradiol benzoate (10 micrograms)-primed guinea pigs. In the first experiment, subcutaneous administration of the alpha-2-noradrenergic agonist, UK-14,304 (0.01, 0.5, or 1.0 mg/kg, Pfizer Central Research), produced a significant increase in lordosis behavior when these animals were compared with estrogen-primed control animals injected with saline. In a second experiment, estrogen-primed animals were injected with the alpha-1-noradrenergic agonist methoxamine (0.5 mg/kg, s.c.), UK-14,304 (0.5 mg/kg, s.c.), or both drugs given together. Methoxamine or UK-14,304 administered alone facilitated lordosis in fewer than 50% of animals (17 and 39%, respectively). However, when both drugs were given together, 76% of the animals became sexually receptive. A third experiment showed that lordosis behavior facilitated by UK-14,304 could be attenuated by the administration of the alpha-2-noradrenergic antagonist idazoxan (2.5 mg/kg, s.c.). Only 29% of sexually receptive animals (i.e. those animals primed with estradiol benzoate plus UK-14,304) continued to show lordosis after they received idazoxan. The results obtained from these and previously reported experiments suggest that both alpha-1- and alpha-2-noradrenergic receptors are involved in the regulation of lordosis behavior in the guinea pig.     

Participation of growth factor signal transduction pathways in estradiol facilitation of female reproductive behavior

Estradiol (E(2)) regulates female reproductive behavior (lordosis) by acting on estrogen-sensitive neurons. We recently showed that E(2) facilitation of lordosis behavior requires concurrent activation of brain IGF-I receptors. The present study confirmed this finding and sought to identify the downstream signaling pathways involved in estrogen/IGF-I priming of lordosis. Intracerebroventricular infusions of a selective IGF-I receptor antagonist were administered to ovariectomized rats every 12 h beginning 1 h before the first of two daily E(2) injections. IGF-I receptor blockade partially inhibits lordosis if the antagonist is infused throughout the 2-d estrogen treatment period but not if it is administered only during the first or last 12 h of estrogen treatment. Because E(2) and IGF-I can activate phosphatidylinositol-3-kinase (PI3K) and MAPK, we infused agents that block PI3K and/or MAPK activity as described above. Both PI3K inhibitors (wortmannin and LY294002) and MAPK inhibitors (PD98059 and U0126) partially attenuate lordosis when administered during estrogen priming. None of these drugs modifies lordosis if they are infused only once, during the last 12 h of estrogen treatment. When both wortmannin and PD98059 are infused during E(2) priming, lordosis behavior is completely abolished. These data suggest that activation of both PI3K and MAPK by E(2) and IGF-I mediates hormonal facilitation of lordosis behavior.     

The membrane actions of estrogens can potentiate their lordosis behavior-facilitating genomic actions

The membrane actions of estrogens can facilitate their genomic actions. To determine whether this facilitation bears on CNS mechanisms for estrogen-dependent behaviors, ovariectomized rats were subjected to a two-pulse treatment of estrogen directly in the hypothalamic ventromedial nucleus. Two days later, each rat was given progesterone and then tested for lordosis behavior, the induction of which requires the genomic actions of estrogen. When estrogen was given in both pulses (15 min to 2 h duration, and 5 h apart) lordosis was induced. Based on results from studies on neuroblastoma cells, we hypothesized that the membrane actions of estrogen in the first pulse would potentiate the genomic actions of estrogen in the second. This hypothesis was confirmed with the use of a membrane-impermeable estrogen. However, surprisingly, the order of the pulses could be reversed and still achieve lordosis behavior induction. Finally, activators of protein kinase A or PKC were effective substitutes for the membrane-limited pulse of estrogen. Thus, estrogen-induced membrane actions in the hypothalamus can potentiate its lordosis-inducing genomic actions on behavior and may be mediated by signaling pathways involving the activation of protein kinase A and PKC.     

Diurnal rhythmicity of beta-1- and beta-2-adrenergic receptors in ovariectomized, ovariectomized estradiol-treated and proestrous rats

The effect of norepinephrine on LH is complex: the ovarian steroidal milieu appears to determine whether norepinephrine stimulates or inhibits LH secretion. It has been proposed that steroids allow norepinephrine to have opposite effects on LH by altering the relative concentrations of alpha 1-(stimulatory) and beta-(inhibitory) adrenergic receptors in the hypothalamus. Thus, many investigators have argued that estradiol may permit norepinephrine to stimulate LH release by increasing the density of alpha 1- and decreasing the density of beta-adrenergic receptors in one or more key hypothalamic regions. To test this hypothesis specifically, we measured the density of beta 1- and beta 2-adrenergic receptor densities in proestrous, ovariectomized, and ovariectomized estradiol-treated rats at various times of day to determine (1) whether the densities of beta-receptors exhibit diurnal rhythmicity, (2) whether beta-receptors decrease during the time of increased LH secretion and/or (3) how steroidal milieu influences the density and/or the rhythm of receptor densities. The densities of beta 1- and beta 2-receptors exhibit a diurnal rhythm in some brain areas. These rhythms are detectable only in proestrous and ovariectomized rats and only in selected brain regions. Estrogen treatment has opposing effects in different brain regions. It suppresses the rhythm of beta 1-receptor concentrations in ovariectomized rats and also suppresses the average density of receptors in the suprachiasmatic nucleus and pineal gland. In contrast, estrogen increases the density of beta 1-receptors in the medial preoptic nucleus.(ABSTRACT TRUNCATED AT 250 WORDS)     

Estrogen modulation of mu-opioid receptor-stimulated [35S]-GTP-gamma-S binding in female rat brain visualized by in vitro autoradiography

The mu-opioid receptor (OR) is involved in several aspects of female reproductive neuroendocrinology, such as the control of gonadotropin release and the display of lordosis behavior. Even though the neuroendocrine events modulated by mu-ORs are steroid hormone-dependent, few studies have shown how steroid hormones such as estrogen and/or progesterone can affect mu-OR function. Therefore, the present study investigated if in vivo estrogen or estrogen plus progesterone treatment of ovariectomized (OVX) rats affects mu-OR coupling to its G proteins. We used autoradiographic analysis of agonist-stimulated [(35)S]-GTPgammaS binding, in which brain sections were incubated in the presence or absence of the mu-OR agonist [D-Ala(2), N-Me-Phe(4), Gly(2)ol]-enkephalin (DAMGO). Film images were quantified using calibrated [(14)C] standards. Analysis was performed in steroid-responsive hypothalamic regions such as the medial preoptic area (mPOA) and the ventromedial nucleus of the hypothalamus, as well as in non-hypothalamic brain regions. Treatment with estrogen, alone or with progesterone, significantly increased DAMGO-stimulated [(35)S]-GTPgammaS binding in the mPOA when compared to control OVX animals. In addition, estrogen increased mu-OR coupling in the caudate putamen. Steroid treatment had no effect on either basal or DAMGO-stimulated binding in the other brain regions examined. These findings suggest that estrogen modulates mu-OR function in a brain region-specific fashion. This could have important implications in terms of how these hormones synchronize reproductive behavior and gonadotropin release.     

Effects of neuropeptide Y (NPY) and NPY agonists on lordosis in the female guinea pig

We have previously shown that an NPY antagonist decreases lordosis behavior and that this decrease can be reversed with NPY administration. The present experiments examined whether intracerebroventricular (icv) administration of NPY would facilitate lordosis behavior and whether it would increase feeding behavior in the female guinea pig. Additionally, we examined whether icv administration of a more specific NPY Y1 and/or Y2 receptor agonist would facilitate lordosis behavior. Although NPY (1 μg) increased feeding behavior when it was administered to the lateral ventricle of ovariectomized (ovx) estrogen (i.e., estradiol benzoate; EB) and progesterone- (P) treated guinea pigs, it had no facilitatory effect on lordosis behavior at any of the doses tested (0.5, 1, 5, or 10μg). In fact, the lower doses had a small, delayed inhibitory effect. NPY also had no effect on lordosis in females treated with EB alone. In contrast, the NPY Y1 agonist (Leu³¹Pro³⁴) NPY significantly facilitated lordosis in ovx EB- and P-treated females. It had no effect in ovx females treated with EB alone. The NPY Y2 agonist NPY (13–36) had a slight, delayed inhibitory effect in ovx EB- and P-treated females. These data are consistent with the hypothesis that NPY can act at a number of receptor subtypes to affect lordosis behavior, and that NPY can facilitate lordosis behavior by acting at Y1 receptors. Furthermore, it appears that this facilitatory effect of Y1 receptors is an effect on some progesterone-mediated component of lordosis, as the Y1 agonist facilitated EB- and P-induced lordosis, but not that induced with EB alone.     

The neuropeptide Y antagonist PYX2 decreases lordosis behavior

Neuropeptide Y (NPY) has been localized to noradrenergic neurons and both the noradrenergic system and NPY play a facilitatory role in the control of luteinizing hormone-releasing hormone (LHRH) and luteinizing hormone (LH) release. The present experiments examined whether NPY also plays a role in the control of lordosis. Adult female guinea pigs were ovariectomized (ovx) and implanted with a cannula into the lateral ventricle. In Experiment 1, intracerebroventricular (ICV) administration of the NPY antagonist PYX2 (0, 0.5, 2.0 or 10.0 μg) caused a dose-dependent decrease in lordosis behavior in ovx, estrogen and progesterone-primed guinea pigs. In addition to an effect on the mean lordosis response, PYX2 also decreased the percent of animals showing lordosis and the maximum lordosis response. In Experiment 2, NPY administration (25 μg, ICV) 30 min after PYX2 (2 μg, ICV) to ovx estrogen and progesterone-primed females significantly reversed the effect of the PYX2. Because the NPY antagonist PYX2 reversibly decreased lordosis behavior this suggests that NPY plays a facilitatory role in the control of lordosis behavior.     

GABAergic control of receptivity in the female rat

GABAergic neurotransmission has been implicated in the control of the steroid-dependent behavior, lordosis. GABA has dual effects on lordosis: it facilitates lordosis through actions in the medial hypothalamus (mHYP) and it inhibits lordosis through actions in the preoptic area (POA). In the present study, gonadally intact and ovariectomized female rats were behaviorally tested with a sexually active male. Brains were removed from sexually receptive female either 1 or 24 h after behavioral testing. There was a significant difference in endogenous GABA concentration in HYP and POA between receptive, postreceptive and ovariectomized nonreceptive females. Specifically, GABA levels in postreceptive females were higher in the HYP (20%) and lower in the POA (21%) in comparison to receptive females (p less than 0.05). There was also a significant change in binding parameters of 3H-muscimol in the HYP and POA of receptive females as compared to 24 h postreceptive and ovariectomized rats. Attempts to modulate 3H-GABA release from hypothalamic tissue slices by estrogen or progesterone in ovariectomized rats yielded no effect on this parameter.     

In the ventral tegmental area, G-proteins and cAMP mediate the neurosteroid 3alpha,5alpha-THP's actions at dopamine type 1 receptors for lordosis of rats

Progestins have multiple mechanisms of action in the central nervous system that are important for modulating lordosis of female rats. In the ventral tegmental area (VTA), progestins, such as the progesterone metabolite and neurosteroid 5alpha-pregnan-3alpha-ol-20-one (3alpha,5alpha-THP), regulate lordosis via actions independent of intracellular progestin receptors. We hypothesized that if, in the VTA, dopamine type 1 receptors (D1), G-proteins, and adenosine 3',5'-monophosphate (cAMP) are downstream effectors of 3alpha,5alpha-THP's actions for lordosis, then pharmacological manipulations of these signaling molecules will produce changes in 3alpha,5alpha-THP-facilitated lordosis of estradiol (E2)-primed rats. VTA infusions of 3alpha,5alpha-THP (50 ng) or 3alpha,5alpha-THP and the D1 agonist SKF38393 (100 ng) increased lordosis of ovariectomized, E2 (10 microg)-primed rats, compared to vehicle. Both 3alpha,5alpha-THP- and 3alpha,5alpha-THP plus SKF38393-facilitated lordosis was reduced by VTA infusions of the G-protein inhibitor guanosine 5'-O-(2-thiodiphosphate) (GDP-beta-S; 50 microM), but not vehicle. Also, in the VTA, blocking D1 with SCH23390 (100 ng) decreased, or increasing cAMP with 8-bromo-cAMP (200 ng) enhanced, 3alpha,5alpha-THP-facilitated lordosis of E2-primed rats. Notably, SCH23390's inhibitory effects on 3alpha,5alpha-THP-facilitated lordosis were reversed by 8-bromo-cAMP. Thus, in the VTA, 3alpha,5alpha-THP's actions for lordosis may involve activation of D1 and initiation of the G-protein-mediated second messenger cAMP.     

Regulation of lordosis by cyclic 3',5'-guanosine monophosphate, progesterone, and its 5alpha-reduced metabolites involves mitogen-activated protein kinase

Progesterone (P) and its ring A-reduced metabolites regulate sexual behavior in ovariectomized, estrogen-primed female rats when they are administered intracerebrally and systemically. The present study tested the hypothesis that the MAPK pathway participates in P facilitation and sequential inhibition of sexual behavior. The role of MAPK in lordosis facilitation by two ring A-reduced metabolites of P, 5alpha-dihydroprogesterone (5alpha-DHP) and 5alpha,3alpha-pregnanolone (5alpha,3alpha-Pgl), was also assessed. In Experiment 1, the MAPK inhibitor PD98059 was infused intracerebroventricularly before progestin administration. Lordosis behavior induced by P, 5alpha-DHP, and 5alpha,3alpha-Pgl was abolished 2 h after progestin administration by PD98059. P and 5alpha,3alpha-Pgl facilitation of proceptive behaviors was also decreased by the MAPK inhibitor. Experiment 2 examined the effects of MAPK inhibition on P sequential inhibition. Estrogen-primed females received intracerebroventricular infusions of PD98059 or vehicle 30 min before systemic administration of P and were tested for lordosis 4 h later. Animals received a second injection of P 24 h later and were retested for lordosis. The MAPK inhibitor blocked both lordosis facilitation and sequential inhibition produced by systemic administration of P. Because cGMP can also facilitate lordosis behavior, and cGMP-dependent protein kinase can activate MAPK, experiment 3 determined whether interference with MAPK would affect cGMP enhancement of lordosis. The icv infusion of PD98059 significantly inhibited lordosis behavior induced by 8-bromo-cGMP, a cell-permeable cGMP analog, at both 2 and 4 h. These data support the hypothesis that the MAPK pathway is involved in lordosis regulation by P and some of its ring A-reduced metabolites as well as by the second messenger, cGMP.     

Estrogen alters the diurnal rhythm of alpha 1-adrenergic receptor densities in selected brain regions

Norepinephrine regulates the proestrous and estradiol-induced LH surge by binding to alpha 1-adrenergic receptors. The density of alpha 1-receptors may be regulated by estradiol, photoperiod, and noradrenergic neuronal activity. We wished to determine whether alpha 1-receptors exhibit a diurnal rhythm in ovariectomized and/or estradiol-treated female rats, whether estradiol regulates alpha 1-receptors in those areas of brain involved with LH secretion and/or sexual behavior, and whether the concentrations of alpha-receptors vary inversely relative to previously reported norepinephrine turnover patterns. Young female rats, maintained on a 14:10 light-dark cycle were ovariectomized. One week later, half of them were outfitted sc with Silastic capsules containing estradiol. Groups of animals were decapitated 2 days later at 0300, 1000, 1300, 1500, 1800, and 2300 h. Brains were removed, frozen, and sectioned at 20 micron. Sections were incubated with [3H]prazosin in Tris-HCl buffer, washed, dried, and exposed to LKB Ultrofilm. The densities of alpha 1-receptors were quantitated using a computerized image analysis system. In ovariectomized rats, the density of alpha 1-receptors exhibited a diurnal rhythm in the suprachiasmatic nucleus (SCN), medial preoptic nucleus (MPN), and pineal gland. In SCN and MPN, receptor concentrations were lowest during the middle of the day and rose to peak levels at 1800 h. In the pineal gland, the density of alpha 1-receptors was lowest at middark phase, rose to peak levels before lights on, and remained elevated during the day. Estradiol suppressed the density of alpha 1 binding sites in the SCN, MPN, median eminence, ventromedial nucleus, and the pineal gland but had no effect on the lateral septum. Estrogen treatment altered the rhythm of receptor densities in MPN, median eminence, and the pineal gland. In MPN, a transient suppression in the density of receptors occurred at 1500 h, a time associated with increased norepinephrine turnover and the onset of the LH surge. Therefore, we conclude that alpha 1-receptor densities undergo a diurnal rhythm in brain regions associated with entrainment to the photoperiod. Estrogen alters the rhythm of alpha 1-receptors in areas involved with the regulation of LH secretion and decreases the density in other estrogen-responsive regions.     

Alpha 2-adrenergic receptors are involved in the suppression of luteinizing hormone release during acute fasting in the ovariectomized estradiol-primed rats.

It has been previously reported that the adrenergic system is involved in the control of feeding behavior and LH release. In the present study, the role of the adrenergic receptors in the suppression of LH release during acute fasting are examined by injecting the alpha 1-antagonist (prazosin), alpha 2-antagonists (idazoxan, SKF 86466-A, piperoxan), or beta-antagonist (propranolol) into the third ventricle of unfasted and 48 h fasted ovariectomized estradiol-treated rats. Blood samples were collected every 6 min for 3 h and the drugs were administered after the first hour of the sampling period. Prazosin caused a significant suppression of LH release in the unfasted animals while idazoxan and propranolol had no significant effects. In contrast, all alpha 2-antagonists blocked the inhibitory effect of fasting on LH release and significantly reinstated the suppressed LH release while prazosin and propranolol had no significant effects. We conclude from these results that the suppression of LH release during acute fasting is mediated by alpha 2-adrenergic receptors but not alpha 1- or beta-adrenergic receptors.     

Inhibition of estrous behavior by progesterone in rats: role of neural estrogen and progestin receptors

Sexual receptivity can be induced in ovariectomized rats with sequential injections of 17 beta-estradiol-3-benzoate (EB; 2 micrograms sc) and progesterone given 24 h apart. If a high dose of progesterone is used, estrous behavior is followed by a period during which sexual receptivity cannot be reinduced with a second progesterone injection (sequential inhibition). We have examined the effects of a hormone treatment which causes a sequential inhibition on levels of estrogen and progestin receptors in the hypothalamus-preoptic area-septum of ovariectomized rats. In a preliminary experiment, a single injection of 2 micrograms EB caused a significant increase in the concentration of brain cell nuclear estrogen receptors, Peak levels were seen at 12-24 h and remained significantly elevated as long as 72 h after EB. In a second experiment, we found that 48 h after EB, brain cytoplasmic estrogen receptors were significantly depressed and nuclear estrogen receptors were significantly elevated. However, a single progesterone injection (5 mg, sc) 24 h after EB had no significant effect on either cytoplasmic or nuclear estrogen receptor levels. Forty-eight hours after EB, brain cytoplasmic progestin receptors were significantly elevated compared with oil-injected controls. In a third experiment, progesterone (5 mg, sc) given 24 h after EB reduced neural cytoplasmic progestin receptors to levels significantly below those of oil-injected controls at 48 h. These results are consistent with the suggestion that progesterone does not interact with neural estrogen receptors to inhibit lordosis in rats. Rather, we suggest that progesterone may induce a behavioral refractoriness to subsequent progesterone treatments, at least in part, by reducing the concentration of neural cytoplasmic progestin receptors.     

GABAergic regulation of lordosis: influence of gonadal hormones on turnover of GABA and interaction of GABA with 5-HT.

The role of GABAergic neurons in activating female sexual behavior and possible mechanisms for GABAergic effects on behavior were examined in female rats. First, effects of the ovarian hormones estrogen and progesterone (P), at doses which promote lordosis, on levels and turnover/activity of GABA, were examined in brain areas which regulate lordosis. Utilizing AOAA, an inhibitor of GABA degradation, the accumulation rate of GABA (turnover/activity) was assessed in ovariectomized (Ovx), Ovx + estrogen and Ovx + estrogen + P-treated rats. Estradiol increased GABA accumulation rates in the arcuate-median eminence and in the area dorsal to and surrounding the VMN (VMN-S). P administration following estrogen priming enhanced GABA turnover in the medial preoptic area (mPOA) and further increased turnover in the VMN-S while GABA turnover decreased in the dorsomedial nucleus. No effects of hormones were noted in the VMN itself or in the dorsal midbrain central gray. Reverse dialysis of the GABAA antagonist bicuculline into the basomedial hypothalamus was associated with a time-dependent inhibition of lordosis and a 300% increase in 5-HT release in the basomedial hypothalamus as measured by in vivo dialysis. These results provide additional evidence that GABAergic neurons mediate the physiological regulation of female sexual behavior and suggest that such mediation may involve an interaction with 5-HT containing neurons.     

Effects of adrenergic blockers on central nervous system-mediated hyperglycemia in fed rats.

We studied the effect of adrenergic blockade on hepatic venous hyperglycemia and the activation of a hepatic glycogenolytic enzyme, phosphorylase-a, in response to cerebral cholinergic activation. Neostigmine was injected into the third cerebral ventricle of bilaterally adrenodemedullectomized (ADMX) rats, while somatostatin and insulin were administered intravenously. Hepatic venous plasma glucose concentrations and hepatic phosphorylase-a activity were measured. Intracerebroventricular injection of neostigmine (5 x 10(-8) mol) caused increases in hepatic venous glucose concentrations and hepatic phosphorylase-a activity. Both of these changes were prevented by intraperitoneal (IB) pretreatment with phentolamine (5 x 10(-7), 1 x 10(-6) mol) without the intervention of insulin secretion, but not by pretreatment with the alpha-adrenoreceptor antagonist phenoxybenzamine (1 x 10(-6) mol), the beta-adrenoreceptor antagonist propranolol (1 x 10(-6) mol), the alpha 1-antagonists prazosin or bunazosin (1 x 10(-6) mol), the alpha 2-antagonist yohimbine (1 x 10(-6) mol), or prazosin (5 x 10(-7) mol) plus yohimbine (5 x 10(-7) mol). These results suggest that phentolamine prevented brain-mediated hepatic glycogenolysis by a mechanism that may not be classified pharmacologically as involving either alpha 1- or alpha 2-receptors.     

Aging progressively decreases the densities and alters the diurnal rhythms of alpha 1-adrenergic receptors in selected hypothalamic regions

Associated with the decline in reproductive function that occurs with age in female rats are a diminished ability to secrete LH in response to ovariectomy as well as alterations in the preovulatory LH surge mechanisms. The noradrenergic system, an important regulator of LH release, exhibits age-related alterations in the pattern of neurotransmitter release. In addition, the density of alpha 1-receptors changes in intact old rats. To determine whether age-related changes in the pulsatile pattern of LH secretion are associated with declines in the densities and/or a loss of diurnal rhythm of alpha 1-receptors, we measured the density of alpha 1-receptors at various times of day in young, middle-aged, and old ovariectomized rats. By middle-age, mean concentrations of alpha 1-receptors decline in the median eminence and suprachiasmatic nucleus. By old age, mean concentrations of alpha 1-receptors decline in all regions of the hypothalamus examined but not in the dorsal lateral septum. In addition, the diurnal rhythms in alpha 1-receptors, which occur in the medial preoptic nucleus and the suprachiasmatic nucleus of young rats, are lost by middle-age. These data indicate that progressive alterations in the mean concentrations and diurnal rhythms of alpha 1-receptors occur with age in brain regions important for the regulation of reproductive functions and may contribute to the age-associated deficits in LH secretion.     

In the ventral tegmental area, progestins' membrane-mediated actions for lordosis of hamsters and rats involve protein kinase A

Progestin-facilitated lordosis of hamsters and rats is enhanced by activation of dopamine type 1 (D1) or GABAA/benzodiazepine receptor complexes (GBRs) in the ventral tegmental area (VTA) and these effects involve G-proteins and second messengers, such as adenosine 3',5'-monophosphate (cAMP). We examined whether D1- and/or GBR-mediated increases in progestin-facilitated lordosis of female hamsters and rats involve the cAMP-dependent protein kinase, protein kinase A (PKA), in the VTA. In experiment 1, ovariectomized hamsters, primed with estradiol (E2; 10 microg at h 0) + progesterone (P; 100 microg at h 45), were first pre-tested for lordosis and motor behavior (h 48) and then infused with the PKA inhibitor, Rp-cAMP (100 ng/side), or vehicle. Thirty minutes later, hamsters were retested and then received infusions of the D1 agonist, SKF38393 (100 ng/side), the GBR agonist, muscimol (100 ng/side), or vehicle to the VTA. Hamsters were post-tested for lordosis and motor behavior 30 min later. In Experiment 2, ovariectomized rats, primed with E2 (10 microg at h 0), were first pre-tested for lordosis and then infused with Rp-cAMP (100 ng/side) or vehicle to the VTA at h 44. Immediately after testing, rats received infusions of SKF38393 (100 ng/side), muscimol (100 ng/side), or vehicle and were retested for lordosis. Rats were then infused with the neurosteroid, 5alpha-pregnan-3alpha-ol-20-one (3alpha,5alpha-THP; 100 or 200 ng/side), or beta-cyclodextrin vehicle and were post-tested for lordosis and motor behavior 10 and 60 min later. The enhancing effects of progestins or progestins plus D1 or GBR activation on lordosis of E2-primed hamsters and rats were blocked by the PKA inhibitor, Rp-cAMP. Thus, in the VTA, progestins' membrane actions involving D1 or GBRs are mediated, in part, by PKA.     

Antagonism of sexual behavior in female rats by ventromedial hypothalamic implants of antiestrogen

The present experiments sought to identify brain regions in which implants of an antiestrogen would antagonize the ability of a systemic estradiol treatment to activate sexual behavior in female rats. In experiment 1, ovariectomized female rats were implanted subcutaneously with 5-mm Silastic capsules containing a 5% concentration of estradiol and injected with 500 micrograms progesterone 2 days later, 4-5 h before testing for sexual behavior. Bilateral intracranial implants of 1% crystalline concentrations of the high-affinity antiestrogens monohydroxytamoxifen (TAM) or keoxifene placed into the ventromedial nucleus of the hypothalamus (VM) 24 h prior to estradiol treatment significantly reduced lordosis responsivity compared with control females receiving empty cannulae. Implants of 1% TAM into the medial preoptic area or medial amygdala 24 h prior to estradiol that no significant effect on lordosis. Similarly, implants of 1% TAM into the VM 12 h after estradiol had no effect on lordosis. In experiment 2, lordosis was activated by subcutaneous implants of Silastic capsules containing 1% estradiol plus 500 micrograms progesterone. In this experiment, implants of 1% TAM into the VM 24 h prior to estradiol significantly reduced lordosis only if both cannulae tips were in, or adjacent to, the VM. Females receiving intracranial 1% TAM, but whose cannulae (even unilaterally) were outside the VM, had levels of lordosis equivalent to those of control females. Increasing the concentration of intracranial TAM to 10% virtually eliminated lordosis in females with bilateral implants in the VM, whereas females receiving intracranial 10% TAM in the region of, but outside, the VM showed no evidence of a lordosis deficit.(ABSTRACT TRUNCATED AT 250 WORDS)     

The excitation and inhibition of sexual receptivity in female hamsters by progesterone: time and dose relationships, neural localization and mechanisms of action.

Ovariectomized hamsters were administered estradiol benzoate (EB) and 44 h later, progesterone (P.) Lordosis behavior was induced. When an additional dose of P was given up to 24 h prior to or 24 h after the EB, EB-P facilitation of lordosis was inhibited. Additional hamsters were given varying doses of P (25-200 mu) following EB using both excitatory and inhibitory paradigms. Inhibition of EB-induced lordosis was effected with a lower dose of P than was the facilitation of EB induced lordosis by P. Hamsters were also given intracerebral implants of P using excitatory and inhibitory paradigms. No excitatory loci were found. Inhibition of EB-induced lordosis was effected by implants in the posterior hypothalamus and anterior mesencephalon, but not by diencephalic implants. Other hamsters were administered tritiated estradiol (E2) plus P prior to, concurrent with, or shortly after the E2. P had no effect upon the accumulation of E2 by any brain sites, although E2 was found to concentrate to a greater degree in the diencephalon than in the mesencephalon or cortex. The estrogen-induced depletion and replenishment of hypothalamic cytosol estrogen receptors was also studied. Concurrent P treatment had no effect upon the receptor depletion-replenishment process. It was concluded that P can both facilitate and inhibit estrogen-induced lordosis and that the inhibitory effects of P are not upon estrogen-sensitive cells in the brain.     

Progesterone enhancement of lysergic acid diethylamide and levo-5-hydroxytryptophan stimulation of the copulatory response in the female rat

Copulatory behavior in the ovariectomized rat, i.e. the lordosis response (LR) on being mounted by a male, can be induced by administration of either estrogen alone or estrogen followed by progesterone. LR has been shown to be inhibited by lysergic acid diethylamide (LSD) in certain doses (greater than or equal to 50 micrograms/kg) and by Levo-5-hydroxytryptophan (L-5-HTP) (greater than or equal to 2.5 mg/kg). This effect was recently found to be enhanced by increasing doses of progesterone. In contrast, small doses of LSD (5-30 micrograms/kg) have been shown to increase LR activated by estrogen alone. The effects of various hormone treatments on the stimulatory action of LSD were tested in the present study. When the lordosis behavior was activated by estradiol benzoate (EB) alone (25 or 7 X 2 micrograms/kg), LR increased 10 min after injection of LSD in a dose of 10 micrograms/kg. There were no detectable effects in animals treated with estrogen alone when the dose of LSD was lowered to 1 microgram/kg. LSD in the latter dose gave an increased response, however, when LR was activated by EB (5 micrograms/kg) in combination with progesterone (4.0 mg/rat). An analogous study was conducted with L-5-HTP, after pretreatment with pargyline and R04-4602. Small doses of L-5-HTP (0.25 and 0.05 mg/kg) stimulated the LR and the influence of progesterone was the same as for small doses of LSD. Possible mechanisms underlying the observed influence of progesterone on serotonergic mechanisms involved in the lordosis behavior are discussed.