Effects of intracerebroventricular leptin administration on feeding and sexual behaviors in lean and obese female Zucker rats.

Obese Zucker rats (fa/fa) are characterized by inadequate leptin signaling caused by a mutation in the leptin receptor gene. Obese Zucker females are infertile and hyporesponsive to the inductive effects of ovarian hormones on sexual behaviors. Leptin treatment reverses aspects of reproductive dysfunction due to perturbations in energy balance in other animal models. Our first experiment tested the hypothesis that intracerebroventricular (icv) leptin administration would enhance the display of sexual behaviors in obese Zucker females. A second experiment compared lean and obese Zucker females' responses to leptin, during fed and fasted conditions. Ovariectomized (OVX) Zucker rats were implanted with lateral ventricular cannulae. In Experiment 1, fasted, obese females received estradiol benzoate, progesterone, and icv injections of 3, 18, or 36 microg murine leptin or vehicle. Leptin administration reduced food intake, but did not enhance sexual behaviors. In Experiment 2, steroid-replaced, OVX lean and obese females (from a different source than those in Experiment 1) received icv injections of vehicle or 3 or 36 microg leptin under fed and fasted conditions. Leptin treatment reduced food intake and weight gain in the fed, but not the fasted, condition in both genotypes. Sexual receptivity and locomotion were not affected, but icv leptin injections reduced proceptive behaviors in ad libitum-fed rats. These data confirm previous reports that centrally administered leptin decreases food intake and weight gain in obese Zucker rats; results from Experiment 2 suggest that lean and obese females are similarly responsive to these actions of leptin. Contrary to our hypothesis, leptin treatment did not stimulate sexual behaviors; rather, the hormone appears to inhibit the display of sexual proceptivity in ad libitum-fed lean and obese Zucker female rats.

Deficits in progesterone-facilitated sexual behaviors and forebrain estrogen and progestin receptors in obese female Zucker rats.

Obese female Zucker rats (fa/fa) are sterile. Among their reproductive abnormalities is hyporesponsiveness to the stimulatory effects of ovarian steroid hormones on sexual behaviors. This study was designed to test the hypothesis that obese Zucker females are deficient in hypothalamic/preoptic area estrogen receptors (ERs) and/or estradiol-induced progestin receptors (PRs). Ovariectomized (OVX) lean and obese Zucker rats were tested for the display of sexual behaviors following injection of estradiol benzoate (EB, 15 or 100 microg/kg) plus progesterone (P, 2 mg/kg). As expected, obese females showed significantly lower lordosis quotients and lordosis ratings than lean animals after injection of the lower, physiological dose of EB followed by P. In contrast, obese and lean females receiving the higher EB dose, prior to P, showed similar levels of sexual receptivity. Two weeks later, these OVX lean and obese females received injections of vehicle, 15 or 100 microg/kg EB, prior to perfusion and tissue processing for PR immunocytochemistry (ICC). Additional groups of OVX virgin females of both genotypes were perfused and tissue from the preoptic area and hypothalamus was processed for ER alpha ICC. No genotypic differences in the number of cells containing ER alpha-immunoreactivity (-IR) in the medial preoptic area (MPOA), ventromedial hypothalamus (VMH) or arcuate nucleus (ARC) were noted, but obese females had significantly fewer ER alpha-IR cells in the anteroventral periventricular nucleus (AVPV) than lean rats. In both genotypes, the number of PR-IR cells in the AVPV, MPOA and VMH was significantly higher following injection of EB (either dose) as compared to vehicle, demonstrating estradiol induction of PRs. Only in the MPOA was there a significant difference between fat and lean females in estradiol-induced PR-IR. Obese females receiving 15 microg/kg EB had fewer PR-IR cells in the MPOA than comparably-treated lean animals. No such genotypic difference was observed following injection of the vehicle or higher dose (100 microg/kg) of EB. These data are consistent with the hypothesis that deficiencies in ER alpha in the AVPV and/or PRs in the MPOA may contribute to obese Zucker females' poor responsiveness to ovarian steroid hormones.

Sexual motivation in the male rat: the role of primary incentives and copulatory experience.

Two experiments explored the motivational impact of primary incentive female cues on the operant behavior of sexually naive and experienced male rats. In the first experiment, a straight-arm runway was used to assess the subjects' motivation to approach a goalbox containing either male or female "targets." Twelve sexually naive Long-Evans males ran for: (1) an empty goalbox; (2) a male conspecific; (3) an ovariectomized (OVX) female; (4) an OVX female given estradiol; (5) or an OVX female treated with estradiol and progesterone. A perforated Plexiglas partition in the goalbox prevented the subject males from physically interacting with the targets, although olfactory, visual, and auditory cues were accessible. We hypothesized that subjects would manifest shorter run times (reflecting greater motivation) when the goalbox contained a receptive/proceptive female as opposed to a nonreceptive female target. Subjects' run times were ordered depending on the nature of the target (from slowest to fastest): empty goalbox, male conspecific, OVX female, OVX + estradiol female, and OVX + estradiol + progesterone female. As predicted, subjects ran significantly faster for a receptive/proceptive female than for a nonreceptive female, indicating that sexually naive males are inherently motivated by female precopulatory cues. In the second experiment, 30 sexually naive male subjects ran for a goalbox containing either a nonestrous (OVX) or an estrous (OVX + estradiol + progesterone) female. Following six trials, 10 males were allowed one intromission with a receptive female, 10 males experienced one ejaculation, and 10 remained sexually naive. Only those males having experienced an ejaculation subsequently decreased their run times for both nonestrous and estrous females, indicating that sexual reinforcement produced by ejaculation, but not intromission, further enhances the motivational impact of female incentive cues.

Opioid receptor blockade promotes weight loss and improves the display of sexual behaviors in obese Zucker female rats.

Obese Zucker female rats are hyperphagic, overweight, infertile, and hyporesponsive to the inductive effects of ovarian steroid hormones on sexual behaviors. It has been postulated that endogenous opioid activity may contribute to their obesity and reproductive dysfunction. To test this hypothesis, ovariectomized, adult obese Zucker rats were treated with the opioid receptor antagonist, naltrexone, or saline prior to measurement of steroid-induced sexual behaviors, food intake, and body weight. In estradiol benzoate (EB)-treated rats, naltrexone injection increased the display of sexual receptivity (lordosis quotient, LQ: saline, 11+/-10%; 5 mg/kg naltrexone, 54+/-15%, p < 0.05) and also elicited proceptivity (PRO), which was never observed after saline injection. In EB plus progesterone-treated animals, naltrexone administration enhanced both sexual receptivity and proceptivity (LQ: saline, 17+/-10%; 5 mg/kg naltrexone, 96+/-3%; p < 0.05; PRO: saline, 3.0+/-2.4 bouts/min; 5 mg/kg naltrexone, 45.3+/-12 bouts/min; p < 0.01). Naltrexone injection also decreased 24-h food intake (saline, 24.2+/-0.7 g; 5 mg/kg naltrexone, 17.6+/-1.2 g; p < 0.05) and weight change (saline, +7.3+/-0.8 g; 5 mg/kg naltrexone, -4.5+/-1.4 g, p < 0.01). Morphine treatment blocked these effects of naltrexone on sexual behaviors, food intake, and body weight. These data suggest that endogenous opioids contribute to hyperphagia, obesity, and behavioral hyporesponsiveness to ovarian steroid hormones in obese Zucker rats.

Food restriction neither improves nor exacerbates reproduction in obese female Zucker rats.

Obese female Zucker rats have several reproductive abnormalities, including delayed puberty, abnormal estrous cyclicity, and behavioral hyporesponsiveness to ovarian steroid hormones. To ascertain whether excessive body weight per se causes these reproductive abnormalities, obese Zucker female rats were fed ad lib or were food restricted to match their body weights to those of lean counterparts. Food restriction neither accelerated vaginal opening nor normalized estrous cyclicity in obese female rats. Following ovariectomy, an injection of estradiol benzoate (EB, 15 microg/kg, s.c.) induced extremely low sexual receptivity in all rats, and proceptive behaviors were never observed. After treatment with EB plus progesterone (P, 2 mg/kg, s.c.), lean rats were very receptive (lordosis quotient, LQ = 94 +/- 2%) and proceptive (PRO = 12.5 +/- 2 events/min) while both ad lib-fed and food-restricted obese rats were only marginally receptive and proceptive (LQ= 19 +/- 9%, PRO = 1.8 +/- 0.7 events/min; LQ = 31 +/- 15%, PRO = 4.7 +/- 3 events/min, respectively). A higher progesterone dose (20 mg/kg) elicited vigorous sexual receptivity (LQ = 88-99%) and proceptivity (PRO = 16.5-20.4 events/min) in all EB-treated rats. Adiposity was significantly lower in food-restricted obese rats as compared to ad lib-fed obese rats (36.5 +/- 1.7% vs. 69.4 +/- 2.7%), but greater than that observed in lean rats (24.4 +/- 1.1%). These data suggest that excessive body weight per se does not underlie reproductive abnormalities in obese Zucker rats, but do not rule out the possibility that excessive adiposity may contribute to their infertility.

Intraventricular injection of neuropeptide Y antisera curbs weight gain and feeding, and increases the display of sexual behaviors in obese Zucker female rats.

Obese Zucker rats are hyperphagic, overweight, and infertile. It has been postulated that neuropeptide Y (NPY) overproduction may contribute to obesity and infertility in these animals. To test this hypothesis, ovariectomized, adult obese Zucker rats were implanted with cannulae in the third ventricle and subsequently injected with NPY antisera or normal rabbit sera (NRS) 6, 4 and 2 h before experimental observation. Steroid-treated females injected with NPY antisera were significantly more receptive and were more likely to show proceptive behaviors than after treatment with NRS (e.g., lordosis quotient: NPY antisera, 65.5+/-6.9%; NRS, 30.9+/-11.6%, P < 0.02; 91% displaying proceptivity after NPY antisera injection vs. 36% after NRS, P < 0.03). Injection of NPY antisera also curbed food intake and weight gain (24 h food intake: NPY antisera, 10.5+/-2.1 g; NRS, 20.5+/-1.7 g, P < 0.01; 24 h weight gain: NPY antisera, -5.4+/-2.2 g; NRS, 5.8+/-0.7 g, P < 0.01). Locomotor activity was similar after NRS and NPY antisera treatment (P > 0.5) suggesting that general malaise was not responsible for the effects of NPY antisera on food intake or body weight. These data suggest that endogenous neuropeptide Y contributes to excessive feeding and weight gain, and suppressed reproductive behaviors in obese Zucker female rats.

Site-specific opioid receptor blockade allows prepubertal guinea pigs to display progesterone-facilitated lordosis.

Ovariectomized (OVX) juvenile guinea pigs (approximately 3 weeks old) rarely display steroid-induced sexual receptivity. Systemic administration of the opioid receptor antagonist naloxone enhances the display of progesterone-facilitated lordosis in prepubertal females, suggesting that endogenous opioids tonically inhibit the expression of sexual receptivity at this age. This study was designed to ascertain the neural site(s) at which naloxone injection would stimulate lordosis in juvenile guinea pigs. Hartley guinea pigs were OVX at 10-11 days of age and 2-6 days later implanted with bilateral cannulae aimed at the medial preoptic area/anterior hypothalamus (MPOA/AH), ventrolateral hypothalamus/ventromedial hypothalamus (VLH/VMH), or mesencephalic central gray (MCG). At 21-23 days of age, following administration of estradiol benzoate (10 microgram(s)) and progesterone (0.5 mg), naloxone (100 ng/side) or 0.9% saline was injected through the cannulae and the guinea pigs were tested for the display of lordosis. The MPOA/AH was the only site at which application of naloxone reliably elicited lordosis (87% positive response vs 12% for saline). Few females (< 17%) displayed lordosis following injections of naloxone or saline into the VLH/VMH or MCG. A second experiment demonstrated that the stimulation of lordosis following MPOA/AH naloxone application was prevented by prior injection of the opioid agonist morphine (500 ng/side) at the same site. These data support the hypothesis that endogenous opioids acting in the MPOA/AH, but not the VLH/VMH or MCG, tonically inhibit the display of progesterone-facilitated lordosis in prepubertal guinea pigs.

Lordosis-enhancing medial preoptic area lesions do not alter hypothalamic estrogen receptor- or progestin receptor-immunoreactivity in prepubertal female guinea pigs.

Female guinea pigs rarely display adult-typical lordosis responses to ovarian steroid hormones until 40-50 days of age. Behavioral hyporesponsiveness in prepubertal females may be due, in part, to deficiencies in hypothalamic estrogen receptors and/or estradiol-induced progestin receptors. This study was designed to test the hypothesis that bilateral medial preoptic area (MPOA) lesions, which enhance the display of progesterone-facilitated lordosis in juvenile females, increase levels of hypothalamic estrogen receptors and/or estradiol-induced progestin receptors. Hartley guinea pigs were ovariectomized at 11-12 days of age and at 14-15 days of age received bilateral electrolytic or sham lesions aimed at the MPOA. At approximately 3 weeks of age, lesioned and sham-lesioned animals were either tested for the display of progesterone-facilitated lordosis or perfused, and their hypothalamic tissue processed for estrogen receptor- or estradiol-induced progestin receptor-immunostaining. Although a significantly higher percentage of MPOA-lesioned than sham-lesioned guinea pigs displayed progesterone-facilitated lordosis (85.7% vs. 5. 8%, respectively, p<0.05), there were no significant lesion-related differences in the number or staining intensity of cells containing estrogen receptor- or estradiol-induced progestin receptor-immunoreactivity in the ventrolateral hypothalamus or arcuate nucleus. These data do not support the hypothesis that the enhanced display of progesterone-facilitated lordosis in prepubertal guinea pigs following MPOA lesions is due to increased hypothalamic concentrations of estrogen receptors or estradiol-induced progestin receptors.

Reproductive behavioral responsiveness to noradrenergic stimulation in developing guinea pigs.

The stimulatory effects of ovarian hormones on sexual receptivity in guinea pigs may be mediated by norepinephrine. Juvenile females rarely exhibit steroid-induced receptivity and also respond poorly to the lordosis-enhancing action of alpha-noradrenergic receptor stimulation. This experiment was designed to chart the development of behavioral responsiveness to the alpha-noradrenergic agonist, clonidine, and to test the hypothesis that higher doses of estradiol and/or clonidine are required to stimulate lordosis in juvenile compared to adult guinea pigs. Ovariectomized females received estradiol benzoate (10 microg or 50 microg SC) 40-48 h before administration of clonidine (1 mg/kg or 5 mg/ kg IP) or saline at 14-17-day intervals. Regardless of treatment, few animals (0-36%) displayed lordosis at 20, 34, or 48 days of age. At 65 days of age, in both estradiol dose groups significantly more clonidine- (1 mg/kg) than saline-injected animals displayed lordosis (80-91 vs. 0-33%, respectively). Clonidine (5 mg/kg) was ineffective at all ages. These data do not support the hypothesis that behavioral responsiveness to alpha-noradrenergic receptor stimulation in immature females can be elicited by increasing the doses of estradiol and/or clonidine. These results suggest the occurrence of a maturational change in the neural systems governing noradrenergic involvement in steroid-induced sexual behavior in guinea pigs.

Effects of medial preoptic area and septal lesions on puberty in female guinea pigs.

Lesions of the medial preoptic area (MPOA) result in the robust display of steroid-induced lordosis by otherwise unresponsive, immature guinea pigs. This study was designed to test the hypothesis that destruction of this region also hastens the onset of ovarian cyclicity, and to ascertain whether lesions of the septum produce effects on puberty similar to those observed following MPOA lesions. Juvenile (13-15 days old), ovariectomized or ovary-intact guinea pigs received bilateral electrolytic or sham lesions of the MPOA or septum. Bilateral lesions of the septum did not influence the display of lordosis or any aspect of estrous cyclicity. However, a higher percentage of guinea pigs with MPOA lesions than with sham lesions displayed progesterone-facilitated lordosis at 22, 29, and 39 days of age; by 49 days of age, the two groups responded in a similar, robust fashion. Females with MPOA lesions were significantly heavier and older than sham-lesioned or nonmanipulated females on the day of first vaginal opening. These data suggest that activity in the MPOA tonically inhibits the display of progesterone-facilitated lordosis through at least 39 days of age but does not suppress the onset of ovarian cyclicity in guinea pigs. Contrary to our hypothesis and in contrast to what has been observed in rats, signals originating in or passing through the MPOA may be required in order for estrous cyclicity to be initiated at the appropriate age. Furthermore, neither of these effects of MPOA lesions appears to be due to damaging efferent or afferent projections through the septum.

Abnormal estrous cyclicity and behavioral hyporesponsiveness to ovarian hormones in genetically obese Zucker female rats.

Obese Zucker female rats are infertile. The present study was designed to assess estrous cyclicity in adult, ovary-intact, lean and obese Zucker rats and to compare reproductive behaviors induced by exogenous steroid hormones in ovariectomized (ovx) lean and obese Zucker rats. The majority (90%) of obese rats had incomplete cycles in comparison with the normal, 4-day cycles displayed by lean Zucker rats. After ovariectomy, all rats were treated with estradiol benzoate (EB, 15-100 micrograms/kg) or EB plus progesterone (P, 2-20 mg/kg), and tested for sexual receptivity and proceptivity (PRO). At the highest EB dose, obese Zucker females displayed lordosis less frequently than lean rats (lordosis quotient, LQ, 8 +/- 6% vs. 32 +/- 13%, respectively). At the lowest doses of EB plus P, lean females were extremely receptive and proceptive (LQ = 93 +/- 4%, PRO = 6.2 +/- 2 bouts/min). Zucker obese females, in contrast, were only slightly receptive (LQ = 26 +/- 11%) and showed less PRO than lean rats (PRO = 2.4 +/- 0.6 bouts/min). Increasing the dose of either EB or P, administered in combination with the lowest dose of the other hormone, produced receptivity and PRO in obese Zucker females that were comparable with those observed in lean rats. Serum estradiol and P levels in ovx obese rats were either equivalent to or higher than those in the ovx lean rats when both were given the same doses of hormones. These data suggest that considerably higher doses and serum concentrations of EB and/or P are required to elicit robust lordosis and PRO in ovx obese Zucker than in lean rats. This behavioral hyporesponsiveness to sex steroid hormones may contribute to infertility in the obese Zucker female rat.

Maternal insulin manipulations in rats organize body weight and noradrenergic innervation of the hypothalamus in gonadally intact male offspring.

In previous work it has been shown that adult male, but not female, offspring of rats that have been injected with protamine zinc insulin (6 IU/kg) on days 15-20 of gestation, develop significant obesity beginning about 50 days of age. This obesity is accompanied by elevated medial hypothalamic extracellular norepinephrine levels. To examine whether the expression of obesity in male offspring is mediated by perinatal testosterone levels, male offspring of insulin-treated or control dams were either castrated or received sham surgery on postnatal day 1. Castrated male offspring of insulin-treated dams did not become obese like their gonadally intact male littermates. This suggests that perinatal testosterone levels may interact with developmental processes mediating the obesity in male offspring of insulin-treated dams. A second question addressed was whether the elevated hypothalamic extracellular norepinephrine levels observed in our earlier work are evident as morphological changes in norepinephrine-containing systems in the medial hypothalamus and locus coeruleus. We found a significant enhancement of dopamine-beta-hydroxylase immunoreactivity in fibers innervating the paraventricular nucleus of the hypothalamus in 121-day-old, gonadally intact male offspring of insulin-injected dams. This suggests that the impact of maternal insulin injections on offspring obesity may be mediated through its organizing action on feeding-related fibers in the paraventricular nucleus.

Progesterone-facilitated lordosis in medial preoptic area-lesioned, juvenile guinea pigs.

Juvenile female guinea pigs rarely display lordosis in response to estradiol and progesterone treatments that elicit sexual receptivity in adults. To test the hypothesis that the medial preoptic area (MPOA) tonically inhibits the display of steroid-induced lordosis in juveniles, 11-day-old guinea pigs were ovariectomized (OVX) and received bilateral, sham, or electrolytic lesions aimed at the MPOA 3-4 days later. At 20-22 days of age, these females were tested for the expression of sexual receptivity following injections of estradiol benzoate (EB, 10 micrograms s.c.) and progesterone (0.5 mg s.c., 40 h after EB). The lesions damaged portions of the MPOA, the nucleus of the diagonal band of Broca, the lateral aspect of the medial preoptic nucleus, the medial part of the preventricular portion of the periventricular nucleus, and the anterior commissure. The lesions did not alter the display of estradiol-induced lordosis. However, after treatment with EB plus progesterone, 20% of the sham-lesioned females displayed lordosis, as compared to 80% of the MPOA-lesioned animals. These data are consistent with the hypothesis that neurons originating in and/or traversing the MPOA tonically suppress the display of progesterone-facilitated lordosis in juvenile guinea pigs. Removal of this inhibitory input allows prepubertal females to respond behaviorally to estradiol and progesterone in an adult-typical fashion.

Maternal hormonal manipulations in rats cause obesity and increase medial hypothalamic norepinephrine release in male offspring.

In previous work it has been shown that adult male, but not female, offspring of rats that have either been injected with Protamine Zinc Insulin on days 15-20 gestation, or undernourished during the first 2 weeks of gestation, develop significant obesity commencing at about 50 days of age. The present experiment examines the question of whether rats with these two forms of obesity display neurochemical abnormalities in areas of the brain known to influence food intake and body weight. Twenty-one gauge stainless steel guide shafts were surgically implanted using standard stereotaxic procedures. One week later 26 ga microdialysis probes were lowered into the medial hypothalamus. Dialysates collected from male offspring in the two experimental conditions contain significantly higher norepinephrine (NE) levels than did controls. It would appear that in addition to sharing a similar time course of onset and a sex dependent expression of obesity, both of these models are also characterized by elevated medial hypothalamic NE. Since this obesity appears only in males, and at a time when testosterone levels are rapidly rising in males, and since testosterone has been shown to elevate food intake and body weights in rats, we also investigated whether gonadal weights or circulating testosterone levels were differentially elevated by our manipulations.

Hypothalamic estrogen receptor-immunoreactivity in prepubertal vs adult female guinea pigs.

Juvenile guinea pigs (18-20 days old) rarely display lordosis in response to estradiol and progesterone treatments that elicit sexual behavior in adult females. Nor do immature animals release a preovulatory-like surge of luteinizing hormone in response to estradiol. In vitro radioligand binding assays have revealed similar concentrations of estrogen receptors in the hypothalamus and preoptic area of prepubertal and adult guinea pigs. The aim of the present study was to compare estrogen receptor-immunoreactivity in a variety of forebrain regions of immature and adult guinea pigs, to determine whether age differences in estrogen receptor levels in more discrete portions of the hypothalamus and preoptic area exist. Forebrain tissue from juvenile (17 days) and adult females (> 6 weeks), ovariectomized 6 days previously, was processed for estrogen receptor-immunoreactivity, using Abbott Laboratories' H222 anti-human estrogen receptor antibody. Juveniles had estrogen receptor-immunoreactive cells in all of the same regions as adults: medial preoptic area, medial preoptic nucleus, bed nucleus of the stria terminalis, periventricular, paraventricular, dorsomedial and arcuate nuclei, ventrolateral and anterior hypothalamic regions, and amygdala. Among the areas in which estrogen receptor-immunoreactivity was quantified (medial preoptic area, medial preoptic nucleus, anterior periventricular nucleus, arcuate nucleus and ventrolateral hypothalamus), the only region in which an age difference in estrogen receptor-immunostaining was observed was the rostral portion of the ventrolateral hypothalamus. Juvenile females had, on average, 30% fewer estrogen receptor-immunoreactive cells in a sample of this region than adults (440 +/- 25 vs. 626 +/- 25, P = 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of alcohol on the sexual motivation of the male rat.

Previous research measuring the effects of alcohol on sexual behavior has primarily focused on its effects on copulation. The present experiment was designed to investigate the effects of alcohol on the sexual motivation of the male rat by requiring operant responding to gain access to a sexually receptive female. A novel apparatus was used that allowed both visual and olfactory cues from an estrous female to reach the male. Lever presses resulted in the opening of a door that permitted the male rat to enter an adjacent chamber where a receptive female was located. Treatment with low to moderate doses of alcohol (0.5 g/kg and 1.0 g/kg) resulted in increased latencies to emit the first response of the males working for access to females, but did not affect response rate or subsequent mount or ejaculation latencies, when these males were allowed access to the receptive female. Furthermore, alcohol failed to show any response-reinstating or disinhibitory effects when tested following a period of nonreinforced extinction trials. An additional experiment, in which rats received equivalent doses of alcohol, revealed no decrease in spontaneous locomotion. Taken together, these data suggest that the response-reducing effects of alcohol are probably not a result of general drug-induced reductions in activity, but rather an attenuating action of the drug on sexual motivation.

Opiate receptor blockade enhances the display of progesterone-facilitated lordosis in juvenile female guinea pigs.

Juvenile female guinea pigs rarely display estradiol-induced or progesterone-facilitated sexual receptivity. This experiment was designed to test the hypothesis that endogenous opiate peptides tonically inhibit the display of steroid-induced lordosis in juvenile females. Hartley guinea pigs were ovariectomized (OVX) at 10-11 days of age and treated with estradiol benzoate (EB, 10 micrograms) 5-7 days later. Injection of the opiate antagonist, naloxone (2.0 mg/kg), did not facilitate lordosis (compared to vehicle) in EB-primed, OVX juveniles; when these animals were subsequently given progesterone, and the treatment groups were reversed (i.e., the group previously treated with vehicle received 2.0 mg/kg naloxone 4.5 hr after progesterone, and vice versa), four of six naloxone-injected animals displayed lordosis, compared to one of six control animals. In a second experiment, a range of doses of naloxone was administered to EB-plus progesterone-treated OVX juveniles. Naloxone at the 2.0 mg/kg dose resulted in a significant increase in the percentage of females displaying lordosis, with lower doses (0.5-1.0 mg/kg) producing responses intermediate between those observed in animals receiving the 2.0 mg/kg dose and the vehicle. Furthermore, the facilitatory effect of naloxone on sexual receptivity was completely blocked by concomitant injection of morphine (10.0 mg/kg). These data suggest that endogenous opiates tonically inhibit the display of progesterone-facilitated lordosis in juvenile female guinea pigs.

Ibotenic acid-induced lesions of the medial preoptic area/anterior hypothalamus enhance the display of progesterone-facilitated lordosis in male rats.

Electrical lesions of the medial preoptic area/anterior hypothalamus (MPOA/AH) have been reported to enhance the display of steroid-induced lordosis in castrated male rats. This study employed the cell body-specific neurotoxin, ibotenic acid, to ascertain whether neurons originating in this region (as opposed to axons of passage) tonically inhibit steroid-induced lordosis in adult male rats. Castrated, adult Long-Evans males received bilateral electrical lesions or injections of ibotenic acid or vehicle aimed at the MPOA/AH. Following administration of estradiol benzoate (EB) and progesterone, lordosis quotients (LQs) and lordosis ratings (LRs) were significantly higher in groups of rats with electrical lesions (LQ = 62.2 +/- 15.1; LR = 1.22 +/- 0.34) and ibotenic acid-induced lesions (LQ = 58.1 +/- 12.2; LR = 0.99 +/- 0.24) than in the control group (LQ = 12.8 +/- 7.3; LR = 0.22 +/- 0.13). To determine whether this enhancement of receptive behavior in MPOA/AH-lesioned males was an effect on estradiol-induced, as compared to progesterone-facilitated lordosis, groups of castrated rats in a second experiment received bilateral injections of ibotenic acid or vehicle aimed at the MPOA/AH and were tested for lordosis after administration of EB alone and again after injection of progesterone. Following treatment with EB alone, rats with ibotenic acid-induced MPOA/AH lesions tended to be slightly less receptive than control animals. However, following injections of progesterone, LQs and LRs were higher in the MPOA/AH-lesioned group than in the control animals, as had been observed in the first experiment.(ABSTRACT TRUNCATED AT 250 WORDS)

Colchicine-induced accumulation of estrogen receptor and progestin receptor immunoreactivity in atypical areas in guinea-pig brain.

Using immunocytochemical techniques, cells containing estrogen and progestin receptors have been observed in many discrete regions of the guinea-pig forebrain, including the mediobasal hypothalamus and preoptic area. While most reaction product is located within cell nuclei, we have reported abundant reaction product in perikaryal cytoplasm and neuronal processes as well. Ultrastructural analysis has revealed the presence of estrogen and progestin receptors in atypical subcellular sites within the hypothalamus, including dendrites and axon terminals. In order to determine if microtubule-dependent intracellular transport is involved in intraneuronal transport of steroid hormone receptors, ovariectomized guinea-pigs were injected intracerebroventricularly with the microtubule inhibitor, colchicine, and brain sections at the level of the hypothalamus were immunostained for estrogen receptors. This treatment resulted in the appearance of estrogen receptor immunoreactivity in the paraventricular and mediodorsal thalamic region, areas typically devoid of estrogen receptor-immunoreactive cells in guinea-pigs. In a second study on progestin receptors, we observed the colchicine-induced accumulation of progestin receptor immunoreactivity in the paraventricular thalamic, mediodorsal thalamic and lateral dorsal thalamic areas as well as in the medial amygdala, all areas typically devoid of progestin receptor immunoreactivity. While estradiol injection induced progestin receptor immunoreactivity in the hypothalamus and preoptic area as described previously, it had no effect on the colchicine-induced accumulation in the thalamus and amygdala. These results provide evidence that in some neurons, progestin receptors and estrogen receptors are transported intracellularly, apparently at a rapid enough rate that they do not ordinarily accumulate within the perikaryon.

Estradiol pulses induce progestin receptors selectively in substance P-immunoreactive neurons in the ventrolateral hypothalamus of female guinea pigs.

Low doses of estradiol, administered as pulses, are as effective as higher doses for priming ovariectomized (OVX) guinea pigs to display progesterone-facilitated lordosis. High doses of estradiol, administered by constant-release implants, induce progestin receptors in many substance P-immunoreactive (SP-IR) neurons in the ventrolateral hypothalamus (VLH), a site at which estradiol primes OVX guinea pigs to respond behaviorally to progesterone. To test the hypothesis that behaviorally effective estradiol pulses induce progestin receptors selectively in substance P-containing neurons in the VLH, OVX females received estradiol implants 1 week prior to perfusion, or two pulses of estradiol-17 beta, injected 39 and 11 h before perfusion. Colchicine was administered intracerebroventricularly prior to perfusion. No significant differences were observed in the total number of progestin receptor-immunoreactive (PR-IR) or substance P-immunoreactive cells in the VLH and VLH/ventromedial hypothalamus (VMH), respectively, of females receiving the two estradiol treatments. However, the percentage of PR-IR cells in the VLH also immunoreactive for SP was significantly higher in the estradiol pulse-treated (53%), than in the estradiol capsule-implanted animals (36%). These data suggest that behaviorally effective estradiol pulses induce progestin receptors selectively in substance P-containing neurons in the VLH and are consistent with the hypothesis that substance P is involved in progesterone-facilitated lordosis in guinea pigs.